肿瘤领域文献阅读汇编

Advances presented at the 2026 American Association for Cancer Research (AACR) Annual Meeting highlight a shift from standalone artificial intelligence (AI) models to integrated, agentic systems across oncology. Platforms such as Synapse enable large-scale data coordination, supporting interoperable and reproducible research. Building on this foundation, conversational and multi-agent AI tools (e.g., DrBioRight, GP CoPilot, Isabl AI Agent) allow natural language interaction with multimodal cancer data, lowering technical barriers. Agentic frameworks for real-world data (RWD) transformation, including clinical document abstraction, cohort extraction, and social determinants of health (SDOH) analysis, demonstrate high accuracy and scalability, while self-critical systems improve reliability. Clinically, AI shows growing impact through validated imaging biomarkers, enhanced trial matching, and scalable cohort identification. In parallel, multi-agent systems are accelerating therapeutic discovery, including CAR-T development and immunotherapy target identification. Collectively, these advances position AI as an active, collaborative partner in cancer research and precision oncology.

Immune checkpoint inhibitors (ICIs) are a standard treatment across cancers, yet most patients do not respond, and existing biomarkers generalize poorly across tumor types and therapies. Here we present COMPASS, a pan-cancer foundation model that predicts immunotherapy response from bulk tumor transcriptomes using a concept bottleneck transformer. COMPASS encodes gene expression through 44 biologically grounded immune concepts representing immune cell states, tumor-microenvironment interaction and signaling pathways. Trained on 10,184 tumors across 33 cancer types, COMPASS achieves better average performance than 22 methods across 16 clinical cohorts spanning seven cancers and six ICIs, improving accuracy by 8.5% and area under the precision-recall curve by 15.7% on average across cohorts. COMPASS generalizes to cancer types and treatments not represented during fine-tuning and may inform indication selection and patient stratification. In survival analyses, patients classified by COMPASS as responders had longer overall survival (hazard ratio = 4.7, P < 0.0001). Personalized response maps connect gene expression to immune concepts, identifying programs associated with response and resistance; in immune-inflamed non-responders, COMPASS highlights programs including TGFβ signaling, endothelial exclusion, CD4+ T cell dysfunction and B cell deficiency. COMPASS predicts immunotherapy response and provides hypothesis-generating mechanistic insight for trial design and translational studies.

Fibroblast activation protein (FAP) is highly expressed in cancer-associated fibroblasts and represents an emerging target for radioligand therapy (RLT). This study aimed to evaluate treatment outcomes of ¹⁷⁷Lu-FAP-2286 and to investigate whether changes in quantitative parameters on ⁶⁸Ga-FAP-2286 PET/CT are associated with treatment response and survival in patients with advanced solid tumors. In this single-center retrospective study, patients with advanced solid tumors who completed 2 cycles of ¹⁷⁷Lu-FAP-2286 therapy and underwent both baseline and posttreatment ⁶⁸Ga-FAP-2286 PET/CT (8-16 wk after therapy) were included. Maximum standardized uptake value (SUVmax) and total tumor volume (TTV) were quantified using semiautomatic whole-body segmentation. Relative changes (ΔSUVmax and ΔTTV) were calculated. RECIST 1.1 served as the clinical reference standard. Receiver operating characteristic analysis assessed the ability of PET-derived changes to discriminate progressive disease. Progression-free survival (PFS) and overall survival (OS) were evaluated using Kaplan-Meier analysis and Cox proportional hazards models. Thirty patients (median age, 65 y; range, 23-78 y) completed 2 cycles of ¹⁷⁷Lu-FAP-2286 therapy. According to RECIST 1.1, partial response, stable disease, and progressive disease occurred in 6 (20.0%), 11 (36.7%), and 13 (43.3%) patients, respectively. SUVmax and TTV decreased in 46.7% of patients. ΔSUVmax demonstrated moderate discrimination for RECIST-defined PD (AUC 0.72, P=0.04). Patients with ΔSUVmax <30% had significantly longer PFS than those with ΔSUVmax ≥30% (median 10.3 vs. 4.4 mo; log-rank P<0.001). Median OS was not reached in the ΔSUVmax <30% group compared with 9.8 months in the ΔSUVmax ≥30% group (log-rank P=0.003). In multivariable analysis, ΔSUVmax remained independently associated with PFS (HR: 5.2, 95% CI 1.5-7.8; P=0.010) and OS (HR: 4.3, 95% CI: 1.3-14.1; P=0.015). In patients with advanced solid tumors treated with 2 cycles of ¹⁷⁷Lu-FAP-2286, posttreatment changes in SUVmax on ⁶⁸Ga-FAP-2286 PET/CT were associated with treatment response and survival, suggesting potential utility as an imaging biomarker for early response assessment and prognostic stratification.

Objectives. To evaluate the risks of mortality, cardiovascular events, and pulmonary complications among cancer patients who quit, continued smoking, or switched to e-cigarettes after diagnosis. Methods. Using the Korean National Health Insurance Service database (2015-2022), we identified 46 834 cancer patients who were smoking at diagnosis. Patients were categorized as continued smokers (n = 17 418), quitters (n = 25 909), or e-cigarette users (n = 3507) after diagnosis. We used multivariable Cox proportional hazards models to assess associations. Results. During a median 4.2 years of follow-up, all-cause mortality was significantly lower in quitters (hazard ratio [HR] = 0.92; 95% confidence interval [CI] = 0.86, 0.98) and e-cigarette users (HR = 0.84; 95% CI = 0.72, 0.99) than continued smokers. Both groups also showed lower cardiovascular event risks (HR = 0.64; 95% CI = 0.57, 0.72 and HR = 0.72; 95% CI = 0.53, 0.97, respectively). Compared with quitters, e-cigarette users had increased pulmonary complication risk (HR = 1.26; 95% CI = 1.00, 1.58). Conclusions. E-cigarette use was associated with lower mortality and cardiovascular risks than continued smoking, though complete cessation conferred greater benefits. Elevated pulmonary complications in e-cigarette users warrant caution. (Am J Public Health. Published online ahead of print July 2, 2026:e1-e12. https://doi.org/10.2105/AJPH.2026.308512).

Current sedentary behavior (SB) guidelines primarily emphasize total time spent sedentary. We explored differences between interrupted and prolonged SB in relation to a range of cancer outcomes. This study included 91,292 UK Biobank participants with valid accelerometer data. Participants were followed for a median of 12.38 years (interquartile range 11.56-13.15 years). A two-step approach based on a random forest model was used to classify SB. Multivariable Cox proportional hazards models were applied to overall incident cancers and cancer deaths, plus obesity-related and type-2 diabetes-related cancers, and 23 site-specific cancers. Models were adjusted for demographic, socioeconomic, lifestyle, dietary, and health-status factors, including age, ethnicity, deprivation, education, smoking, alcohol intake, diet, and morbidity count. Isotemporal substitution models were used to estimate the associated cancer risk when replacing prolonged SB with intermittent SB, or physical activity (PA). After adjusting for sociodemographic and lifestyle factors, each additional hour of prolonged SB was associated with a higher risk of overall cancer mortality (hazard ratio [HR] HR1hour 1.09; 95% confidence interval [CI] [1.06, 1.11]; p < 0.001). Replacing 1 hour per day of prolonged SB with light PA (HRLPA 0.88; 95% CI [0.79, 0.99]; p = 0.033) was associated with lower risk of overall cancer mortality. Similarly, replacing 30 min per day of prolonged SB with moderate PA (HRMPA 0.92; 95% CI [0.86, 0.99]; p = 0.024) was associated with a lower risk of overall cancer mortality. The main methodological limitations were observational design, residual confounding, healthy volunteer bias, and measurement imprecision due to having only 7 days of accelerometer wear. Cancer risk associated with SB is specific to prolonged SB. Replacing prolonged SB physical activity is associated with lower cancer risk.

In the US, Black patients with cancer consistently experience worse survival compared to White patients, even after adjusting for age, sex, and disease stage. Whether these disparities exist among patients receiving care in the Veterans Health Administration (VHA), an integrated health system designed to provide near-equal access to care, remains uncertain. To evaluate whether overall survival (OS) and cancer-specific survival (CSS) differ between Black veterans and those with other race receiving cancer care through VHA. PubMed was searched from January 2015 through April 2022. Reference lists from identified studies were also reviewed. Studies of US veterans receiving cancer care through the VHA were included if they reported OS or CSS by race and provided hazard ratios (HRs). Dual independent rating of titles and abstracts was conducted for inclusion. A random-effects model was used to pool effect sizes, the Paule-Mandel estimator was used to calculate the heterogeneity variance τ2, and Knapp-Hartung adjustments were used to calculate the confidence interval of the pooled effect. Review and meta-analysis was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guideline. OS and CSS were compared between Black veterans and those who were not Black using pooled HRs. Of 101 studies identified, 39 met inclusion criteria and 29 provided sufficient data for meta-analysis. The reported outcomes represented 603 256 veterans with cancer treated between 1983 and 2017, with a mean 29.0% (range, 8.9%-55.0%) of patients categorized as Black. All studies compared race by Black compared with White, except for 7 of 20 prostate cancer studies, which compared race by Black compared with non-Black. Black veterans were found to have better OS (HR, 0.93; 95% CI, 0.89-0.97) and CSS (HR, 0.94; 95% CI, 0.90-0.98). Survival advantages for Black veterans were observed across several cancer types, including bladder, laryngeal, lung, oropharyngeal, prostate, and plasma cell cancers. Between-study heterogeneity was low to moderate. In this systematic review and meta-analysis of peer-reviewed publications reporting the outcomes of veterans receiving cancer care through VHA, survival outcomes were generally similar or better for Black compared with White or non-Black veterans. These findings suggest that integrated health care systems providing near-equitable access to comprehensive cancer care can substantially reduce or eliminate disparities in cancer outcomes.

Food insecurity affects nearly 1 in 5 US adults and is associated with barriers to preventive health care. Whether food insecurity is associated with lower national cancer screening adherence and whether Supplemental Nutrition Assistance Program (SNAP) participation is associated with attenuation of these disparities remain unclear. To evaluate the association between food insecurity and guideline-concordant colorectal, breast, and cervical cancer screening and to assess whether SNAP participation was associated with attenuation of screening disparities among food-insecure adults. This cross-sectional study used 2022 Behavioral Risk Factor Surveillance System data, a nationally representative survey of noninstitutionalized US adults. The study took place from May 22 to July 21, 2025. Data analyses were conducted from June 3 to July 7, 2025, and included respondents with complete food insecurity, SNAP participation, covariate, and age- and sex-eligible cancer screening data. Self-reported food insecurity during the past 12 months; SNAP participation served as the primary effect modifier. Guideline-concordant colorectal, breast, and cervical cancer screening based on US Preventive Services Task Force recommendations. Of 251 107 adults included in the analytic cohort, 47 453 (18.9%) reported food insecurity; food-insecure respondents were more often younger than 65 years (35 031 [73.8%] vs 118 063 [58.0%]) and female (27 690 [58.4%] vs 106 203 [52.2%]) compared with food-secure respondents. Screening adherence was lower among food-insecure than food-secure adults for colorectal (10 115 [51.6%] vs 59 825 [63.8%]), breast (1459 [61.2%] vs 7457 [72.5%]), and cervical cancer screening (1124 [39.8%] vs 4682 [50.3%]). In adjusted analyses stratified by SNAP participation, food insecurity was associated with lower odds of colorectal (adjusted odds ratio [aOR], 0.78; 95% CI, 0.74-0.83), breast (aOR, 0.69; 95% CI, 0.60-0.79), and cervical cancer screening (aOR, 0.63; 95% CI, 0.49-0.81) among respondents not enrolled in SNAP. Among food-insecure respondents, adjusted probabilities of being up to date with screening were higher among SNAP participants than nonparticipants for colorectal (48.2% [95% CI, 46.1%-50.2%] vs 46.3% [95% CI, 44.1%-48.4%]; difference, 1.9 [95% CI, 0.5-3.6] percentage points), breast (56.1% [95% CI, 54.0%-58.3%] vs 52.7% [95% CI, 49.7%-55.8%]; difference, 3.4 [95% CI, 1.3-5.6] percentage points), and cervical (51.3% [95% CI, 48.9%-53.8%] vs 45.2% [95% CI, 42.5%-47.4%]; difference, 6.1 [95% CI, 3.8-8.4] percentage points) cancer screening. In this cross-sectional study of 251 107 US adults, food insecurity was associated with lower likelihood of guideline-concordant colorectal, breast, and cervical cancer screening. Among food-insecure adults, SNAP participation was associated with higher adjusted screening probabilities than nonparticipation.

The increasing global burden of cancer necessitates innovative therapeutic strategies. Cell therapy represents a major breakthrough in oncology, evolving rapidly from the successful application of chimeric antigen receptor T (CAR-T) cells in hematologic malignancies to a multiplatform landscape characterized by the concurrent development of diverse strategies. Current research focuses on T cell receptor-engineered T (TCR-T) cells, tumor-infiltrating lymphocytes (TILs), gamma delta (γδ) T cells, CAR-natural killer (CAR-NK) cells, CAR-macrophages (CAR-Ms), and various strategies based on dendritic cells (DCs), B cells, and stem cells. The translational paradigm is expanding from the relatively mature field of hematologic malignancies to the more prevalent and mechanistically complex domain of solid tumors. In recent years, this field has exhibited a clear trend toward expansion from autologous therapies to allogeneic "off-the-shelf" platforms. Approaches such as CAR-NK and CAR-natural killer T (CAR-NKT) cell therapies exhibit significant clinical potential because of their low immunogenicity and reduced risk of graft-versus-host disease (GvHD). Concurrently, in vivo engineering technologies that directly deliver CAR genes in situ are emerging as promising approaches to lower costs and simplify manufacturing by bypassing complex ex vivo procedures. This review systematically outlines recent advances in these strategies, focusing on their mechanisms of action, target antigens, and clinical translation. Despite progress, formidable challenges remain, including tumor heterogeneity, the immunosuppressive tumor microenvironment (TME), and therapy-related toxicity. To address these challenges, future research will focus on novel target discovery, enhanced toxicity management, and scalable manufacturing processes. The integration of multidisciplinary technologies, such as multiomics analysis, artificial intelligence, and synthetic biology, will advance cell therapies toward safer, more effective, and widely accessible applications.

Objectives. To examine associations of ultraprocessed foods (UPFs, defined by Nova) with cardiometabolic risk factors, prevalent conditions, and mortality, before and after comprehensively adjusting for nutritional quality. Methods. We analyzed data from 47 999 adults in the National Health and Nutrition Examination Survey (NHANES) 1999-2018. Survey-weighted, multivariable adjusted linear or logistic regression evaluated cross-sectional associations with risk factors and disease prevalence, and Cox models evaluated prospective associations with all-cause mortality. Models were compared before and after adjustment for each individual's Food Compass Score (i.FCS) to test independence from nutritional quality. Results. Every 10% of energy supplied from UPFs was associated with higher body mass index, HbA1c, diastolic blood pressure, total-to-high-density lipoprotein cholesterol (HDL-C); lower HDL-C and low-density lipoprotein cholesterol (LDL-C); greater prevalence of metabolic syndrome (odds ratio [OR] = 1.07; 95% confidence interval [CI] = 1.05, 1.09), diabetes (OR = 1.03; 95% CI = 1.00, 1.07), and cancer (OR = 1.05; 95% CI = 1.02, 1.08); and higher risk of all-cause mortality (hazard ratio = 1.04; 95% CI = 1.02, 1.07). When we adjusted for i.FCS, associations were only partly attenuated, remaining significant. By comparison, adjustment for saturated fat, added sugar, or sodium had little effect. Findings were consistent in population subgroups, except for stronger associations among lower-income adults. Conclusions. UPF consumption is associated with adverse risk factors, disease conditions, and all-cause mortality, only partly explained by nutritional quality. (Am J Public Health. 2026;116(7):1015-1024. https://doi.org/10.2105/AJPH.2026.308499).

Human papillomavirus (HPV) is the most common sexually transmitted infection worldwide and is responsible for anogenital warts and several cancers. Although the effectiveness of female HPV vaccination has been well established since its introduction in 2006, no systematic review has specifically evaluated the impact of male vaccination programs introduced in 2011. We synthesised real-world evidence on gender-neutral vaccination (GNV), identifying 17 studies reporting rates of HPV infection, anogenital warts, and HPV-related cancers during the pre- and post-GNV periods among males in high-income countries. Overall, we found limited but consistent evidence of benefit in males, although some of the observed reductions may also reflect herd immunity effects from earlier female-only HPV vaccination programs. Only two studies evaluated temporal trends in HPV infection, both demonstrating reductions in vaccine-targeted genotypes, although important methodological limitations restrict generalisability. Eight studies reported consistent declines in anogenital warts among adolescents and young adults, supporting a beneficial population-level impact of GNV. Analyses of HPV-related cancers showed an overall increasing incidence, likely reflecting long latency periods. However, some studies reported encouraging declines in cancer incidence among younger age groups. Oropharyngeal incidence decreased by 11%-19% among U.S. males aged 20-44 years, and penile cancer incidence decreased by 6%-38% among those aged 15-34 years. In Texas, penile and anal cancer incidence decreased by 35% and 16%, respectively, among males 20 years or older, while in Austria, anal cancer incidence decreased by 38% among men aged 30-39 years. Given the relatively recent introduction of male vaccination programs, longer follow-up is required to fully assess their impact. Studies in low- and middle-income countries are urgently needed.

Data unveiled at this year's ASCO Annual meeting show that GRAIL's Galleri multi-cancer early detection (MCED) test failed to reduce stage III and IV cancers in the landmark NHS-Galleri trial, although there was an improvement in stage IV alone. Experts say the findings fall short of demonstrating a large enough clinical benefit for MCED testing that would warrant Galleri's implementation.

Cellular metabolism, redox balance, and viability are tightly constrained by oxygen availability. Reduced oxygen tension activates a conserved hypoxic signaling network centered on hypoxia-inducible factors (HIFs), which coordinate transcriptional and post-transcriptional programs that promote metabolic adaptation, angiogenesis, erythropoiesis, and redox balance. While HIF signaling is essential for development and tissue homeostasis, its dysregulation contributes to disease-associated dysfunction in cancer, ischemic disease, and chronic inflammatory disorders. Accordingly, therapeutic strategies aimed at modulating this pathway have gained considerable interest. This review critically examines current strategies aimed at suppressing HIF signaling, with a particular emphasis on approaches that reduce HIF-α protein abundance, inhibit transcriptional activity, or interfere with HIF complex formation. We discuss RNA-based therapies, small-molecule modulators of HIF stability, and noncanonical HIF inhibitors - including topoisomerase inhibitors, proteasome inhibitors, and metabolic modulators - that exert their effects independently of classical oxygen-dependent degradation. Emerging evidence indicates that many chemically diverse HIF inhibitors converge on translation-centered mechanisms, including inhibition of mTOR signaling, activation of eIF2α-dependent stress responses, and disruption of hypoxia-specific translational control. These pathways represent a dominant and previously underappreciated layer of HIF regulation. By contrasting stability-, translation-, and transcription-centered strategies, we highlight key therapeutic constraints imposed by oxygen availability, redox balance, and HIF isoform specificity. The clinical success of HIF-2α-specific allosteric inhibitors underscores the importance of exploiting unique structural vulnerabilities, whereas the lack of analogous pockets in HIF-1α necessitates alternative approaches. We propose that durable suppression of hypoxia-driven pathology will likely require integration of isoform-specific targeting with translational control mechanisms that decouple HIF signaling from generalized cytotoxic stress.

Subway particulate matter (PM) pollution represents an increasingly important environmental health issue, yet its spatial variability and exposure implications across subway microenvironments remain insufficiently synthesized. This study reviewed 196 studies from 24 countries spanning a 15-year period, to characterize mass concentrations and chemical composition of PM10, PM2.5, and PM1 across key subway spaces, including station offices, concourses, platforms, tunnels, carriages, and commuting exposure, and to quantify associated health risks. Subway PM exhibited pronounced spatial heterogeneity: concentrations were highest in tunnels, followed by platforms and commuting exposure, whereas concourses and carriages showed comparatively lower but still elevated levels. Limited evidence for office environments nevertheless indicated severe pollution, with peak concentrations reaching 405.1 µg/m3 for PM10 and 212.1 µg/m3 for PM2.5, suggesting an overlooked occupational exposure scenario. Strong enrichment of metal components was observed in subway aerosols. Fe showed the highest indoor-to-outdoor ratios, with mean values up to 66.5 for PM2.5 and 41.5 for PM10. Multi-study synthesis identified key drivers of PM distribution, including station depth, platform door configuration, operation frequency, ventilation, season, and outdoor pollution, reflecting combined internal generation and external infiltration. Health-risk assessment revealed that carriages constituted the highest-risk microenvironment. Mean cancer risks reached 8.4 × 10-4 for PM2.5 and 4.5 × 10-4 for PM10, while non-cancer risks were 28.0 and 2.4, respectively. Subway employees experienced greater risks than passengers. Element-specific analysis showed carcinogenic risk was primarily driven by Cr and Co, whereas non-carcinogenic risk was mainly associated with Co, Ni, and Mn, providing a basis for microenvironment-targeted air quality control in subway systems.

Multi-modal data-based algorithms have gained attention in their capabilities in prediction tasks in cancer-related research. This paper introduces SPACT, a multi-modal capable of predicting cancer survival probability based on a deep-learning application on the histopathological patch features, whole-slide histopathology (WSI) in particular. What SPACT improves upon compared to the existing survival-prediction models is aiming to improve the robustness/resilience and the prediction accuracy, both overall and in subcategories. It does that by instead of relying only on the TCGA-based datasets, it benefits from the use of an external dataset collected from the Başkent Hospital. By doing cross-comparison on the different encoders' accuracy on both datasets, an optimal encoder that performs well in both the conventional TCGA and newly gathered Başkent Hospital dataset is chosen. Using a combination of multiple encoders on different models, we find that encoders that perform well on both datasets outperform the encoders that work well on a single one. Additionally, in-depth analyses on the ablation studies, risk stratification, attention maps, and on how integrated gradients may have an effect on the performance and reasoning of SPACT. SPACT matches or outperforms all the other state-of-the-art multi-modal prediction algorithms in 5 of the 7 different cancer types, particularly in the case of ovarian cancer, by a wide margin, with a c-index score of 0.77. It has the highest robustness, having the best overall performance on all 3 encoders the models have been tested with. These findings solidify SPACT's position as a contemporary and improved multi-modal based deep learning model targeting individual prediction accuracy, overall prediction accuracy, and robustness under diverse conditions. The code for SPACT is available at https://github.com/ezgiogulmus/SPACT.

Gastrointestinal (GI) tract cancers are the second leading cause of cancer-related mortality, often due to late detection. There is a critical need for non-invasive, highly sensitive biomarkers for early-stage cancer and precancerous lesion detection to enable timely intervention. This study aimed to develop a blood-based method specifically optimized for early GI cancer detection and population-level screening. Using large-scale public tissue methylation data and the Twist probe cfDNA profiles, we developed SPOGIT (Screening for the Presence of Gastrointestinal Tumors), a multi-algorithm model (Logistic Regression/Transformer/MLP/Random Forest/SGD/SVC) for early GI cancer detection. The model was rigorously validated through an internal (n = 83) and multicenter external validation (386 cancers/113 controls/580 precancers), with an interception model assessing its clinical potential. SPOGIT demonstrated high accuracy in detecting GI cancers, with a sensitivity of 88.1 % and a specificity of 91.2 %. Notably, it effectively identified early-stage (0-II) cancers with 83.1 % sensitivity. The model also showed significant potential for intercepting premalignant progression, detecting advanced adenomas (AA) and gastric precancerous lesions with sensitivities of 56.5 % and up to 62.4 %, respectively. In the external independent validation cohort, a complementary model CSO (Cancer Signal Origin) demonstrated an accuracy of 83 % for colorectal cancer and 71 % for gastric cancer. Most importantly, simulation analyses projected that SPOGIT implementation could significantly reduce late-stage diagnoses and increase 5-year survival rate by 27.02 % through early interception. This study introduced a novel, dual-model blood architecture (SPOGIT/CSO) that enables highly accurate, early detection of GI cancers and their precursors. By facilitating timely clinical intervention, SPOGIT/CSO represented a paradigm-shifting strategy with the potential to significantly improve patient survival outcomes and transform GI cancer management.

Urinary system tumors, including bladder, kidney, and prostate cancers, impose a substantial health burden worldwide and exhibit significant molecular and genetic heterogeneity. of Review: This review delves into the intricate landscape of targeted therapy for urinary system tumors and its pivotal role in advancing precision medicine. Key Scientific Concepts of Review: Urinary system tumors include kidney cancer, bladder cancer, and prostate cancer. They exhibit remarkable heterogeneity at the molecular level, which challenges the effectiveness of conventional treatments. Compared with conventional therapy, precision medicine is a tailored approach that customizes treatments for individual patients on the basis of their unique genetic, molecular, and clinical profiles. In general, precision medicine discovers targets through multiomics data, and targeted therapy develops drugs on the basis of these targets. Targeted therapy focuses on the discernment of specific molecular or genetic aberrations that are distinct to cancer cells, thereby demarcating them from their healthy counterparts. Several molecular targets driving these malignancies are summarized among the current state of approved targeted therapies. Limitations include the emergence of resistance mechanisms and the potential for adverse effects. In the pursuit of precision medicine, this review scrutinizes innovative approaches, including biomarker utilization, genomics, and personalized treatments, which demonstrate potential in customizing therapies to individual patients. In conclusion, this review illuminates the pivotal role of targeted therapy in the context of heterogeneous urinary system tumors and underscores its potential to drive the precision medicine paradigm forward.

Rural-urban disparity of cancer is a major public health problem, with an unclear gap in noncancer death. It is important to evaluate rural-urban disparities in cardiovascular diseases (CVDs) and other competing death among cancer patients. Observing urban-rural disparities and trends in noncancer deaths in the U.S. cancer population. To address rural-urban disparities, we used proportions of deaths, age-adjusted mortality rates (AAMR), cumulative mortality rates, subdistribution hazard ratio (sHR), standardized mortality ratios (SMRs), absolute excess risks (AERs) and mediation analysis. Between 1990 and 2017, there were 2,022,482 patients of 24 cancer sites, with a median follow-up of 11·8 years. Rural proportions of noncancer and CVD deaths in cancer patients were higher than urban ones. Rural AAMR of noncancer and CVD deaths surpassed urban one in cancer patients, who had higher cumulative mortality rates than urban counterparts in noncancer (sHR:1·21, 95 % confidence interval [CI]:1·19-1·22), CVDs (sHR:1·25, 95 % CI: 1·23-1·28), diabetes mellitus (sHR:1·24, 95 % CI:1·15-1·34), Alzheimer's disease (sHR:1·30, 95 % CI:1·21-1·39), pneumonia and influenza (sHR:1·30, 95 % CI:1·21-1·39) and chronic obstructive pulmonary disease and allied cond (sHR:1·32, 95 % CI:1·26-1·39). Compared with the general population, both rural (SMR:4·58, AER:218·03) and urban (SMR: 3·74, AER: 166·61) cancer patients had higher risks of noncancer death. Mediation analyses identified median household income and SEER stages as the mediators. Rural cancer patients had higher risks of noncancer death than urban counterparts, especially CVD-related deaths. Future targeted policy and public health interventions are needed to diminish the rural-urban gap in such death disparities.

Large language models (LLMs) show strong performance on clinical benchmarks, yet their reasoning reliability in real-world oncology care remains unclear. We evaluated LLM reasoning on authentic oncology notes using a novel hierarchical error taxonomy across two retrospective cohorts spanning breast, pancreatic, and prostate cancer. GPT-4 produced reasoning errors in 23.1% of note interpretations, the majority reflecting cognitive bias patterns. Errors were more frequent in recommendation tasks and were strongly associated with guideline-discordant recommendations and lower clinician-rated clinical impact scores. Confirmation bias, anchoring bias, and omission errors were most strongly linked to potentially harmful outputs. Compared to GPT-4, GPT-5.1 demonstrated reduced error rates and improved clinical performance but retained structured reasoning failure patterns. Automated LLM-based evaluators detected error presence but failed to reliably classify subtypes, and a preliminary self-mitigation strategy yielded only modest improvement. Endpoint accuracy alone may mask clinically meaningful reasoning failures. Therefore, evaluating and monitoring reasoning fidelity should be a prerequisite for safe deployment of LLMs in oncology decision support.

Precision oncology is revolutionizing cancer care, but isn't reaching enough patients. Molecular tumor boards (MTBs) translate complex genomic data to more effectively inform personalized care, and a new meta-analysis shows they boost clinical outcomes. But how can we feasibly and equitably expand the use of MTBs?

As cancer survival rates improve, the long-term burden of treatment has become increasingly evident. Cancer survivors face a markedly higher risk of cardiovascular-related complications and premature, non-cancer-related mortality. In particular, cardiovascular disease (CVD) is disproportionately prevalent, with survivors approximately 40% more likely to develop and die from CVD compared to the general population. Although this increased morbidity reflects both acute cardiotoxic events and the later development and progression of more chronic CVDs, traditionally viewed as direct consequences of cancer therapies, the underlying mechanisms, especially those that are feasible to modify, are poorly understood. Emerging evidence positions the gut-heart axis as a central regulator of cardiovascular risk in cardio-oncology. This avenue is especially compelling to explore as the gut microbiome is well documented to be altered by cancer therapies, including chemotherapy, immune checkpoint inhibitors, targeted therapies, and radiation, with profound and persistent changes in diversity, composition and function widely reported across clinical cohorts. Comparable microbial changes have been observed in non-cancer cohorts with cardiovascular disease. For example, specific microbial metabolites have been reported to exert cardiovascular protective benefits in hypertension. Thus, there is a compelling opportunity to explore the gut microbiome to advance our understanding and ability to prevent cardiovascular disease in cancer survivors. This review synthesises current evidence linking the gut microbiome to cancer therapy-related cardiac dysfunction (CTRCD), evaluates microbial metabolites as predictive biomarkers of cardiotoxicity, and discusses microbiome-targeted modulation as an emerging strategy for improving cardiovascular outcomes in cancer survivors.

Cancer-related sarcopenia is associated with poor clinical outcomes but remains difficult to define and quantify in routine oncology practice. Current assessments rely on imaging and functional scales that are time-consuming and provide limited biological insight. We aimed to identify a plasma proteomic signature of cancer-related sarcopenia and to uncover circulating mediators involved in its pathophysiology. Patients were included from two cohorts of the MATCH-R study (NCT02517892): a discovery cohort of advanced cancer patients treated with immunotherapy and an independent validation cohort of metastatic castration-resistant prostate cancer (mCRPC) patients treated with androgen-receptor pathway inhibitors. External validation was performed in the TRACERx cohort of non-small cell lung cancer. Skeletal muscle index at third lumbar vertebra (L3) was quantified using imaging, and ECOG performance status served as a functional proxy. Plasma proteomics was performed using the Olink Explore platform. An extreme gradient boosting (XGBoost) model was trained on a high-contrast subset using a neuromuscular-focused protein panel and validated across cohorts. Functional effects of candidate mediators were assessed in differentiating human myoblasts. The model generated a continuous sarcopenia probability (SP) score that correlated with muscle mass and functional status and consistently stratified overall survival across cohorts. A reduced four-protein model retained comparable performance, supporting translational applicability. Proteins associated with SP included insulin-like growth factor binding protein 1 and 2 (IGFBP1, IGFBP2), and interleukin-6 (IL6). IGFBP1 and IGFBP2 impaired myoblast differentiation, while IL6 induced IGFBP1 expression in liver cells. Plasma proteomics enables scalable and biologically informed assessment of cancer-related sarcopenia, identifies tumor-host mediators of muscle dysfunction, and supports objective patient stratification for therapeutic intervention.

This study maps the global clinical trial landscape of cytokine-induced killer (CIK) cell therapy across malignancies and selected immune-related disorders, based on 199 interventional trials retrieved from the Informa database up to February 2026. Analysis reveals a rapidly expanding field dominated by early-phase studies, with China leading clinical translation and academic institutions as primary sponsors. Oncology trials are most frequent, particularly in lung cancer, leukemia, and hepatocellular carcinoma, while combinatorial strategies such as dendritic cell-CIK therapy or checkpoint inhibitor combinations are increasingly explored. Adjuvant and maintenance settings predominate over neoadjuvant applications, and emerging studies indicate potential extension into autoimmune and inflammatory diseases. These findings underscore the translational potential of CIK therapy, highlight evolving combination strategies, and reveal opportunities to optimize patient stratification, standardize cell preparation, and incorporate biomarker-guided approaches. Collectively, this landscape analysis highlights key trends shaping the future clinical development of CIK-based immunotherapy and informs strategic design considerations for advancing its integration into modern oncology and immunomodulatory treatments.

Although A2AR is a key immunoregulatory receptor that suppresses CD8+ T cell activation in response to elevated extracellular adenosine in inflamed or hypoxic microenvironments, its role in CD8+ T cell differentiation and cell-fate decisions during chronic viral infection and cancer remains poorly understood. Using A2AR-eGFP reporter mice, we show that A2AR expression is rapidly induced by TCR stimulation and persists under chronic antigen exposure and hypoxia, with sustained expression strongly associated with terminal exhaustion via the canonical Gαs-cAMP-PKA pathway. Paradoxically, A2AR loss does not alleviate exhaustion but instead accelerates differentiation toward the terminally exhausted state. Single-cell multiomics profiling revealed that A2AR deficiency activates CD122 (IL-2Rβ)-dependent signaling, driving T cell exhaustion. Genetic deletion of CD122 in A2AR-deficient CD8+ T cells reduced terminal exhaustion, identifying CD122 signaling as a key mediator of A2AR loss-driven exhaustion. Intriguingly, both sustained A2AR expression and A2AR loss converge to promote T cell exhaustion differentiation through distinct mechanisms. These findings uncover a paradoxical role of A2AR in shaping CD8+ T cell fate choices during chronic infection and cancer.

Cyclins are fundamental regulators of the eukaryotic cell cycle, orchestrating cell cycle transitions through their interactions with cyclin-dependent kinases (CDKs). Aberrant expression or activation of cyclins disrupts normal cell cycle control, leading to uncontrolled proliferation, genomic instability, and tumorigenesis. Overexpression of specific cyclins has been documented in multiple malignancies, notably breast cancer, underscoring their oncogenic potential. Despite extensive research, the context-dependent roles of cyclins, their isoform-specific functions, and mechanisms underlying resistance to cyclin-targeted therapies remain incompletely understood. This review aims to provide an updated and comprehensive overview of the molecular mechanisms by which cyclins contribute to cancer initiation and progression. It also explores recent advances in therapeutic interventions targeting cyclins and CDK signaling pathways, highlighting both the promise and limitations of current cyclin-directed strategies. We systematically analyzed relevant literature retrieved from PubMed and Google Scholar using keywords such as "cyclins," "cyclin-dependent kinases (CDKs)," "cell cycle," and "CDK inhibitors." The review discusses the multifaceted roles of cyclins in promoting oncogenic transformation and therapeutic resistance, emphasizing the importance of cellular context in determining their biological effects. Furthermore, it evaluates novel approaches to overcome resistance to CDK4/6 inhibitors and the potential of cyclin-based therapeutic targeting to improve cancer management and patient outcomes. A deeper understanding of cyclin biology may facilitate the development of precision strategies against cancer metastasis and progression.

The oral microbiome is comparable to the gut microbiome in ecological complexity and is now recognized as a contributor to anticancer immune responses. Although the relationship between the gut microbiome and anticancer immunity is well established, the connection between the oral microbiome and anticancer immunity has received increasing attention, with accumulating evidence pointing to the direct effects of the oral microbiome on immune cell populations. The relationship between cancer and the oral microbiome is bidirectional: each influences the behavior of the other. The tumor microenvironment (TME) and oncological therapies such as chemotherapy and radiation can cause oral microbiome dysbiosis. Once dysbiosis is established, it creates conditions that favor tumor initiation and recurrence through chronic inflammation and impaired immune surveillance. Furthermore, the oral microbiome indirectly affects distant cancers and contributes to systemic inflammation and microbial dissemination through gastrointestinal, respiratory, hematogenous, neurological, and lymphatic pathways. Prebiotics, probiotics, postbiotics, and microbiota transplantation represent promising therapies targeting this microbial community to enhance the efficacy of cancer immunotherapy.

A substantial fraction of human long noncoding RNAs (lncRNAs) are lineage specific and increasingly implicated in cancer, yet the evolutionary origins of their oncogenic relevance remain unclear. It is unknown whether such roles arise through adaptive refinement or as incidental by-products of transcriptomic complexity under weak selection. Here, we show that newly evolved cancer-associated lncRNAs, predominantly of primate origin, arise through a stepwise process initiated by increased transcriptional activity. Once expression surpasses a threshold, these lncRNAs undergo sequence expansion, with 73.3% incorporating external or local genomic fragments, facilitating integration into deeply conserved cancer-related pathways. We validate this model through MIR497HG, which originated near a microRNA in the human-macaque ancestor, and subsequently expanded and integrated into the ancient AMPK (adenosine monophosphate-activated protein kinase)/ferroptosis network in humans, influencing oncogenesis. Collectively, our findings support a model in which weak selection permits neutral transcriptional innovations to persist and gradually infiltrate ancient regulatory architectures, consistent with constructive neutral evolution.

Metabolic crosstalk between cancer cells and immune cells is now recognized as a major determinant of immune escape and resistance to anticancer treatments. Cancer cells profoundly reshape the metabolic landscape of the tumor microenvironment, driving nutrient competition, hypoxia, and the accumulation of immunosuppressive oncometabolites that collectively blunt antitumor immunity. Effector T cells, NK cells, and dendritic cells are exposed to nutrient deprivation and suppressive metabolites, including lactate, adenosine, and kynurenine, resulting in impaired T cell proliferation and cytotoxic function and expansion of metabolically adapted regulatory T cells and myeloid-derived suppressor cells. Cancer-associated fibroblasts further reinforce this metabolic reprogramming through extracellular matrix remodeling, secretion of immunosuppressive metabolites, and nutrient recycling that supports tumor growth. Abnormal tumor vasculature sustains metabolic stress by causing uneven perfusion, hypoxia, and acidosis, thereby limiting immune cell infiltration, and promoting immune exhaustion. In addition, diet- and microbiome-driven metabolic cues dynamically shape cancer-immunity interactions and therapeutic responses. Targeting key metabolic checkpoints, including glycolysis, adenosine signaling, tryptophan metabolism, fatty acid oxidation, and lactate production, has emerged as a promising strategy to restore antitumor immunity. Nevertheless, metabolic heterogeneity, context-dependent immune responses, and safety concerns pose persistent challenges to its successful implementation. Recent advances in biomarker development, patient stratification, and rational combination strategies underpin the clinical translation of metabolic-immune vulnerabilities in cancer therapy. Integrating metabolic interventions with immune checkpoint blockade or adoptive cell therapies has demonstrated synergistic effects in preclinical and early clinical studies, enhancing T cell persistence and cytotoxic function within metabolically hostile tumor microenvironments. This review addresses these issues and delineates the mechanistic basis of the dynamic interplay between cancer metabolism and immune regulation. It discusses how anti-cancer therapies affect metabolic and immune pathways and highlights next-generation, metabolically targeted therapies that leverage newly uncovered, tumor-specific rewiring of glycolysis, mitochondrial function, and nutrient uptake. Special emphasis is given to the development of first-in-class inhibitors targeting glutaminase, lipid biosynthesis, one-carbon pathways, and redox homeostasis, which, when paired with immunotherapy or conventional treatments, offer unprecedented opportunities to overcome metabolic barriers, abrogate resistance, and achieve durable immune control of cancer.

Tobacco use is linked to multiple cancer types and a quarter of cancer deaths. Tobacco smoke contains numerous mutagenic chemicals such as polycyclic aromatic hydrocarbons (PAH), however, the mutagenicity of many chemicals in tobacco products remains underexplored. The tobacco-specific nitrosamines 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN), both International Agency for Research on Cancer (IARC) Group 1 carcinogens, form pre-mutagenic pyridyloxobutyl (POB) DNA adducts, yet little is known about the associated genomic mutation landscape(s) in humans. We applied bulk and error-corrected next-generation sequencing to a battery of experimental systems comprising mammalian cell lines and animals exposed to NNK or NNN, and the extracted mutational signatures were compared to POB DNA adducts formed in the same models. A genome-scale POB mutational signature was extracted and characterized in detail and subsequently used for optimized per-sample signature attribution in ~ 6,670 cancer genomes from the International Cancer Genome Consortium Pan-Cancer Analysis of Whole Genomes (ICGC PCAWG) and The Cancer Genome Atlas (TCGA) collections. POB signatures generated in experimental exposure models share profiles dominated by T > N and C > T alterations, with transcription strand asymmetry indicative of damage on thymidines and cytidines. The T > N-dominated mutation profiles can be explained by O2-POB-dT adducts, detected in the exposed cells. Stringent screening of 6,668 cancer genomes revealed the presence of a genome-scale POB signature in ~ 350 tumors from sites associated with tobacco smoking, that were distinct from cancer sites typically linked to known smoking-related single base substitution (SBS) signatures SBS4 and SBS92 linked to PAHs. These included hematological malignancies and cancers of the kidney, prostate, pancreas and breast, among other sites. In contrast and as expected, tobacco-smoking signatures SBS4 and SBS92 were jointly or individually predominant in cancers of the lung, liver, head-and-neck or bladder. Our study provides first-of-its-kind evidence for the presence of a tobacco-specific nitrosamine mutational signature in human tumors. It suggests that the POB pathway may contribute selectively to the mutational landscapes of certain cancers distinct from those affected by PAH compounds, indicating the potential roles of specific tobacco smoke chemicals in tobacco-linked cancer types lacking SBS4 or SBS92.

Oncolytic viruses (OVs) represent a versatile platform for cancer immunotherapy, capable of selectively infecting and lysing tumor cells while triggering systemic antitumor immunity. However, their therapeutic efficacy remains limited by antiviral immunity, restricted intratumoral spread, and an immunosuppressive tumor microenvironment. This review highlights emerging strategies to potentiate OV efficacy through rational combination with complementary immunotherapies, including immune checkpoint inhibitors, adoptive cell therapies, cancer vaccines, and small-molecule immunomodulators. These synergistic interventions can remodel the tumor microenvironment, enhance immune cell infiltration and activation, reverse immunosuppressive feedback, promote immunogenic cell death, normalize the tumor vasculature, and modulate the gut microbiota, collectively amplifying OV replication and oncolytic potency. The integration of artificial intelligence and multiomics profiling further enables precise patient stratification and optimization of combination regimens. In parallel, advanced engineering strategies, such as arming OVs with immunomodulatory transgenes and mitigating host antiviral responses, further reinforce these effects. Together, these approaches illustrate how multimodal immunotherapy can overcome the intrinsic limitations of OVs, enabling durable antitumor immunity. This review underscores the central role of combination strategies in OV-based immuno-oncology and outlines future directions to accelerate their clinical translation.

Dysregulation of cell death is a hallmark of cancer, and overcoming tumor resistance to apoptosis-inducing therapies remains challenging. Inducing immunogenic cell death (ICD), such as pyroptosis, is promising but currently lacks tools for in vivo monitoring and control. Here, we developed a caspase-3-activated, genetically encoded protein expression system-a programmable synthetic biology platform activated by the key apoptotic protease caspase-3-for apoptosis detection. Configured to express fluorescent or luciferase proteins, it enabled high-resolution live-cell apoptosis imaging and in vivo tumor bioluminescence imaging, and it served as a screening tool for proapoptotic microRNAs. Most importantly, engineering it to express a pyroptosis effector triggered apoptosis-coupled pyroptosis, achieving potent cancer cell killing and enhanced in vivo antitumor efficacy. This versatile platform advances apoptosis detection and its conversion to potent ICD, offering an adaptable approach for cancer theranostics.

Itaconate (ITA) is an endogenous metabolite produced from cis-aconitate by the immune-responsive gene 1-encoded enzyme aconitate decarboxylase 1 and has emerged as an important immune-metabolic regulator in cancer. In the tumor immune microenvironment, endogenous ITA is increasingly associated with an immunosuppressive microenvironment, tumor cell immune evasion, and therapeutic resistance, supporting a predominantly pro-tumor role in vivo. Nevertheless, the field remains clouded by several unresolved issues, including the heavy reliance on a limited number of tumor models, the persistent conflation of endogenous ITA with membrane-permeable derivatives, the incomplete definition of the mechanisms underlying its intercellular effects, and the uncertain cross-species relevance of current findings. Here, we highlight recent evidence supporting the physiological functions of endogenous ITA in cancer, delineate major boundaries and ambiguities in current interpretation, and outline key questions that should guide future mechanistic and translational studies.

Interleukin-33 (IL-33) is classically viewed as an alarmin, but in cancer, it may do more than merely initiate immunity. I propose that IL-33 scripts the trajectory of immunity through interconnected regulatory layers that influence how host responses are organized and ultimately resolved into either productive antitumor immunity or suppressive, repair-like states.

Drug resistance, defined as the adaptive tolerance of human diseases and agricultural pests to therapeutic or chemical agents, threatens global health and food security. Alternative splicing (AS), a central posttranscriptional regulatory mechanism, is increasingly recognized as a conserved driver of resistance evolution. However, although AS-mediated resistance has been widely studied in cancer, its role in pesticide and fungicide resistance remains insufficiently explored, limiting cross-disciplinary reuse of mechanistic insights and intervention strategies. This review critically compares the spliceosome-mediated and AS-mediated mechanisms that underlie resistance to anticancer drugs, pesticides and fungicides within a unified framework, with the goal of clarifying how distinct selective pressures shape convergent adaptive resistance across biological kingdoms. We summarize how aberrant AS promotes resistance through target truncation, altered drug metabolism, apoptosis evasion, and rewiring of survival signaling. We further propose a customized Design-Build-Test-Learn (DBTL) roadmap for AS-targeted intervention, integrating long-read sequencing, AI-assisted prediction, splice-modulating therapeutics such as ASOs and CRISPR-based approaches, nanodelivery optimization, and dynamic resistance monitoring. By linking mechanistic comparison with translational implementation, this review provides an interdisciplinary framework for understanding, predicting, and reversing AS-mediated resistance in both clinical and agricultural settings.

Biguanides, including metformin, the world's most prescribed oral hypoglycemic, extend health span and lifespan in vertebrates and invertebrates. Given the widespread use and apparent safety of metformin, it is assumed that its effects are not associated with toxicity, except when in marked excess. Here, we determine that accumulation of damaging reducing equivalents is an unanticipated toxicity associated with biguanides, defense against which requires post-transcriptional protection of de novo lipogenesis. We demonstrate that biguanide treatment during impaired lipogenesis drives NADPH toxicity, leading to catastrophic elevation of NADH/GSH reducing equivalents and accelerated death across metazoans. Multiple NADPH-generating interventions require de novo lipogenesis to prevent markedly shortened survival, indicating that this defense mechanism is broadly leveraged. We propose that fatty acid biosynthesis is a tunable rheostat that can minimize biguanide-induced reductive stress while maximizing its pro-longevity outcomes and can serve as an exploitable vulnerability in reductive stress-sensitive cancers.

Sex differences in antitumor immunity contribute to disparities in cancer outcomes, yet the role of CD4+ T helper cells in this context remains poorly understood. In this issue, Hunt and colleagues show that sex hormones restrain CD4+ T-cell help in males, weakening antitumor immune responses and promoting tumor progression. This work identifies CD4+ T-cell help as a previously underappreciated driver of sex-biased cancer immunity, with potential implications for sex-informed immunotherapy strategies. See related article by Hunt et al., p. 1068.

CD8+ T-cell exhaustion limits the immune response to tumors because of ineffective T-cell effector functions. Thus, therapies that inhibit T-cell exhaustion are critical for optimizing cancer treatment. Recent studies have implicated epigenetic proteins in T-cell exhaustion. In this study, we identified activating transcription factor 7-interacting protein (ATF7ip) as an epigenetic protein critical for inducing T-cell exhaustion. Loss of Atf7ip in CD8+ T cells resulted in decreased terminal exhaustion and increased numbers of progenitor-exhausted cells in both chronic viral infections and cancer. Given the decreased T-cell terminal exhaustion observed with Atf7ip deficiency in CD8+ T cells, this may be one mechanism that leads to decreased tumor burden. Mechanistically, ATF7ip functions to stimulate the deposition of repressive H3K9me3 at critical immune-effector gene loci, such as Il7r and Il2, leading to enhanced exhaustion. Our data suggest that ATF7ip may be a rational target for deletion in adoptive T-cell therapies to reduce CD8+ T-cell exhaustion.

Somatic variant discovery in tumors is crucial for clinical analysis, yet most existing methods are designed for short-read sequencing, with few developed specifically for long reads. Here we present Clair-Somatic (ClairS), a deep-learning-based somatic small-variant caller designed for long-read tumor-normal pairs. Trained on synthetic somatic variants with diverse coverages and variant allele fractions, ClairS accurately detects a wide range of somatic variants. Using the Nanopore Q20+ HCC1395-HCC1395BL dataset at 50/25× tumor/normal coverage, ClairS achieved F1 scores of 89.83% for single-nucleotide variations and 73.38% for indels; augmenting training with real cancer cell lines improved performance to 96.19% and 79.67%, respectively. Our findings indicate that improved read phasing enabled by long-read sequencing is key to accurate single-nucleotide variation detection, especially at low variant allele fractions. Through experiments across varied coverage, purity, contamination levels, multiple platforms and real cancer cell lines, we demonstrate that ClairS is a robust and reliable caller. ClairS is open source and available at https://github.com/HKU-BAL/ClairS .

Synthetic mRNA therapeutics offer a versatile platform for treating diverse conditions, including cancer and infectious diseases. For delivery into cells, these mRNAs are encapsulated in lipid nanoparticles and commonly incorporate modified ribonucleotides to improve stability, enhance translation and mitigate immune recognition1. N1-Methylpseudouridine (m1Ψ) has become the industry standard for synthetic mRNAs owing to its effectiveness in promoting translation and reducing immunogenicity2. However, recent studies have shown that m1Ψ can compromise translational fidelity, leading to errors such as premature termination and ribosomal frameshifting3-5. Here we reveal N4-acetylcytidine (ac4C) as a functionally distinct alternative to m1Ψ. Across cultured cell lines, primary human monocyte-derived dendritic cells and mouse liver, ac4C suppressed inflammatory responses as effectively as m1Ψ while driving higher protein yields. Single-molecule imaging of translation revealed broadly similar ribosome densities per mRNA for ac4C-modified and m1Ψ-modified transcripts. However, translation elongation with m1Ψ-modified mRNA was nearly twofold slower than with ac4C, which resulted in reduced protein output and increased ribosome collisions that further limited protein production through the engagement of quality-control pathways and +1 frameshifting. These findings underscore the importance of context in designing therapeutic mRNAs and position the translation elongation rate as a key determinant of the efficacy of modified ribonucleotides.

Oncogenic mutations can have fundamentally different consequences depending on the developmental state of the tissue in which they arise. We propose a unifying model in which early-life oncogenic events promote retention, stabilization of transient embryo-fetal transcriptional and epigenetic programs, whereas later-life mutations more often require rewiring, reactivating suppressed oncofetal states through stepwise transcriptional and chromatin remodeling. This temporal dimension helps explain why childhood and adult cancers can follow distinct lineage trajectories, behaviors, and therapeutic responses, even when initiated by the same genetic lesion. Incorporating developmental timing into cancer modeling and risk stratification may improve prognostic resolution and reveal stage-specific vulnerabilities.

Artificial intelligence has transformed digital pathology by enabling biomarker prediction from high-resolution whole-slide images. However, current methods are computationally inefficient, processing thousands of redundant tiles per slide and requiring complex aggregation models. We introduce EAGLE (Efficient Approach for Guided Local Examination), a deep learning framework that emulates pathologists by selectively analyzing informative regions. EAGLE combines task-agnostic tile selection with detailed feature extraction and is benchmarked against leading slide- and tile-level foundation models across 43 tasks from nine cancer types spanning morphology, biomarker prediction, treatment response and prognosis. EAGLE outperforms patch aggregation methods by up to 23% and achieves the highest overall classification performance. It processes one slide in 2.27 s, reducing computational time by more than 99% compared with existing models. This efficiency supports rapid and auditable workflows by enabling review of the exact tiles used for each prediction and reducing dependence on high-performance computing. By reliably identifying informative regions and minimizing artifacts, EAGLE provides robust and auditable outputs, supported by systematic negative controls and attention concentration analyses. Its unified embedding enables rapid slide search, integration into multi-omics pipelines and emerging clinical foundation models.

The clinical translation of personalized cancer vaccines is currently impeded by the dual challenges of rapid antigen manufacturing and inefficient delivery to lymphoid tissues. In this study, we present a biohybrid nanovaccine platform that bridges synthetic biology and acoustic engineering to enable a noninvasive airway-to-lymph-node vaccination strategy. Leveraging an engineered nonpathogenic Escherichia coli TOP10 strain as living antigen factories, we construct ultrasound-responsive, manganese-chelated bacterial membrane nanovesicles that encapsulate biosynthesized tumor antigens. Following nebulized inhalation, low-intensity ultrasound triggers the acoustic droplet vaporization of these nanovesicles. This process induces cavitation-enhanced lymphatic trafficking, which actively overcomes pulmonary physiological barriers to promote deep tissue penetration and efficient antigen drainage to pulmonary lymph nodes. The spatiotemporally controlled co-delivery of antigenic proteins, innate bacterial adjuvants, and Mn2+ orchestrates a potent immune cascade, driving robust dendritic cell maturation and antigen cross-presentation. Consequently, this 'inhale-and-act' strategy elicits strong systemic antitumor immunity for next-generation personalized immunotherapy.

Aryl C-glycosides represent a valuable class of carbohydrates with broad biological activity in drug discovery, yet their rapid and stereoselective synthesis from unprotected native saccharides remains a significant challenge. Here we report a nickel-catalyzed, ligand-promoted stereoselective C1-arylation of unprotected saccharides with aryl bromides, enabled by glycosyl sulfonylhydrazide donors that are accessible in one step from native sugars. Notably, a series of N,N'-disulfonylpyridinedicarboxamidine tridentate ligands was developed and found to be critical for achieving both high reactivity and anomeric selectivity. The present method accommodates a broad range of aryl and heteroaryl bromides and is applicable to diverse mono- and oligosaccharides. To showcase the synthetic utility of this approach, we constructed a Taxol-sugar conjugate library. In vitro anticancer activity studies revealed that most conjugates retained significant antiproliferative activity against the tested cancer cell lines, while compound 6n was identified as selectively targeting glucose-dependent cancer cells with markedly improved water solubility compared with Taxol, emerging as a promising lead compound for cancer therapy.

Biomolecular condensates formed via phase separation are emerging targets for pharmacologic or genetic manipulation for cancer therapy. In this commentary, we envisage that further deciphering the composition and the physicochemical properties of oncogenic condensates will provide unprecedented opportunities to develop novel strategies for cancer chemotherapy and immunotherapy.

Oncogenic condensates act as biophysical sanctuaries that stabilize malignant survival programs. However, a universal regulator capable of orchestrating the integrated biophysical axes governing cellular phase behavior has remained elusive. Here, we introduce a sovereign singularity framework, presenting a deductive biophysical model that positions the indoleamine melatonin as a master regulator of biological phase separation. A systematic synthesis and integrative bioinformatics analysis were performed to identify the intersection between melatonin-responsive genes and the phase-separation proteome. We identified a core 26-gene regulatory signature-including AR, BCL2, CGAS, CTNNB1, EP300, EZH2, EGFR, IKBKG (NEMO), KEAP1, KDM1A (LSD1), LEF1, MYC, NANOG, PRNP (PRPc), SMAD3, SOX9, SQSTM1, TFEB, TFAM, TP53, TWIST1, USP10, WWTR1 (TAZ), VIM, YAP1, and YTHDF3-at the intersection of melatonin signaling and condensate architecture. We propose that melatonin utilizes a tri-lever framework of redox tuning (Lever I), multivalent plasticization (Lever II), and dielectric recalibration (Lever III) to render oncogenic programs biophysically untenable. This model provides a mechanical basis for high-resolution regulatory outcomes that modulate the organizational logic of nuclear decision-making (Axis I), state-transition (Axis II), and stress-adaptation (Axis III) condensates. Our results define a strategic platform for disrupting condensate-driven malignancy through the systemic modulation of the cellular biophysical landscape.

The complexity of cancer treatment calls for multimodal nanoplatforms integrating multiple therapeutic functions. Herein, we design a metal-ion-directed self-assembly strategy using a terpyridine-functionalized peptide (TyD) and demonstrate that coordination with different metal ions (Cu2+, Mn2+, Fe2+) simultaneously regulates nanostructure formation and dictates their enzyme-mimetic, DNA-binding, and photothermal properties. Notably, the TyD-Cu2+ complex emerged as a uniquely multifunctional platform, enabling efficient reactive oxygen species (ROS) generation, glutathione (GSH) depletion, DNA binding and cleavage, and exhibiting a high photothermal conversion efficiency (η = 33%). Relying on these integrated, metal-ion-dependent functionalities, we constructed a nanoplatform that synergistically combines chemodynamic therapy, photothermal therapy, and DNA-damaging interventions. This work establishes a metal-ion-tunable strategy for creating multifunctional peptide-based nanomaterials with potent antitumor effects in vitro, offering a promising material platform for further development in cancer therapy.

Biotin-avidin ligation represents the strongest known noncovalent protein-ligand interaction, serving as a cornerstone for high-affinity biosensing. However, the inherent tendency of biotin to promote nonspecific adsorption often compromises the sensitivity and reproducibility of such platforms. Silatranes have emerged as superior building blocks for surface functionalization of such biosensing platforms, offering enhanced hydrolytic stability, processability, and tunable silanization kinetics mediated by the internal transannular N→Si dative bond. In this work, we report the first synthesis of a biotinylated silatrane via azide-alkyne cycloaddition. The molecular structure was characterized using 1H NMR, FT-IR, and mass spectrometry. To develop a multifunctional biointerface, biotinylated silatrane was co-deposited with a zwitterionic sulfobetaine silatrane, creating a dual functional coating that integrates specific molecular recognition with potent antifouling properties. Surface characterization via water contact angle (WCA) goniometry, spectroscopic ellipsometry, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM) confirmed the formation of highly hydrophilic, uniform, and stable organosilicon adlayers. The specificity of the platform was validated through avidin bioconjugation, where quartz crystal microbalance with dissipation (QCM-D) and interfacial analysis confirmed the formation of a stable, well-ordered avidin layer. Finally, the biotinylated silatrane-based architecture was successfully employed for the detection of cancer biomarker carcinoembryonic antigen (CEA), demonstrating a linear response at clinically relevant concentrations. The findings establish biotinylated silatrane as a robust and versatile building block for the development of high-performance and fouling-resistant biointerfaces in diagnostic technologies and biosensing platforms.

Nuclei instance segmentation is critical in computational pathology for cancer diagnosis and prognosis. Recently, the Segment Anything Model has demonstrated exceptional performance in various segmentation tasks, leveraging its rich priors and powerful global context modeling capabilities derived from large-scale pre-training on natural images. However, directly applying SAM to the medical imaging domain faces significant limitations: it lacks sufficient perception of the local structural features that are crucial for nuclei segmentation, and full fine-tuning for downstream tasks requires substantial computational costs. To efficiently transfer SAM's robust prior knowledge to nuclei instance segmentation while supplementing its task-aware local perception, we propose a parameter-efficient fine-tuning framework, named Cooperative Fine-Grained Refinement of SAM, consisting of three core components: (1) a Multi-scale Adaptive Local-aware Adapter, which enables effective capability transfer by augmenting the frozen SAM backbone with minimal parameters and instilling a powerful perception of local structures through dynamically generated, multi-scale convolutional kernels; (2) a Hierarchical Modulated Fusion Module, which dynamically aggregates multi-level encoder features to preserve fine-grained spatial details; and (3) a Boundary-Guided Mask Refinement, which integrates multi-context boundary cues with semantic features through explicit supervision, producing a boundary-focused signal to refine initial mask predictions for sharper delineation. These three components work cooperatively to enhance local perception, preserve spatial details, and refine boundaries, enabling SAM to perform accurate nuclei instance segmentation directly. Extensive experiments on three challenging benchmarks demonstrate that our method achieves state-of-the-art performance, outperforming existing methods while requiring significantly fewer trainable parameters.

Magnetic resonance imaging (MRI)-guided radiotherapy (MRIgRT) integrates MRI with linear accelerators (MRI-linacs), enabling real-time motion management based on temporally resolved 2D MRI (cine-MRI). Current systems rely on template matching or deformable image registration for radiotherapy target (typically the gross tumor volume) localization, which allows beam gating. Further advances in localization could support more precise and efficient delivery methods. https://trackrad2025.grand-challenge.org/ was organized to provide a common dataset to benchmark algorithms for MRIgRT target tracking in 2D+t cine-MRI. Participants propagated target segmentation masks from an initialization frame across subsequent frames. The dataset comprised sagittal cine-MRI scans of 585 cancer patients undergoing radiotherapy at 0.35 T and 1.5 T MRI-linacs at six different institutions, with expert-annotated targets in 108 sequences. Target sites included the thorax (179 cases), abdomen (266 cases), and pelvis (140 cases). A total of 477 unlabeled and 50 labeled cases were provided for training purposes, 58 cases were kept private for preliminary testing (8) and final evaluation (50). The algorithms submitted by participants were executed on the challenge platform and assessed using metrics in three categories: geometric accuracy, surrogate dose accuracy and execution speed. Rankings were derived via a Rank-Then-Mean scheme. TrackRAD2025 attracted 148 registrations from 28 countries, 100 preliminary submissions and 24 final submissions from 14 teams. The top five methods achieved mean Dice similarity coefficients >0.87 and Euclidean center distances <2.1 mm, comparable to interobserver variability. Leading top five solutions featured foundation models with (4) or without (1) finetuning. Field strength had minimal effect on performance and tracking worked better for the pelvis with reduced motion amplitude compared to the thorax and abdomen cases, which achieved equivalent performance. TrackRAD2025 established a benchmark for MRIgRT tracking on multi-institutional cine-MRI data, highlighting foundation models as promising for clinical translation.

Oxidative stress arises from an imbalance between the production of reactive oxygen and nitrogen species (ROS and RNS, respectively) and the capacity of cellular antioxidant defenses to neutralize them. In cancer, this imbalance drives pathological remodeling of signaling networks that promote tumor initiation and progression. Among RNS, nitric oxide (·NO) and its highly reactive derivative peroxynitrite (ONOO-) are central mediators of redox dysregulation within the tumor microenvironment. These species induce site-specific post-translational modifications (PTMs), most notably protein tyrosine nitration, which can profoundly alter protein structure, function, interaction networks, and turnover, thereby reshaping essential cellular processes. In this review, we examine the molecular mechanisms of oxidative stress with a particular emphasis on nitration-driven protein modifications and their impact on oncogenic signaling. We highlight accumulating evidence that selective nitration of key signaling proteins actively contributes to multiple hallmarks of malignancy. These nitration events promote tumor initiation and growth, aberrant proliferation, migration and metastasis, metabolic reprogramming, angiogenesis, invasion, resistance to apoptosis, and immune evasion through disruption of core signaling pathways, cell-cycle control, and cell-death programs. Collectively, these findings support an emerging paradigm in which nitrated proteins are not merely passive byproducts of oxidative stress but active effectors of tumorigenesis. We discuss the translational implications of this concept, positioning protein tyrosine nitration as a source of mechanistically defined biomarkers and therapeutic vulnerabilities. A deeper understanding of the selectivity, structural consequences, and biological impact of protein tyrosine nitration will be essential for developing innovative precision strategies to modulate redox signaling in cancer and ultimately improve clinical outcomes.

Integrative analysis of complementary phenotype information from multi-modality data, such as pathological images and genomic profiles, has shown significant value in cancer survival prediction. However, multimodal survival prediction confronts two challenges: (1) the consistency and specificity of multimodal data remains underexplored, leading to incomplete information utilization and redundancy caused by overlapping information across modalities; (2) the correlation between inherent hierarchical structure of histopathological Whole Slide Images (WSIs) and genomic profiles has not been fully modeled. To address these issues, in this paper, we propose a Multi-mOdal survival prediction framework through Hierarchical Decoupling of whole-slide image pyramids and genomics (MoHD). Our MoHD incorporates adversarial information decomposition and hierarchical cross-modal interaction to advance survival prediction performance, which consists two core components: (i) a Multi-Granularity Feature Optimizer (MGFO) employing adversarial decoupling strategy to extract modality-common and refine modality-specific features while implementing redundancy suppression; (ii) a Multimodal Hierarchical INteractor (MHIN) that sufficiently captures multi-resolution cross-modal correlations and effectively integrates consistent and specific information through two scale-oriented interactors. We conduct extensive experiments on five cancer cohorts from the Cancer Genome Atlas (TCGA) database. The experimental results demonstrate that the proposed method achieves the superior performance compared to both unimodal and multi-modal survival prediction methods.

Paul Mischel, MD, and Alice Shaw, MD, PhD, the scientific program co-chairs for the American Association for Cancer Research Annual Meeting 2026, discuss some of the most exciting topics and key scientific themes to emerge from the meeting.

Despite revolutionizing oncology, cancer immunotherapy benefits only a minority of patients partly because of adaptive resistance. This resistance tends to manifest in two distinct clinical scenarios. The first is characterized by disease progression during treatment following an initial response, driven primarily by insufficient tumor-killing capacity. The second is characterized by relapse after initial remission, resulting from a failure to establish durable antitumor immune memory (AIM). Underlying both patterns is the progressive functional exhaustion of tumor-specific T cells, which reflects a dynamic equilibrium between immune attack and tumor evasion, a balance that is continuously challenged within an immunosuppressive tumor microenvironment (TME) sculpted by immunoediting. This review provides a comprehensive synthesis of current knowledge on adaptive resistance to cancer immunotherapy, with a particular emphasis on the mechanistic contributions of dysfunctional T-cell responses. We begin by outlining the frameworks of cancer immunoediting and AIM, detailing how exhausted T cells undermine sustained immunotherapeutic efficacy, and summarizing the achievements and limitations of current regimens. We then delineate how T-cell dysfunction, particularly within the TME and tumor-draining lymph nodes, culminates in adaptive resistance to immunotherapy, manifesting as either insufficient tumor-killing capacity leading to on-treatment progression or failure to establish durable AIM resulting in postremission relapse. A systematic analysis of the cellular and molecular drivers of this exhausted state follows, integrating established paradigms and highlighting key knowledge gaps. Finally, we critically evaluate emerging rejuvenation strategies, translational approaches for next-generation therapies, persistent challenges and promising directions for future research.

The thioredoxin (Trx) system, an integral component of cellular redox regulation, preserves protein dithiol-disulfide equilibrium through its conserved Cys-Gly-Pro-Cys active site and is involved in key cellular functions, including cell proliferation, apoptosis, and signal transduction. In cancer biology, the thioredoxin system plays dual roles: overexpression can suppress oxidative stress and promote tumor growth, whereas dysfunction can trigger programmed cell death (PCD). However, a critical area for future research is to delineate how Trx modulates the intricate networks of PCD, and to identify key nodes within these pathways that can be targeted for oncological therapy. This review outlines the structure and function of the thioredoxin system, highlighting its role in redox balance and its regulatory dynamics in healthy and disease states. We further examine the dual role of Trx by detailing its cross-regulatory networks that modulate diverse PCD pathways, including disulfidptosis, ferroptosis, apoptosis, autophagy, pyroptosis and necroptosis. In addition, we comprehensively outline therapeutic approaches that manipulate the Trx pathway to regulate PCD across a spectrum of health disorders, including malignancies, infectious diseases, neurodegenerative conditions, cardiovascular ailments, and metabolic dysfunctions. Finally, we address the current clinical applications of targeting the thioredoxin system. Although challenges such as tumor heterogeneity and drug-delivery efficiency persist, it remains a promising therapeutic avenue. This review aims to develop a theoretical framework and provide tactical guidance for the development of novel treatments targeting the Trx-PCD pathway.

Respiratory chain limitation is a recurrent but spatially heterogeneous state in solid tumors, driven by hypoxic subregions, electron transport chain defects, and signaling- or therapy-imposed respiratory inhibition. Restricted electron flow supports an ETC-linked MRS state characterized by accumulation of reduced electron carriers and configuration-dependent reactive oxygen species formation. This review describes how electron transport chain limitation remodels posttranslational modifications through (i) oxygen partitioning and substrate control of oxygen-dependent dioxygenases -considering that respiratory complex IV is the dominant intracellular molecular oxygen sink, thus shaping its availability for hydroxylation and demethylation reactions- and (ii) redox backpressure that shifts NAD(P)+/NAD(P)H balance, perturbs acyl-CoA and citric acid cycle metabolite pools, and rewires protonmotive force-linked matrix chemistry and thiol buffering. These constraints are predicted to remodel lysine acylations, HIF hydroxylation, histone and DNA methylation, cysteine-centered redox PTMs, phosphorylation networks, ubiquitin-dependent proteostasis, ADP-ribosylation, and lactylation. Together, these relationships support a site- and context-dependent PTM-routing state in which the position of respiratory chain limitation and the local tumor environment shape which PTM chemistries become rate-limiting, adaptive, or growth-restrictive.

Glutamate is the most abundant excitatory neurotransmitter in the central nervous system and a key non-essential amino acid in the body. It plays a central role in maintaining the excitatory-inhibitory balance of the nervous system, regulating systemic metabolic homeostasis, and participating in immune modulation. Additionally, it serves as a precursor for glutathione synthesis. Glutathione is a vital antioxidant and detoxifying molecule in organisms, capable of directly scavenging reactive oxygen species or toxic metabolites. Dysfunction of glutathione is associated with oxidative stress-related diseases. Given the critical roles of glutamate and glutathione in physiological and pathological states, we aim to systematically elucidates the metabolic interplay between glutamate and glutathione, referred to here as the Glutamate-Glutathione axis, as an integrative conceptual framework grounded in established biochemistry. By detailing the metabolic interplay of glutamate and glutathione, this review explores their mutually influential mechanisms. Furthermore, it summarizes the relevant signaling pathways and regulatory mechanisms within the Glutamate-Glutathione axis, clarifying its pivotal role in disease pathogenesis, with a particular focus on neuropsychiatric disorders and cancer. Finally, we aim to review non-invasive methods for detecting glutamate and glutathione using magnetic resonance imaging and presents our specialized sequence. This approach aims to provide a valuable non-invasive imaging tool for early diagnosis, efficacy evaluation, and therapeutic target development in related diseases. By integrating foundational metabolic networks, molecular regulatory mechanisms, disease associations, and clinical detection techniques, this review aim to establish the research framework of the Glutamate-Glutathione axis, fostering deeper integration and advancement from bench to bedside in this field.

Accurate cell counting in immunohistochemistry (IHC) images is critical for quantifying protein expression and aiding cancer diagnosis. However, the task remains challenging due to the chromogen overlap, variable biomarker staining, and diverse cellular morphologies. Regression-based counting methods offer advantages over detection-based ones in handling overlapped cells, yet rarely support end-to-end multi-class counting. Moreover, the potential of foundation models remains largely underexplored in this paradigm. To address these limitations, we propose a rank-aware agglomeration framework that selectively distills knowledge from multiple strong foundation models, leveraging their complementary representations to handle IHC heterogeneity and obtain a compact yet effective student model, CountIHC. Unlike prior task-agnostic agglomeration strategies that either treat all teachers equally or rely on feature similarity, we design a Rank-Aware Teacher Selecting (RATS) strategy that models global-to-local patch rankings to assess each teacher's inherent counting capacity and enable sample-wise teacher selection. For multi-class cell counting, we introduce a fine-tuning stage that reformulates the task as vision-language alignment. Discrete semantic anchors derived from structured text prompts encode both category and quantity information, guiding the regression of class-specific density maps and improving counting for overlapping cells. Extensive experiments demonstrate that CountIHC surpasses state-of-the-art methods across 12 IHC biomarkers and 5 tissue types, while exhibiting high agreement with pathologists' assessments. Its effectiveness on H&E-stained data further confirms the scalability of the proposed method. The code is publicly available at https://github.com/jtneuron/CountIHC.

Post-translational modifications (PTMs) are emerging as crucial regulators of proline metabolism, a pathway central to redox homeostasis, stress adaptation, and disease progression conserved across species. Beyond transcriptional regulation, PTMs such as phosphorylation, acetylation, and ubiquitination fine-tune the activity, stability, and localization of proline metabolic enzymes, including those involved in its biosynthesis and catabolism. Advances in proteomics and structural biology now provide insights into how these reversible modifications modulate enzyme oligomerization and metabolic flux, with involvement in plant stress tolerance and cancer cell survival. Here, we synthesize recent observations across kingdoms, discuss how PTMs integrate into metabolic control, and highlight future directions for exploiting PTM-based regulation in agriculture and human health.

Cardiovascular diseases are the leading cause of death worldwide, and cancer-associated thrombosis remains a major clinical challenge because of the interplay between tumour progression and platelet activation. Platelets contribute to thrombus formation by adhering to damaged endothelium and undergoing aggregation. Since mitochondria-targeted compounds are useful as antitumour and antiplatelet agents, we evaluated a series of triphenylphosphonium salts derived from gentisic acid alkyl esters with varying chain lengths, searching for antiplatelet agents with dual activity. The compound with a six-carbon chain (PSG6) exhibited the highest antiplatelet activity without increasing bleeding risk, whereas the cytotoxicity was found to increase with the chain length. PSG6 also showed selective anticancer effects, reducing tumour cell viability at micromolar concentrations, inducing mitochondrial fission, and lowering the mitochondrial membrane potential (ΔΨm) at 10 μM. Mechanistically, PSG6 decreased ΔΨm, inhibited the mitochondrial electron transport chain (ETC) at complex I, and increased intracellular calcium and reactive oxygen species (ROS) production. Complex I inhibition was confirmed in the yeast model organism Yarrowia lipolytica (IC50 = 2.9 μM), and atomistic molecular dynamics simulations suggest that PSG6 may inhibit complex I by binding at the shallow site of the ∼30 Å long ubiquinone tunnel. These results position PSG6 as a promising mito-inhibitor candidate for antiplatelet therapy.

Telomere crisis contributes to cancer genome evolution. Beyond the loss of end protection, replication defects at short telomeres give rise to aberrant fork intermediates that can be resolved by microhomology-mediated end joining. Such mutagenic repair yields chromosomal fusions and complex rearrangements that shape cancer genomes.

Sirtuin 2 (SIRT2) is an NAD⁺-dependent lysine deacylase that is a member of the sirtuin enzyme family and plays essential roles in cytoskeletal regulation, chromatin remodeling, metabolic control, inflammation, neurodegeneration, and cancer progression. Its diverse biological functions have positioned SIRT2 as a compelling but challenging therapeutic target. This review provides an integrated overview of recent advances in SIRT2 structural biology, emphasizing the catalytic core, substrate-binding channel, and the inducible selectivity pocket that enables isoform discrimination. We summarize the medicinal chemistry landscape of classical SIRT2 inhibitors, highlighting major scaffolds and determinants of potency and selectivity. Emerging strategies based on targeted protein degradation-including SirReal-derived PROTACs, hydrophobic-tag degraders, and non-CRBN E3 ligase systems-are discussed in comparison with traditional occupancy-driven inhibition, underscoring the advantages of event-driven degradation for eliminating both catalytic and non-catalytic SIRT2 functions. Drug repurposing efforts and computational screening approaches further expand the repertoire of potential SIRT2 modulators. Finally, we outline current challenges and future directions, including the need for improved selectivity, better pharmacokinetic profiles, deeper mechanistic understanding, and development of chemical probes for underexplored sirtuin isoforms. Together, these advances highlight the rapidly evolving landscape of SIRT2-targeted therapeutics and their emerging potential in oncology, neurodegeneration, and metabolic disease.

Okara, a byproduct of soybean processing, can be valorized through microbial fermentation to yield bioactive compounds. In this study, we used Lacticaseibacillus paracasei, a Gram-positive lactic acid bacterium commonly applied in probiotic and dairy fermentations, to perform solid-state fermentation of okara. Crude polysaccharides were then purified using DEAE Sepharose Fast Flow and BioGel P-2 gel filtration chromatography, yielding a high-purity β-1,4-galactan polysaccharide fraction (LPP-A) with an average molecular weight of 6 kDa. To assess its immunostimulatory potential, LPP-A was evaluated as an adjuvant in a murine vaccination model with ovalbumin (OVA) as the antigen. Mice immunized with OVA plus LPP-A induced robust OVA-specific cytotoxic T lymphocyte (CTL) responses and high IgG antibody titers, indicating the induction of strong antigen-specific cellular and humoral responses. In a subsequent tumor challenge, the OVA plus LPP-A group showed significantly inhibited EG7-OVA tumor growth, suggesting high potential for LPP-A as an immune-enhancing adjuvant for cancer immunotherapy. These results demonstrate that fermenting okara can produce a β-1,4-galactan (LPP-A) with potent vaccine adjuvant properties, supporting its value in protein-based immunization strategies.

Redox metabolism plays a pivotal role in cancer initiation, progression, and resistance, making it an attractive target for intervention. Nanoscale metal-organic frameworks (MOFs) have emerged as versatile platforms for redox modulation owing to their intrinsic redox activity and high porosity for therapeutic encapsulation. Their modular architectures allow the integration of redox-active metal nodes and functional ligands to directly influence redox processes, while the porous frameworks facilitate efficient loading and controlled release of therapeutic agents. This review summarizes recent advances in MOF-based redox modulation strategies, including approaches to increase reactive oxygen species accumulation via external stimuli or catalytic pathways, suppress antioxidant systems such as glutathione, thioredoxin, and NAD(P)H, and modulate redox-related signaling pathways that govern tumor metabolism and immunity. We conclude with a discussion of key challenges, including biosafety, stability, and clinical translation, and highlight opportunities for future research at the interface of MOF-based nanomedicine and tumor redox biology.

Medical image segmentation is challenging due to the diversity of medical images and the lack of labeled data, which motivates recent developments in federated semi-supervised learning (FSSL) to leverage a large amount of unlabeled data from multiple centers for model training without sharing raw data. However, what remains under-explored in FSSL is the domain shift problem which may cause suboptimal model aggregation and low effectiveness of the utilization of unlabeled data, eventually leading to unsatisfactory performance in unseen domains. In this paper, we explore this previously ignored scenario, namely domain generalized federated semi-supervised learning (FedSemiDG), which aims to learn a model in a distributed manner from multiple domains with limited labeled data and abundant unlabeled data such that the model can generalize well to unseen domains. We present a novel framework, Federated Generalization-Aware Semi-Supervised Learning (FGASL), to address the challenges in FedSemiDG by effectively tackling critical issues at both global and local levels. In our proposed framework, globally, we introduce Generalization-Aware Aggregation (GAA), assigning adaptive weights to local models based on their generalization performance. Locally, we use a Dual-Teacher Adaptive Pseudo Label Refinement (DR) strategy to combine global and domain-specific knowledge, generating more reliable pseudo labels. Additionally, Perturbation-Invariant Alignment (PIA) enforces feature consistency under perturbations, promoting domain-invariant learning. Extensive experiments on four medical segmentation tasks (cardiac MRI, spine MRI, bladder cancer MRI and colorectal polyp) demonstrate that our method significantly outperforms state-of-the-art FSSL and domain generalization approaches, achieving robust generalization on unseen domains. This work provides a practical solution for addressing domain shifts in federated semi-supervised learning, advancing multi-center collaboration in privacy-sensitive healthcare applications. The code will be made public upon acceptance.

Cells undergoing transitional states have been broadly referred to as plastic intermediates emerging between stable identities in multiple biological contexts. Once regarded as indistinct midpoints on lineage trajectories, these states are now recognized as discrete, biologically meaningful epigenetically permissive states, exquisitely responsive to environmental and stress signals at critical junctures of biological events that confer competence to proceed along their trajectories. These high-plasticity nodes have emerged as central regulators of developmental progression and determinants of disease outcomes, serving as functional bottlenecks in which resolution or persistence dictates normal or maladaptive pathological responses. Recent single-cell and multiomics technologies enabled their detection with unprecedented resolution, revealing conserved regulatory themes, including stress-response activation and striking context dependence shaped by niche cues and tissue architecture. Yet challenges remain in capturing their rapid heterogeneous dynamic in the multiple contexts, and defining their function, in vivo. Here we summarize current concepts on the identification, diversity, role, and regulation of these cell states in events from early development to adult homeostasis, repair, and disease. The increasing recognition that transitional states can be productive conduits or pathological traps underscores their relevance in these processes and potential for the identification of therapeutic targets for intervention in disease, cancer, and regenerative medicine.

Accurate and automated whole slide image (WSI) classification is important in early diagnosis of cancer, which is typically achieved using multiple instance learning (MIL). Existing MIL approaches for WSI classification still face practical challenges, particularly related to large numbers of patches cropped from a WSI with only slide-level labels. In these cases, training often suffers from overfitting due to the weak supervision provided by the slide-level labels. Therefore, it is crucial to exploit more information from limited slide-level data for WSI analysis. However, existing approaches concentrate on only single-magnification feature mining, which fails to capture feature consistency among different magnifications. To address this shortcoming, we here propose a multiple-magnification optimal transport-based cross-attention (M2OTCA) MIL framework for WSI classification, in which optimal transport (OT) is applied to match feature distributions of a WSI across different magnifications for selecting magnification-specific informative patches. To alleviate the high computational complexity, we effectively summarize the magnification-specific basic structural distribution at each magnification by condensing constituting instances as region structural prototypes, and modeling the original OT learning as cross-magnification region structural prototypes OT learning. Moreover, to generate a faithful explanation for cross-magnification OT learning, we derive region structural prototype gradients at the high magnification prototypes (20×) as the indicators to enhance low magnification prototypes (10× and 5×) for efficient feature co-expressions. Then, prototypes at each magnification are aggregated for magnification-specific predictions which are concatenated for final prediction. Extensive experiments on four representative WSI datasets show the superiority of M2OTCA compared to other state-of-the-art methods.

Controlled release delivery systems have become a focal point in the pharmaceutical sector, with biopolymers playing a crucial role in enabling precise modulation of active compound release. Polymers, categorized into natural and synthetic types, are widely used due to their biocompatibility, biodegradability, and versatility. Natural biopolymers, such as proteins, polysaccharides, and microbial polyesters, offer high bioactivity but may encounter challenges related to mechanical strength and degradation rates. Synthetic polymers, including poly(lactic acid) and polycaprolactone, are engineered to provide enhanced stability and tunable release profiles. Integrating these biopolymers in controlled release systems enhances targeted therapeutic outcomes by ensuring sustained drug release in pharmaceutical applications, thereby improving drug efficacy and minimising side effects. This review demonstrates that utilizing novel approaches involving biopolymeric materials for controlled-release delivery systems is a promising route for precise control, offering important industrial potential. Integrating these materials in the pharmaceutical industry represents a breakthrough in enhancing release profiles and targeting specific applications (e.g. cancer therapy, drug delivery, nutraceutical delivery). The latest advancements in biopolymeric materials and their applications in controlled release systems, focusing on targeted release strategies, are comprehensively discussed. Key release mechanisms (diffusion, erosion, and swelling) are also critically compared for targeted applications. Despite progress, challenges like scalability and in vivo validation require further research to bridge lab-scale innovations to commercial applications. Biopolymeric delivery systems enable controllable release kinetics through material-structure interactions, supporting rational design of targeted pharmaceutical formulations with translational relevance.

Brain metastasis remains a devastating disease with dismal prognosis. How circulating tumor cells (CTC) penetrate the blood-brain barrier (BBB) and reprogram the brain microenvironment remains unclear. Using spatially resolved multi-omics profiling of CTCs and brain metastases, integrated with experimental and clinical analyses, we identified glycoprotein nonmetastatic melanoma protein B (GPNMB) as a CTC-secreted driver of vascular disruption and brain colonization. CBX3 upregulation induced GPNMB expression, which bound endothelial EGFR, triggering CBL-mediated ubiquitination and degradation. Attenuated EGFR signaling suppressed FTO and disrupted endothelial junctions via YTHDF2-dependent TJP1 m6A methylation. Remarkably, GPNMB-induced BBB remodeling promoted immune infiltration via the CXCL12-CXCR4 axis and induced time-course-dependent T-cell exhaustion within the brain microenvironment. Clinically, elevated CBX3+GPNMB+ CTCs and plasma CXCL12 were significantly associated with brain metastasis progression in lung cancer and melanoma. Therapeutically, dual blockade of GPNMB and PD1 enhanced anti-brain metastasis efficacy in mice, unveiling GPNMB as a promising target for precision immunotherapy. GPNMB is a CTC-secreted driver of BBB disruption and brain colonization via the CBX3-GPNMB-EGFR-FTO-TJP1 axis. GPNMB-induced BBB remodeling promotes CXCL12-CXCR4-mediated immune infiltration and enhances T-cell exhaustion, sensitizing brain metastasis tumors to GPNMB/PD1 dual blockade. CBX3+GPNMB+ CTCs and plasma CXCL12 may serve as noninvasive biomarkers for brain metastasis management.

Ferroptosis is a cell death process defined by the iron-mediated peroxidation of membrane phospholipids that overwhelms the cell's innate antioxidant capabilities. Sitting at the nexus of iron, lipid, reactive oxygen species stress responses, and cellular metabolism, ferroptosis is intricately tied to these pathways. The burgeoning field of cancer metabolism has revealed that cancer cells exhibit changes in ferroptosis-relevant metabolic pathways, thereby opening an important avenue of investigation into whether tumors can have characteristic metabolic alterations that render them exquisitely sensitive to ferroptotic cell death. In this review, we highlight recent findings in the metabolic pathways linking ferroptosis and oncogenesis, as well as implications for future cancer therapeutic strategies.

Iron-based chemodynamic therapy (CDT) is an emerging cancer treatment strategy that utilizes the overexpression of hydrogen peroxide (H2O2) and mild acidity within the tumor microenvironment. This approach generates highly toxic hydroxyl radicals (·OH) within tumors through iron-ion-catalyzed Fenton or Fenton-like reactions, enabling selective tumor cell ablation while minimizing damage to healthy tissues. Iron is recognized as an ideal catalytic metal for CDT due to its excellent biocompatibility, tumor-targeting potential, and versatile theragnostic capabilities. This review explains the core mechanisms of iron-based CDT, analyzes the inherent limitations posed by the tumor microenvironments, and comprehensively summarizes advanced iron-based nanomaterials designed to overcome these challenges, including inorganic nano catalysts, organic-inorganic hybrids, and biomimetic nanostructures. Furthermore, it highlights multidimensional strategies to enhance CDT efficacy: actively remodeling the tumor microenvironment by modulating H2O2 supply, pH, antioxidant defenses and hypoxia; using external stimuli such as light, ultrasound, or magnetic fields to remotely activate and amplify reactions; and achieving synergistic effects by combining with chemotherapy, radiotherapy, and immunotherapy. Finally, the review outlines key challenges in the clinical translation of iron-based CDT and suggests future research directions, providing a comprehensive and timely perspective on this rapidly advancing field.

Development of de novo therapeutic agents is a complex, costly, and a high-risk process, whereas repurposing approved and investigational drugs for novel targets offers a more efficient and cost-effective strategy, likely yielding higher success rates. This approach demonstrated its effectiveness during SARS-CoV-2 pandemic, when existing drugs were repurposed to combat the virus. Originally pivotal in developing antiviral treatments against HIV and HCV, viral protease inhibitors represent a structurally privileged class of compounds capable of targeting difficult and non-classical protein sites. They have demonstrated promising biological activity against diverse alternative targets, including fungal pathogens, multidrug-resistant bacteria, and cancer, making them prime candidates for repurposing. This mini-review highlights the unique structural and physicochemical properties of approved HCV and HIV viral protease inhibitors that enable their repurposing for the development of new therapeutic agents.

CRISPR-Cas9 genome-wide screening has been instrumental towards identifying novel targets for drug discovery in cancer research. However, much of this research has centred specifically on adult cancers, with paediatric cancers being underserviced by current research and screening. With contemporary evidence increasingly highlighting the differences in biology between adult and children's cancers, more research has gradually occurred investigating dependencies and mechanisms of resistance in paediatric cancers through the use of CRISPR-Cas9 genome-wide screens. This review collates and summarises the experimental genome-wide screens that have been performed specifically in paediatric cancer models, and highlights the versatility of this technology and the knowledge gained through this research.

Immunotherapy has revolutionized the treatment of cancers, infections, and autoimmune disorders by harnessing immune mechanisms such as signal recognition, intracellular cascades, and mediator release. However, clinical translation remains hindered by poor specificity, dysregulated signaling, and heterogeneous responses, leading to toxicity and resistance. Synthetic biology-engineering programmable cellular systems-provides transformative strategies to overcome these barriers. Through designed receptors, modular signaling circuits, and precision-controlled immune behaviors, it enables next-generation therapies with enhanced specificity and tunability. Key advances include synthetic immune circuits that permit context-dependent functions such as logic-gated activation and feedback-controlled cytokine secretion. These innovations open avenues for personalized treatments, bridging bioengineering with immunology to maximize efficacy and minimize off-target effects. This review highlights the translational potential of synthetic biology in advancing precision immunotherapy for oncology and immune disorders.

Vessel-microenvironment interactions are pivotal in tissue development and disease. Existing experimental platforms fail to recapitulate the seamless extracellular matrix (ECM) continuity required for physiological vessel-stromal communications, often creating disruptive material boundaries. In this study, we developed a consubstantial bioink-supporting bath system using collagen modified with nanoscale PEGylated polyhedral oligomeric silsesquioxane, enabling interface-free vessel-microenvironment modeling. This suspension matrix approach maintains uninterrupted, biomimetic ECM to preserve spatial complexity and multicellular crosstalk. We validated this platform through complementary models: a regenerative vessel-stromal model demonstrating the dominance of contact-dependent signaling over paracrine effects and a pathological vessel-tumor model. Single-cell RNA sequencing and cell-cell communication analysis revealed cancer-associated fibroblast-mediated networks driving pathological angiogenesis. This study advances vessel-microenvironment biology and provides a platform to investigate vascular mechanisms and therapies.

Antibody-drug conjugates (ADCs) are a leading area of targeted cancer therapeutics, typically combining a tumor-associated antigen-specific antibody conjugated to a toxic payload that targets key cellular mechanisms, such as mitosis and survival. The global ADC clinical trial landscape has been expanding significantly, with over 430 ADCs reaching early to late clinical studies in the past two decades, up from just 90 between 2004 and 2014. The US Food and Drug Administration (FDA) has so far approved 14 ADCs for use in clinical oncology. This growth is likely driven by significant advances in antibody technology and conjugation methods enabling more effective and precise delivery to cancer cells and more effective payloads that target vital cancer biology. Here, we review the ADCs that have reached clinical approval as well as current and emerging trends in ADC development, and we discuss these from multiple perspectives, including ADC mechanisms of action, emerging antigen targets, linker and conjugation chemistry, payloads, combination of ADC with checkpoint inhibitor immunotherapy, and antibody fragment crystallizable (Fc) engineering. We also consider how the field is evolving through the application of artificial intelligence (AI) and pathology-based biomarker discovery. Combined, innovative and emerging ADC design coupled with precision medicine and patient stratification strategies hold great promise to develop diverse and personalized cancer treatments with improved therapeutic indices and to enhance tolerability compared to traditional chemotherapy and current established ADCs. This review aims to assist researchers in exploring the evolution, characteristics, and development trends in ADC design and to provide new directions for future research.

The activity and distribution of Piezo1 molecules, along with the maturity and strength of focal adhesions (FAs), serve as critical factors influencing cell mechanosensing. Notably, migrating epithelial cells and mesenchymal-like cancer cells exhibit significantly different behaviors regarding these elements. In cancer cells, Piezo1 molecules are distributed uniformly, while in epithelial cells, their distribution is heterogeneous. In epithelial cells, Piezo1 molecules tend to group around FAs, a phenomenon that is enhanced by actomyosin contractility. However, a reduction in contractility results in a more uniform distribution of Piezo1 molecules. The expression and activity levels of Piezo1 molecules are markedly higher in cancer cells compared to epithelial cells. The activity of Piezo1 molecules correlates with the intracellular calcium concentration. Despite the extensive experimental studies on the properties of migrating epithelial and mesenchymal-like cancer cells, the physical explanations remain lacking. The primary objective of this theoretical study is to explore: (i) the inhomogeneous distribution of Piezo1 molecules in epithelial cells in relation to the Marangoni effect, (ii) the heightened activity of Piezo1 molecules in cancer cells by specifying the driving force, and (iii) the influence of membrane-mediated interactions among Piezo1 molecules grouped near FAs in epithelial cells on their activity.

Activation of unfolded protein response (UPR) has recently been identified as a new target for cancer therapy. However, pharmacological agents activating UPR have not been approved in clinical practice. In this study, spatiotemporally controllable UPR activation was realized by the combination of ultrasound (US)-triggered sonodynamic therapy (SDT) with titanium nitride (TiN) NPs and proteasome inhibitor bortezomib (BTZ). BTZ-conjugated TiN NPs were prepared by using a linker of carminic acid (CA)-functionalized bovine serum albumin (BSA). CA interacts with BTZ to form dynamic boronate ester bond enabling pH- and ultrasound-responsive drug release and thus overcoming the challenge of poor drug release for the conventional delivery systems. Compared with traditional sonosensitizer TiO2, TiN has superior SDT efficiency. TiN-mediated SDT inhibits not only the β1/β5 active sites of proteasomes, but also the β2 proteasome activity not targeted by BTZ, thus synergistically triggering endoplasmic reticulum (ER) stress. Moreover, SDT and BTZ complement each other perfectly to activate all three pro-apoptotic UPR pathways including PERK/eIF2α/ATF4, IRE1/JNK and ATF6, thereby leading to dramatic upregulation of CHOP expression, calcium overload and mitochondrial dysfunction. Importantly, this intensified ER stress effectively triggers immunogenic cell death (ICD), characterized by the massive release of damage-associated molecular patterns (DAMPs), thereby further recruiting and activating cytotoxic T lymphocytes and reshaping the immunosuppressive tumor microenvironment. In summary, making up the current lack of effective pharmacological agents activating UPR, this study provides a new strategy to spatiotemporally activate UPR through the ingenious combination of SDT and proteasome inhibition with BTZ-conjugated TiN NPs.

Assays for cancer diagnosis via the analysis of tumor biomarkers on circulating extracellular vesicles (EVs) have shown great potential. Single EV imaging that can measure the abundance of protein biomarkers in EVs can help in detecting the presence, stage, and progression of disease. Antibodies are typically used to detect EV proteins. Controlling the quality of antibody-based immunoassays can, however, be challenging, as they may exhibit unintended cross-reactivity with non-target proteins or variability in binding affinity across different batches, even for monoclonal antibodies. Here, we report short peptides that are a promising alternative to antibodies for detecting protein biomarkers in EVs. We describe a pipeline that combines a Peptide Binding Design (PepBD) algorithm and molecular-level simulations to identify peptides that can recognize the extracellular domain of EpCAM (a known cancer biomarker). The peptides designed for EpCAM were labeled with a fluorescent dye and their binding to the target protein was evaluated using fluorescence ELISA. The results identified a computationally designed peptide that exhibited an affinity for the target EpCAM protein comparable to that of an antibody. Further testing involved single EV imaging to gauge the peptide's affinity towards EVs.

Over the past decade, cancer vaccines have shown great promise in immunotherapy for solid tumors. However, the efficacy of cancer vaccines is unsatisfactory due to the complexity, heterogeneity, and immune evasion of cancer, as well as the instability of the core components of the vaccines (antigens and adjuvants), the weak immunogenicity and low presentation efficiency of antigens, and the inability to effectively activate immune cells. Nanotechnology is considered to be a transformative approach to address these challenges by improving vaccine delivery. As carriers and/or adjuvants, nanoparticles take advantage of their superior physicochemical properties to enhance the stability of antigens and adjuvants, achieve controlled release in time and space, enable flexible and synergistic combination therapies, and create highly targeted delivery systems, thereby optimizing the efficacy and durability of antitumor immunity and minimizing side effects. In this review, the key components of the nanovaccine strategy are highlighted, which covers antigen forms, adjuvant types, nanovaccine platforms, and more. We provide the latest advances in the development of cutting-edge biomaterials and carrier systems for controlled vaccine delivery. Finally, integrating the current progress strategy, we offer critical perspectives on future applications of nanomaterials in cancer vaccines for clinical translation.

Chimeric antigen receptor (CAR) therapy has transformed the treatment landscape for hematologic malignancies, but its efficacy in solid tumors is limited, owing in part to insufficient functional persistence of the engineered T cells. To elucidate the basis for their functional decline, we conducted integrated chronic in vivo and in vitro screens of 400 transcription factors, which revealed NFIL3 as a driver of CAR T-cell dysfunction. Genetic disruption of NFIL3 in CAR T cells sustains their expansion and increases cytokine production, overall restraining terminal differentiation. Loss of NFIL3 enhances CAR T-cell efficacy, improving tumor control and prolonging survival in xenograft and syngeneic mouse tumor models across different CAR designs. Under chronic stimulation, disruption of NFIL3 establishes a transcriptional state predictive of favorable clinical outcomes. Our findings underscore the power of comprehensive in vivo genetic screens integrated with multiparameter in vitro assessment and identify NFIL3 as a novel therapeutic target to enhance cancer immunotherapy. This study presents a two-step screening framework, integrating an in vivo pooled guide RNA screen with a multiparameter, in vitro arrayed screen. NFIL3 emerged as the top candidate, and its disruption enhanced CAR T-cell antitumor efficacy in both hematologic malignancies and solid tumors across diverse CAR architectures.

Ferroptosis is an iron-dependent cell death characterized by the disorder of iron homeostasis and the accumulation of lipid peroxidation, which trigger a cascade of events resulting in cell death. This process has a dual regulatory effect in the evolution of diseases, which can not only attenuate the progress of diseases, but also become a promoting force to accelerate pathological alterations, and widely participate in the occurrence and development of multiple diseases. In recent years, there has been a growing body of research identifying regulators-both inducers and inhibitors-that target the mechanisms of ferroptosis, particularly in the field of anti-tumor drug research. This review systematically elucidates the regulatory mechanisms underlying ferroptosis (e.g., iron metabolism, lipid peroxidation, system Xc-/GSH/GPX4 axis) and examines the role of ferroptosis in the pathogenesis of various diseases, such as cancer, neurodegenerative diseases, liver and kidney diseases, and cardiovascular diseases. Additionally, we discussed the therapeutic applications of novel inducers and inhibitors of ferroptosis in recent years, offering critical insights and directions for further researching on the mechanism of ferroptosis and targeted drugs for ferroptosis-related diseases.

Sonocatalytic therapy has emerged for efficient malignant tumor treatment due to its non-invasive characteristics, spatiotemporal precision, and deep tissue penetration capabilities. The defect engineering can improve charge carrier separation, and enhance reactive oxygen species (ROS) generation, but the current technological limitations in developing efficient hole trapping sites relative to electron traps still impose significant constraints on overall ROS production efficiency. In this work, the rational design of two-dimensional Fe-vacancy (FeV)-engineered FeOCl nanosheets (FeVOCl NSs) with iron vacancies enables simultaneous electron and hole (e--h+) trapping, which confers not only significantly enhanced sonocatalytic activity but also multienzyme-mimicking properties, including peroxidase-, oxidase-, and catalase-like activities. These capabilities collectively promote ROS generation, relieve tumor hypoxia, and reshape the tumor microenvironment. Density functional theory (DFT) calculations reveal that strategically engineered iron vacancies in FeVOCl NSs significantly improve ROS generation efficiency through facilitating charge carrier separation and increasing H2O2 adsorption capacity. This dual functionality under ultrasound (US) irradiation leads to efficient ROS-mediated activation of both apoptotic and pyroptotic cell death pathways in tumor cells, which achieve significant in vivo sonocatalytic tumor-eliminating capacity and efficacy across multiple cancer models, including hematologic and breast cancer. These findings highlight the essential role of strategic defect engineering coupled with enzyme-mimicking activity in optimizing sonocatalytic therapeutic outcomes for efficient cancer treatment.

Adenosine triphosphate (ATP) is the central energy currency in cells, driving essential processes such as biosynthesis, transport, motility, and signal transduction. The dysregulation of ATP homeostasis is closely linked to many pathological conditions, including cancer (through the Warburg effect and extracellular ATP signaling), neurodegenerative diseases characterized by mitochondrial dysfunction, ischemia-induced cell death, and impaired tissue regeneration. Among the strategies being explored in their treatment, ATP-responsive and energy-regulating biomaterials have emerged as innovative platforms for precisely modulating pathological microenvironments and cellular metabolism. This review examines ATP-modulating biomaterials by categorizing them into four major classes: ATP-responsive materials, energy-conversion materials, ATP-functionalized materials and ATP-detection materials. It summarizes the mechanisms underlying dynamic ATP fluctuations in various diseases and their biomedical implications, with a focus on oncology, antibacterial therapy, neurodegenerative disorders, and tissue repair. By integrating perspectives from materials science, nanotechnology, and biomedicine, the review highlights key challenges, such as biocompatibility, specificity, and translational barriers, while proposing future research directions to advance ATP-based biomaterials toward improved therapeutic outcomes.

Mitochondria are essential for cell energy metabolism, redox homeostasis, and apoptosis. Meanwhile, numerous pathological conditions are linked with mitochondrial dysfunction, such as cardiovascular diseases, connective tissue disorders, chronic wounds, neurological disorders, and cancer. Mitochondria-targeted hydrogels (MTHs) have emerged for their ability to selectively deliver active agents to mitochondria, modulate mitochondrial function precisely, which plays a key role in improving treatment efficacy. Hydrogels offer unique advantages, including biocompatibility, structural tunability, and controlled drug release. With specific targeting ligands and stimuli-responsive mechanisms, hydrogels can achieve mitochondrial localization and therapeutic modulation. Recent advancements have demonstrated significant benefits of MTHs in reducing oxidative stress, promoting oxidative phosphorylation and restoring mitochondrial quality control across a variety of disease models. However, challenges remain, including optimizing targeting efficiency, as well as accuracy. Further exploration of therapeutic mechanisms and the integration of multi-dimensional targeting strategies are also essential for the clinical application of MTHs. The current review highlights the development of mitochondrial targeting strategies and specifically focuses on a series of applications of MTHs in mitochondrial-related diseases. Lastly, the discussion delves into the shortcomings of existing therapies and possible future research ideas.

Cuproptosis represents a promising avenue for cancer immunotherapy, yet its efficacy is limited by difficulties in achieving targeted mitochondrial copper ion accumulation and upregulating cells reliance on mitochondrial respiration. To address this, we develop a biomimetic copper-based nanoplatform (SCTDM) coated with macrophage-tumor cell hybrid membranes (HM) and co-loaded with the lactate modulator syrosingopine (Su3118) and the copper ionophore disulfiram (DSF). SCTDM integrates active tumor targeting, in situ photothermal therapy (PTT), and lactate metabolism modulation, synergistically enhancing cuproptosis and antitumor immunity. SCTDM releases copper ions and DSF to facilitate mitochondrial copper ions delivery and induce immunogenic cell death (ICD). In the high hydrogen sulfide (H2S) tumor microenvironment (TME), copper ions further form photothermal copper sulfide complexes that enhance PTT efficacy and reactive oxygen species (ROS) generation, thereby augmenting cuproptosis. Notably, the liberated Su3118 selectively inhibits monocarboxylate transporters (MCT1/4), suppressing lactate efflux while simultaneously sensitizing cells to cuproptosis by disrupting glycolysis and restoring mitochondrial respiration. Collectively, the synergistic combination of PTT and lactate metabolism modulation serves to potentiate cuproptosis-induced immunogenic cell death. This cascade effect culminates in the reprogramming of the immunosuppressive TME and augments anti-tumor immunity for eradicating primary tumors and preventing recurrence.

Over the last 25 years, antibody-drug conjugates (ADCs) have evolved from a novelty to a well-established and clinically validated tool for cancer treatment. Most ADCs are prepared by conjugating one payload per site via linear linkers. However, when conjugation is performed at a single position or a higher drug-to-antibody ratio (DAR) is desired, it becomes necessary to conjugate multiple drug moieties per site. This requires the use of multivalent linkers, which can be either low-molecular-weight branched structures or polymers. Several ADCs based on these linkers have reached clinical trials, and branched linkers are now being actively explored for preparing highly promising dual-payload conjugates. In this review, we comprehensively examine various multivalent linker architectures and explore the interplay between linker-payload design, conjugation strategies, and ADC properties and activity. We also evaluate the unique challenges associated with multivalent ADC linkers, such as increased local hydrophobicity and hindered payload release, as well as strategies for overcoming these hurdles. Further, we provide a future outlook for multivalent linker technologies, considering current trends in the rapidly evolving field of ADC development.

Lymph node metastasis is a crucial factor in cancer prognosis; however, no clinically approved contrast agents are available for its accurate detection via medical imaging. The contribution of macrophage uptake to nanoparticle (NP) accumulation during lymphatic drainage has been a subject of ongoing debate. Herein, we report a novel lymph node imaging strategy designed to enhance lymph node retention while evading macrophage clearance through nanoprobe surface modification, thereby improving the diagnostic accuracy of lymph node metastases. We engineered a series of Pluronic F127-based NPs as magnetic resonance imaging (MRI) contrast agents, incorporating manganese (Mn) porphyrin molecules bearing varied functional groups (-CH3, -OH, -COOH, -NH2, and -B(OH)2). The influence of these surface modifications on physicochemical properties and macrophage-evading capabilities was investigated. Notably, boronic acid-functionalized Mn-porphyrin NPs exhibited exceptional macrophage evasion, demonstrating significantly reduced macrophage uptake in vitro and prolonged blood circulation in vivo relative to NPs modified with alternative functional groups. In a lymph node metastasis model, boronic acid-modified Mn-porphyrin NPs administered via local footpad injection (0.005 mmol Mn/kg) achieved optimal lymph node MRI contrast with an extended imaging window. Furthermore, in New Zealand rabbits, even at lower doses (0.0025 mmol Mn/kg), boronic acid-modified Mn-porphyrin NPs provided excellent lymphatic system visualization, highlighting the potential for further translational studies. In summary, this study confirms the feasibility of a boronic acid group modification strategy that evades macrophage uptake and utilizes lymphatic drainage to diagnose lymph node metastasis, offering valuable insights for the development of next-generation macrophage-evading nanoprobes.

The rat sarcoma virus oncogene (RAS) is one of the most frequently mutated drivers in human cancers. Developing targeted therapies against RAS has been challenging due to its structure. The recent breakthroughs with covalent Kirsten RAS (KRAS) inhibitors represent a key milestone in targeting mutant KRAS proteins. However, drug resistance remains a significant obstacle. The proteolysis-targeting chimeras (PROTACs), which degrade mutant KRAS proteins, offer a promising strategy to overcome resistance and expand therapeutic options. This review covers the structural basis of KRAS and signaling networks, while discussing recent advancements in KRAS inhibitor research and PROTAC technology. It aims to provide a foundation and inspiration for future KRAS inhibitor and degrader development.

Cold atmospheric plasma (CAP) has emerged as a promising non-thermal modality in cancer research due to its ability to induce selective cytotoxicity through reactive oxygen and nitrogen species. However, the limited penetration depth and instability of plasma-derived reactive species in complex biological environments remain major obstacles to its therapeutic application. In this study, we investigated whether exosomes derived from CAP-treated cancer cells (CAP-Exo) could serve as functional mediators to extend and amplify the anti-tumor effects of CAP. Using chronic myeloid leukemia K562 cells as a primary model, we demonstrate that CAP treatment induces pronounced oxidative stress, apoptosis, and sustained proliferative suppression. Importantly, exosomes isolated from CAP-treated cells exhibited enhanced anti-proliferative and pro-apoptotic activity in recipient cells compared to exosomes from untreated controls. To assess the broader applicability of this strategy, we further evaluated the effects of CAP and CAP-Exo in multiple solid tumor models, including breast, renal, and hepatocellular carcinoma cells, both in vitro and in vivo. CAP exposure consistently reduced cell viability across solid tumor cell lines, while CAP-Exo retained potent cytotoxic activity against breast cancer cells and significantly suppressed tumor growth in corresponding xenograft models without inducing systemic toxicity. Mechanistically, CAP-induced stress reprogrammed exosomal cargo, enabling the transfer of death-associated molecular signals to recipient tumor cells and thereby promoting apoptosis. Collectively, our findings indicate that CAP-modified exosomes represent a biologically active, cell-free approach that extends the anti-tumor effects of CAP treatment across both hematological malignancies and solid tumors. Rather than replacing existing therapeutic modalities, CAP-Exo may serve as a complementary strategy to enhance CAP-based cancer interventions and overcome current limitations associated with direct CAP application.

The aligned microarchitecture of extracellular matrix (ECM) has been recognized as a significant and novel hallmark of certain tumors, which is gradually uncovered to relate to their malignant progression. However, most artificial scaffolds are isotropic and fail to mimic such aligned organization of tumor-associated ECMs. To address this limitation, we developed a self-assembling peptide-based liquid crystal (LC) hydrogel model to recapitulate the aligned topology of tumor ECM, thereby establishing a platform to investigate the relationship between ECM alignment and malignant cell phenotype. The screened peptide self-assembled into aligned nanofibers via a thermal pathway, forming an LC hydrogel with engineered biological properties. When cultured within the peptide LC hydrogel, tumor cells displayed enhanced proliferation, migration, invasion, and drug-resistance, underscoring the critical role of ECM alignment in promoting aggressive phenotypes. Leveraging the LC hydrogel model, we provided initial insights into the mechanisms underlying malignant progression via Western blot, reverse transcription-quantitative PCR and RNA sequencing analyses. Moreover, by implanting LC hydrogel precultured cancer cells into C57BL/6 mice, we established a tumor model exhibiting accelerated growth. Our findings demonstrate that the self-assembled peptide LC hydrogel can recapitulate tumor ECM alignment and enable the development of rapidly progressing tumor models for cancer research and drug screening.

Thymine DNA glycosylase (TDG) is a multifaceted protein involved in base-excision repair, DNA demethylation and transcriptional regulation, with key roles in embryonic development and tumorigenesis. However, the mechanisms underlying its role in cancer progression and the therapeutic applications targeting TDG remain largely unknown. Here we demonstrate that targeting TDG induces synthetic lethality in p53-deficient cancers. We developed C-271, a first-in-class, small-molecule inhibitor that covalently binds to TDG, disrupting its DNA-binding capability. C-271 exhibits potent therapeutic efficacy in suppressing p53-deficient tumors. Mechanistically, TDG and p53 redundantly promote the transcription of DHX9, an RNA helicase that resolves double-stranded RNA (dsRNA). TDG inhibition in p53-deficient cancer cells leads to DHX9 downregulation and, thus, aberrant dsRNA accumulation, which activates the RIG-I/MDA5-MAVS sensing pathway, resulting in tumor suppression and enhanced antitumor immunity. These findings highlight the synthetic lethality between TDG and p53, positioning TDG inhibition as a promising therapeutic strategy for p53-deficient cancers.

Photodynamic therapy (PDT) that induces pyroptosis at the cell membrane has emerged as a promising paradigm for cancer immunotherapy. However, the rapid and precise induction of pyroptosis remains a significant challenge. To address this, we developed a near-infrared orthogonal excitation lanthanide theranostic nanoplatform based on lanthanide doped nanoparticles conjugated with curcumin (CUR) and cyclo(RGD-DPhe-K) peptide (LnNP@CUR-RGD). This platform enables deep-tissue, NIR-II-L imaging-guided PDT that concurrently initiates both pyroptosis and apoptosis. Upon 808 nm excitation, fluorescence at 1530 nm from LnNP@CUR-RGD allows for real-time in vivo tracking and monitoring of its localization to the cell membrane or lysosome. After subsequently switching to 940 nm excitation, the produced 362 nm emission activates CUR to generate singlet oxygen (1O2). This process initiates a dual-death mechanism: cell membrane-involved pyroptosis and lysosome-involved apoptosis, which synergistically potentiate the anti-tumor immune response. Notably, this nanoplatform achieves efficient antitumor therapy within 15 min of systemic administration, a significant acceleration compared to the 6 h required for conventional apoptosis-based PDT. This work demonstrates the considerable potential of NIR-light-triggered, targeted theranostic platforms for precise imaging-guided cancer therapy.

Paclitaxel, a clinically potent anticancer drug derived from Taxus species, faces persistent challenges in sustainable supply. Synthetic biology presents substantial opportunities for its de novo production, particularly with recent breakthroughs in elucidating its intricate biosynthetic pathways. However, its heterologous biosynthesis is significantly constrained by key bottlenecks, including pathway complexity, poor P450 expression, and inefficient metabolic flux. In this study, we explore how synthetic biology facilitates pathway decoding and reconstruction and propose strategies involving nonclassical chassis such as plant-associated cyanobacteria and filamentous fungi to enhance P450 compatibility. We also present a pragmatic framework for the rational application of state-of-the-art tools, including cell-free systems, synthetic microbial consortia, hybrid chemoenzymatic synthesis, and machine learning, to sustainably produce paclitaxel and other natural products.

Tetrahedral framework nucleic acids (tFNAs), a crucial component of DNA nanotechnology, have emerged as versatile tools in biomedical research, largely owing to their distinctive structural features and multifaceted capabilities. Recent studies have highlighted the ability of tFNAs to specifically target biological pathways, facilitate cellular uptake, and enhance therapeutic efficacy. tFNAs also show considerable potential in addressing challenges such as drug resistance and poor bioavailability. However, several challenges, including limited in vivo stability, low drug-loading capacity, and potential long-term toxicity, need to be addressed for successful clinical translation. This review comprehensively explores the latest advancements in tFNAs, focusing particularly on their applications and mechanisms in treating a wide array of diseases. From bone diseases to ophthalmic disorders, tFNAs not only promote bone regeneration but also enhance therapeutic efficacy through targeted drug delivery. They also play a role in combating infectious diseases, skin and soft tissue repair, neurological and mental disorders, hepatorenal diseases, and cancer. In addition, tFNAs have demonstrated significant potential in regenerative medicine, immunomodulation, and gene delivery. This work highlights the future potential of DNA nanotechnology in precision medicine, paving the way for next-generation therapeutic strategies.

Histology images are a cornerstone of pathology, which allow automated analysis for disease diagnosis. However, variations in staining and image acquisition processes significantly affect the performance of these algorithms. Histology image normalization is method to achieve uniformity in image color distributions, which will enhance the accuracy and consistency of automated analysis. This review was conducted with the aim of assessing normalization methods and comparing them in an empirical manner to help researchers choose the most appropriate method for their study. It also aims to assist academics and professionals involved in automated image analysis and digital pathology. This review categorizes normalization techniques into four groups: deep learning-based approaches (e.g., GANs, autoencoders, diffusion models), traditional methods (e.g., deconvolution, histogram matching), hybrid models, and a novel signal processing-based method. It also introduces a new deep learning framework for evaluating normalization strategies and experimentally compares eight state-of-the-art methods on histopathology images. The results highlight the strengths and limitations of each approach, helping researchers and professionals choose suitable methods for their needs. In addition, the review emphasizes the impact of color variation on the accuracy of computer-aided diagnosis (CAD) systems and the importance of preserving biological information during normalization. Finally, it outlines directions for future research, including integrating normalization with data augmentation and exploring information preservation beyond cancer subtype.

Hydrogen sulfide (H2S), a key endogenous gaseous mediator derived from sulfur-containing amino acid metabolism, exhibits a concentration-dependent duality in cancer. At physiological concentrations, H2S exerts antioxidant effects by activating the NRF2/Keap1 pathway and suppressing lipid peroxidation, thereby promoting tumor cell survival. In contrast, supraphysiological levels of H2S induce programmed cell death (PCD) by impairing mitochondrial homeostasis, triggering reactive oxygen species (ROS) bursts, and modifying critical proteins via persulfidation. The complex interplay between H2S and PCD pathways highlights its potential as a therapeutic target, with emerging strategies focusing on modulating endogenous H2S production and developing targeted H2S-releasing compounds. Current pharmacological approaches include inhibiting H2S-synthesizing enzymes and exogenous administration of H2S donors, which have shown promise in preclinical models and clinical trials for overcoming therapy resistance and enhancing treatment efficacy. This review establishes the integrative framework bridging H2S biology with six distinct PCD modalities, focusing on its potential therapeutic applications in cancer therapy. It further investigates the core challenges in clinical translation of H2S-based therapies, particularly the dual hurdles of achieving targeted delivery and managing concentration-dependent effects. To overcome these challenges, we outline emerging translational strategies that leverage enzyme-targeted inhibitors, repurpose H2S-modulating drugs already approved by the FDA, and integrate novel nano-theranostic platforms capable of stimulus-triggered and spatially precise release of H2S. Future research should prioritize developing intelligent delivery systems with precise spatiotemporal control, and deciphering the dynamic regulation of H2S-mediated PCD, which will be essential for advancing these mechanistic insights into precision oncology therapeutics.

Mutations in the ATRX chromatin remodeler confer a predisposition to a developmental genetic disorder and cancer, but how ATRX safeguards genome and telomere stability remains unresolved. Here, we uncover critical dependencies for the CTC1-STN1-TEN1 (CST) complex and RAD9A-HUS1-RAD1 (9-1-1) clamp in ATRX-deficient cells. ATRX-CST synthetic lethality manifests following accumulation of telomeric G-rich single-stranded DNA (ssDNA), which results in telomere loss and cell death. Conversely, we attribute ATRX-9-1-1 synthetic lethality to genome-wide ssDNA lesions, which compromise DNA replication. We further show that ATRX suppresses DNA damage during replication stress by counteracting the activity of the FAM111A protease. We demonstrate that roles of ATRX in telomere maintenance and replication are genetically separable, requiring its ATPase activity and PIP-box, respectively. We also show that such roles protecting genome stability are largely independent of the ATRX-DAXX interaction. Collectively, our data show that functions of ATRX in suppressing toxic ssDNA lesions are context-dependent and are key to global DNA replication and telomere integrity.

The developmental history of a cell fundamentally defines its identity and function. Recent advances in cell lineage tracing now enable high-resolution reconstruction of cellular ancestries in vivo, illuminating how lineage dictates fate in health and disease. This Review highlights recently developed tools, ranging from refined recombinase systems to advanced synthetic and natural barcoding approaches, that facilitate the investigation of pathological lineage programs in cancer, cardiovascular disease and aging. Moving forward, integrating permanent lineage records with single-cell multi-omics promises to provide a unified framework to decode how a cell's past shapes its present state and future potential, heralding a new era for precision medicine.

Targeting replication-associated DNA repair mechanisms, including the control of ADP-ribosylation by PARP1/2 and PARG, is a powerful therapeutic approach for cancer. However, the mechanisms by which PARG inhibition impacts DNA replication remain unclear. Here, we combine isolation of proteins on nascent DNA (iPOND) with quantitative proteomics and functional assays to investigate replication fork dynamics upon acute PARG inhibition. We find that FET family proteins (FUS, EWS, and TAF15) are recruited to replication forks in a PAR-dependent manner, forming condensates that slow fork progression and promote fork reversal. FET proteins control fork dynamics in response to some, but not all, replication stresses. FUS inactivation leads to unrestrained fork progression via RECQ1 and PRIMPOL, increased single-stranded DNA gaps, genome instability, and synthetic lethality with BRCA1 deficiency. These findings reveal that FET protein assemblies modulate replication stress responses, influencing genome stability and the cellular response to cancer therapeutics targeting PARylation pathways.

Copper, an evolutionarily conserved redox-active trace element, serves as an irreplaceable core pillar of eukaryotic life-orchestrating cuproenzyme catalysis, metabolic signaling networks, and organelle homeostasis across molecule-to-organism scales. Its strict homeostasis is a prerequisite for physiological function, and its dysregulation is a hallmark of diverse pathologies. The discovery of cuproptosis-a distinct mitochondria-centric programmed cell death-has fundamentally upended traditional paradigms of metal-mediated cytotoxicity. Cuprology, serves as an integrative interdisciplinary framework to unify copper's spatiotemporal regulation across biological hierarchies. This inherent duality governs health and disease: physiological copper sustains developmental metabolism and redox balance, while deficiency or overload drives pathogenesis spanning metabolic disorders, neurodegeneration, and cancer. Integrating foundational mechanistic insights with cutting-edge translational breakthroughs-from copper ionophores to nano-carrier-mediated precision delivery. This review synthesizes the intricate crosstalk between copper homeostasis, cuproptosis circuitry, and cellular metabolism under the Cuprology framework. It further charts a clear roadmap for targeting copper-cuproptosis axes, unlocking transformative therapeutic strategies for recalcitrant diseases and steering a new era of research into cell death and metabolic regulation.

Image-guided thermal ablation has been included in the National Comprehensive Cancer Network (NCCN) guidelines of multiple solid tumors. However, insufficient ablation of larger lesions and thermal injury adjacent to major organs and tissues limit its clinical application. Besides, sublethal hyperthermia at the margin of ablation can induce an immunosuppressive tumor microenvironment and increases recurrence risk. These physical and biological limitations are linked to each other. We developed an injectable hydrogel MR@CaP@HA with in situ thermal insulation and dual-responsive (pH/GSH) chemo-immunomodulatory delivery. MR@CaP@HA hydrogel can create a thermal insulation area with a thickness of about 5-10 mm after injection in situ, keeping the surrounding tissues under 45 °C during ablation. The disulfide-cross-linked hyaluronic acid (HA) network degrades in a glutathione (GSH)-dependent manner, inducing gel-liquid transition and controlled nanoparticle release. The released MR@CaP (calcium phosphate co-loaded with MIT and R848) nanoparticles disassemble in an acidic environment, delivering mitoxantrone (MIT) and resiquimod (R848) in a dual-responsive manner. This dual-responsive delivery system induces robust immunogenic cell death and dendritic cell maturation and achieves macrophage M1 rate of 95% in vitro and 35% in vivo. With coordinated thermal modulation and programmable drug release, the integrated therapy sterilizes residual tumor cells and achieves complete tumor eradication in 50% of animals. This work establishes a material-driven platform that overcomes thermal safety and immune resistance barriers, offering a translational strategy to enhance procedural safety and long-term efficacy.

Intercellular mitochondria transfer has emerged as a new form of cell-to-cell communication with profound consequences for cellular fate. A growing body of evidence defines mitochondria transfer between cells as a new pathological program in which cancer cells appropriate functional mitochondria from donor cells, thereby co-opting conserved physiological mechanisms of energy allocation to gain bioenergetic and phenotypic advantages. Our recent work demonstrates the prevalence of mitochondria transfer at the nerve-cancer interface, with neurons, though not exclusively, serving as a prominent source of the organelle. This suggests an unrecognized role of the nervous system in systemic energy redistribution and indicates that tumors may exploit this ancient, physiologically grounded mechanism to fuel progression and metastasis.

Why are clonal neoantigens paradoxically rare in established tumors? Why does tumor-associated antigen (TAA) burden inversely correlate with T-cell infiltration? Why do damage-associated molecular pattern (DAMP)/danger signal-inducing interventions (messenger RNA (mRNA) vaccines, stereotactic body radiotherapy (SBRT), immunogenic chemotherapy) enhance checkpoint blockade activity in "cold" tumors? We propose the "DEA Hypothesis" to resolve these paradoxes: durable antitumor immunity for established tumors requires three convergent signals-(1) Disinhibition of T cells through checkpoint blockade, (2) Enhanced recognition via the presentation of clonal neoantigens, and crucially, (3) Alerting the host through release of DAMPs/danger signal(s). We frame cancer as a subversion of an ancient "Guardian" system-the emergency hematopoietic response that evolved to orchestrate immune defense following epithelial breach or major trauma. Tumors hijack this system by promoting restorative hematopoiesis at the expense of immunity and masquerading as chronic wounds. This framework explains key observations: the paucity of clonal neoantigens reflects successful immunoediting over years of tumor evolution, not failed presentation. TAAs persist in cold tumors because insufficient DAMPs limit the "Decision to Attack"; and mRNA vaccines, recently SBRT, and immunogenic chemotherapy succeed in cold tumors by providing the missing Alert. For cold tumors, rational immunotherapy must extend beyond checkpoint blockade to reconstitute the Alert signal-not merely releasing the brakes but providing the ignition to reawaken the ancient Guardian.Graphic abstract: The DEA hypothesis proposes that effective cancer immunotherapy requires three convergent signals: Disinhibition (checkpoint blockade), Enhanced recognition (clonal neoantigens), and Alerting the host (DAMP release). Panel I depicts immune decision points in tumor-draining lymph nodes. Panel II shows mutation-driven DAMP generation via the Poly(ADP-Ribose) Polymerase 1 - High Mobility Group Box 1 (PARP1-HMGB1) axis. Panel III illustrates the three-signal model converging on type 1 conventional dendritic cells activation. Panel IV demonstrates how immunoediting shapes stage-specific therapeutic strategies for cold, warm, and hot tumors.

While conventionally viewed as a defense against external pathogens, we posit that the adaptive immune system likely evolved primarily to preserve internal epithelial order. We propose that adaptive immunity, with combinatorial diversity emerging ~0.5 billion years ago during the Cambrian explosion, was based on extant and established complex cellular interactions, derived over the previous 3.5 billion years of life's history. These coordinate cellular processes originated to police epithelia against aberrant or rogue cells-inherent to complex multicellularity. Primordial surveillance relied on maintaining epithelial integrity and specialized immune sentinels. Early deposition of tissue-resident γδ T and natural killer cells detects abnormalities and emergent stress signals as damage-associated molecular pattern molecules (DAMPs), independently of classical major histocompatibility (MHC) presentation to αβ T cells. Trained immunity in tissue-resident macrophages, stroma, and epithelia reciprocally respond and are tailored by nascent and highly selected tissue-resident memory cells.

Epi-immunotherapy holds substantial potential in overcoming intrinsic and acquired resistance in tumor immunotherapy. However, most epi-immunotherapy strategies fail to match the dynamics of immune responses due to a lack of temporal regulation, resulting in short-lived efficacy. In addition, the instability and inefficient intracellular delivery of gene regulatory drugs, especially siRNA, further limit the therapeutic efficacy of epi-immunotherapy. Here, we propose and implement a temporal immunomodulation via methylation epigenetics (TIME) strategy through the combined intravenous administration of the FDA-approved small-molecule drug azacitidine and a BP/siPRMT1 nanocomplex. Azacitidine serves as the "Ignite" module, while BP/siPRMT1 was prepared by simple mixing of chemically synthesized boronated polypeptide (BP) with siPRMT1 at room temperature and functions as the "Sustain" module. By rapidly reducing intratumoral methylation levels through azacitidine, the expression of major histocompatibility complex class I (MHC-I) and secretion of interferon gamma (IFN-γ) were significantly enhanced, thereby igniting a hot tumor microenvironment. Subsequently, BP/siPRMT1 maintained high levels of MHC-I expression and IFN-γ secretion, sustaining the azacitidine-ignited antitumor immunity. Notably, the TIME strategy increased the proportion of MHC-I-positive tumor cells by 6.7-fold in tumor tissues. This "Ignite-Sustain" feature enables rapid immune sensitization and long-term immune maintenance. Compared to the delivery of an epi-drug or nucleic acid drug alone, the TIME strategy effectively expands the critical time window for immune activation, achieving durable and controllable tumor immunotherapy.

Facile and quantitative detection of liquid biopsy biomarkers such as microRNAs offers significant potential for precision healthcare; however, conventional biosensing methods rely on enzyme- or label-based workflows that are costly, time-consuming, and labor intensive. Microwave biosensors, particularly split-ring resonators (SRRs), offer an attractive alternative as they enable label-free, noncontact electromagnetic detection through permittivity measurements and are compatible with printed-circuit-board manufacturing. However, the sensitivity of conventional SRR platforms remains insufficient for clinically relevant biomarker detection. Here, we introduce an enzyme-free, label-free microwave biosensing architecture that integrates SRRs with microfluidic channels containing localized bioreceptor-functionalized hydrogel micropillars. Target hybridization within the hydrogel micropillars induces localized changes in complex permittivity, which are transduced into concentration-dependent shifts in the resonant frequency of the SRR capacitive gap. As a proof of concept, the platform is applied to detect the cancer-associated biomarker miR-16-5p using peptide nucleic acid (PNA) probes, which were selected for their neutral backbone, enzymatic stability, and strong hybridization affinity. The hydrogel micropillars act as three-dimensional scaffolds that enhance probe loading and maximize volumetric electromagnetic interaction, representing a departure from conventional planar biointerfaces. Compared with equivalent planar systems, this architecture achieves approximately a 20-fold improvement in detection limit, reaching subnanomolar sensitivity without any amplification or labeling while maintaining single-nucleotide specificity and strong device reproducibility. Beyond being the first demonstration of SRR-based miRNA detection, this work establishes a general strategy for three-dimensional microwave biosensing and positions hydrogel-interfaced resonators as a next-generation platform for sensitive, selective, label-free, and reusable biosensors.

The clinical application of nucleic acid aptamers is hindered by rapid degradation and poor targeting in physiological conditions. Multivalent assembly can improve their performance, yet key structural rules for stability are not fully clarified. Here, we fabricate a symmetric tetravalent aptamer-streptavidin conjugate (Tetra-XQ@SA) for cancer theranostics with greatly enhanced stability. Thymine bases, longer sequences, and double-stranded structures collectively strengthen its nuclease resistance, enabling stable storage for 96 h in DNase I and 72 h in serum. Compared with free aptamers, Tetra-XQ@SA shows 2-fold higher binding affinity to cancer cell CD71 receptors, 7-fold greater doxorubicin loading, and improved antitumor activity. It achieves prolonged tumor retention and an ∼73% tumor inhibition rate in vivo. This study reveals the structure-stability relationship of aptamer assemblies and offers a versatile platform for targeted cancer therapy.

Immune checkpoint inhibitors (ICI) have had a transformative impact on cancer treatment. However, many patients still fail to respond to ICIs, underscoring the need to identify novel drivers of resistance. In this issue, Ou and colleagues identify single-stranded DNA binding protein 4 (SSBP4) as one such candidate, demonstrating that it promotes intratumoral cholesterol biosynthesis to suppress lymphocyte recruitment and enable immune evasion. This work therefore nominates SSBP4 as a novel metabolic target to enhance antitumor immunity and expand ICI efficacy. See related article by Ou et al., p. XX .

Neutrophils are an ideal drug carrier because of their specific targeting and effective accumulation at sites of inflammation. However, the clinical application of neutrophils as drug carriers is constrained by the inability to sustain ex vivo culture and their short lifespan. Herein, we developed an in situ construction strategy to fabricate neutrophil-nanoparticle biohybrid systems (NE@NPs) in vivo by specifically hijacking the pro-inflammatory neutrophils in blood using thermoresponsive drug-loaded NPs. The pro-inflammatory neutrophils effectively transport these NPs to the tumor microenvironment across the vascular barriers, followed by the release of the NPs through neutrophil extracellular traps (NETs). Subsequent near-infrared (NIR) light-induced spatiotemporally drug-controlled release improved the therapeutic response. The therapeutic efficacy of in situ-constructed NE@NPs is successfully confirmed using multiple mouse models of cancer, pulmonary, and skin infections. This study illustrates that the in situ construction strategy of neutrophil-NP hybrid systems in vivo may be a promising approach for improving inflammation-associated disease treatment.

Traditional tumor immunology research has predominantly focused on circulating recruited immune cells. In contrast, tissue-resident immune cells-a unique subset that stably resides in normal tissues and the tumor microenvironment (TME) for extended periods without entering the systemic circulation-exhibit distinct transcriptional, epigenetic and functional profiles, and their roles in tumor initiation, progression and therapeutic response remain incompletely elucidated. Focusing on the representative subsets of resident T cells, natural killer (NK) cells and macrophages, this review defines the origins of tissue-resident immune cells and elucidates the transcriptional regulatory networks (including TGF-β/IL-33 and RUNX3) that control their differentiation, maintenance, and tissue retention. Meanwhile, we systematically compare tissue-resident immune cells with recruited immune cells and summarize the canonical cell surface markers of resident cells such as CD69 and CD49a. We further reveal the bidirectional plasticity between tissue-resident and recruited immune cells: circulating T cells and monocytes can differentiate into resident subsets under the influence of local cytokine signals such as TGF-β and IFN-γ, while a fraction of tissue-resident immune cells retain the capacity to re-enter the circulation and migrate to distant tissues under inflammatory conditions. On this basis, we dissect the molecular mechanisms underlying the dual pro-tumor and anti-tumor effects exerted by the tissue-resident immune cells in the TME, which are mediated through cytokine regulation, direct contact-mediated cytotoxicity, and indirect modulation of recruited immune cells. Finally, we summarize the multifaceted impacts of conventional cancer therapies (radiotherapy, chemotherapy and targeted therapy) on tissue-resident immune cells, propose innovative therapeutic strategies targeting tissue-resident immune cells, and outline the directions for the development of next-generation molecular targeted anticancer agents. Collectively, this comprehensive review clarifies the mechanistic basis of the dual roles of tissue-resident immune cells in tumor immunity, uncovers their functional plasticity and co-evolutionary mechanisms with the TME, and provides a theoretical framework and novel therapeutic targets for overcoming immune escape in cold tumors.

WD40-repeat (WDR) proteins constitute one of the largest and most functionally diverse scaffold families in eukaryotes. By folding tandem WD repeats into β-propeller domains, they provide modular interaction surfaces that assemble multi-protein complexes governing transcription and epigenetic control, ubiquitin-dependent proteostasis, RNA metabolism, and cell-cycle progression. Despite their pervasive involvement in oncogenic signaling and hallmark cancer phenotypes, the cancer field still lacks an integrated framework that connects WDR structural logic to context-dependent mechanisms and, critically, to actionable biomarkers and therapeutic strategies; existing evidence remains dispersed across tumor types and molecular pathways. This review synthesizes current knowledge to address that gap. We first summarize core structural principles of WD40 β-propellers and explain how multivalent binding and partner selection enable WDR proteins to function as assembly platforms in oncogenic networks. We then consolidate mechanistic evidence showing how representative WDR proteins shape malignant state transitions, including sustained proliferation, survival under stress, epigenetic plasticity, invasion and metastasis, and therapy resistance, by rewiring chromatin programs, ubiquitination circuits, and RNA and translation outputs across cancers. Finally, we highlight translational progress and opportunities. Overall, this review integrates the fragmented WDR research into a clinically oriented framework that clarifies their potential as biomarkers for early detection, stratified diagnosis, and treatment-response prediction, and delineates druggable entry points and rational combination strategies, thereby providing a translational roadmap to enable more precise cancer diagnosis and more effective targeted therapies in the future.

The inevitable splenic sequestration of nanomedicine represents a major delivery challenge that compromises tumor accumulation and may contribute to off-target burden. Rather than attempting to eliminate this fate, we propose to functionally repurpose it into an immunologically productive process. Distinct from conventional spleen-targeted strategies, we aimed to modulate the biological fate of tumor-targeted nanomedicines by engineering their surface with screening-selected hybrid cell membranes. A tumor cell membrane-to-erythrocyte membrane ratio of 10:1(TRM) was identified as a representative balance point that preserves optimal tumor-associated accumulation while biasing a fraction of off-target nanoparticles (NPs) toward the spleen. Zinc imidazolate framework-8 (ZIF-8) NPs were employed as a model therapeutic core. The resulting hybrid cell membrane-camouflaged nanoplatform (TRM@ZIF-8) exhibited immune-associated effects, where the tumor-associated fraction was linked to inflammasome-related signaling and inflammatory cell death-associated responses, while the spleen-biased fraction was associated with activation of splenic antigen-presenting cells. This behavior is consistent with the collective influence of membrane-associated protein features, including functions related to cluster of differentiation 47(CD47) and Band 3, together with the intrinsic immunostimulatory properties of the ZIF-8 core. Importantly, splenectomy markedly attenuated the therapeutic effect, providing organ-level evidence that spleen involvement is functionally associated with the observed antitumor immune response and tumor growth suppression. This work provides a proof-of-concept framework for guiding off-targeted nanomedicine repurposing in spleen for cancer immunotherapy, rather than regarding it solely as a delivery limitation.

Chronic tumor-associated inflammation can impair dendritic-cell (DC) antigen presentation and limit immune engagement, posing a barrier to ferroptosis-based therapeutic strategies that often intensify oxidative stress. Here we develop an inflammation-compatible ferroptosis-inducing therapeutic platform by co-integrating an NF-κB-modulating small molecule, celastrol, with an iron-coordinated mesoporous polydopamine carrier (mPDA-Fe-Cel). The platform combines 808-nm photothermal heating with iron redox cycling to promote lipid peroxidation, while concurrently attenuating inflammatory signaling programs, thereby supporting ferroptosis-associated tumor cell damage in a more immune-permissive context. In tumor cells, mPDA-Fe-Cel increases labile Fe²⁺, depletes glutathione, downregulates GPX4, and amplifies lipid peroxidation, consistent with ferroptosis-associated cell death and accompanied by immunogenic cell death hallmarks, including calreticulin exposure, HMGB1 release, and ATP secretion. In parallel, transcriptomic and protein-level analyses indicate attenuation of NF-κB-linked inflammatory programs, with prominent suppression of a CXCL8-centered signature, together with reduced activation of Toll-like receptor and NOD-like receptor pathways. Functionally, tumor-conditioned cues generated by the nanoplatform support DC maturation and antigen-presentation-related phenotypes (MHC-I/MHC-II and costimulatory markers), coinciding with enhanced intratumoral CD8⁺ T-cell infiltration and improved antitumor efficacy in murine models with favorable biosafety. Collectively, this work outlines a materials-based strategy that couples ferroptosis induction with inflammatory modulation to facilitate immune-compatible cancer nanotherapy.

The human sodium-dependent multivitamin transporter (SMVT, SLC5A6) mediates cellular uptake of essential metabolic cofactors, including biotin, pantothenate, and lipoate. Its dysfunction is associated with neurological disorders, metabolic abnormalities, and cancer. However, the molecular mechanism underlying its multi-substrate transport has remained elusive. Here, we present cryo-electron microscopy structures of human SMVT in three conformational states: occluded, outward-open, and inward-open. These structural snapshots capture the complete transport cycle and reveal a conserved substrate-binding pocket near a kinked transmembrane helix (TM1). Within this pocket, substrate carboxyl groups are electrostatically anchored, while distinct chemical moieties interact with specific polar and hydrophobic residues. Functional assays identify key binding residues and elucidate the pathogenic effects of disease-associated mutations. Further structural analysis delineates the principles of substrate discrimination within the SLC5 transporter family. Together, our work provides a structural framework for SMVT's polyspecificity and lays a foundation for understanding related diseases and developing targeted therapeutic strategies.

Targeted protein degraders (TPDs), including proteolysis-targeting chimeras (PROTAC) and molecular glue degraders (MGD), are among the most promising small-molecule-based drug treatments in oncology. The May 2026 U.S. Food and Drug Administration (FDA) approval of vepdegestrant provides a regulatory milestone for heterobifunctional protein degradation and for PROTAC therapeutics. First-generation TPDs were developed for oral delivery; however, the intrinsic physicochemical properties of TPDs impose constraints on their oral bioavailability, systemic exposure, target-site accumulation, and therapeutic efficacy. As the field transitions toward a second wave of TPD development, nanoparticle-based targeted protein degraders (nano-TPD) are gaining momentum for broadening the therapeutic landscape of protein degradation. In this context, drug delivery systems offer opportunities to overcome key translational barriers by improving pharmacokinetics, tissue distribution, target site localization, cellular uptake, and therapeutic index. Here, we provide an overview of TPD discovery, from early laboratory to (pre-) clinical progress, discuss translational challenges, and suggest advanced drug delivery solutions to help realize the full potential of TPD therapies.

Appendiceal adenocarcinoma (AA) is a rare cancer with limited treatment options. KRAS is the most commonly mutated gene in AA and a promising therapeutic target, but its preclinical and translational relevance in AA remains unclear. We evaluated KRASG12D-specific (MRTX1133) and pan-KRAS inhibitor (RMC-6236) in KRASmut organoid and PDX models of AA. Tumor-intrinsic and microenvironmental responses were characterized using multi-omics profiling. Clinical outcomes were also assessed in cohort of 15 patients with AA treated with KRAS inhibitors. MRTX1133 was highly effective for KRASG12D organoids (IC50 = 4.1 nM); both KRASG12D and KRASG12V organoids were sensitive to RMC-6236 (IC50 = 4.4 nM vs. 0.5 nM, respectively). In PDX models of peritoneal carcinomatosis from AA, MRTX1133 reduced tumor growth in the KRASG12D model TM00351, and RMC-6236 reduced tumor growth in KRASG12V model AAPDX-16. Pathologic evaluation showed dramatically reduced tumor cellularity, proliferation, and pERK expression as well as induction of apoptosis. Gene Set Enrichment Analysis (GSEA) demonstrated consistent downregulation of E2F targets and RAS/ERK signaling across both models, confirming on-target KRAS inhibition, alongside upregulation of EMT (Epithelial-to-Mesenchymal Transition), suggesting a shared resistance-associated transcriptional program. Interferon-alpha and interferon-gamma signaling were significantly upregulated following pan-KRAS inhibition, implicating TME (Tumor Microenvironment) immunomodulation as a downstream consequence of KRAS inhibition in AA. scRNA-seq analysis of the TME showed a dramatic shift from normal to inflammatory cancer-associated fibroblasts (CAFs) following KRAS inhibition. Upregulation of interferon alpha and gamma pathways was also observed, suggesting that KRAS inhibition can activate innate immune response in the setting of peritoneal metastases. Among 15 patients treated with KRAS inhibitors (8 G12D, 5 G12C, 2 pan-KRAS), all evaluable pateints had a biochemical response by serum markers and clinical benefit by imaging criteria (1 CR, 1 PR, 12 SD). While effective suppression of RAS/ERK signaling by KRAS inhibitors reduces tumor growth, adaptive activation of EMT pathway may mediate resistance in KRASmut AA. Additionally, KRAS inhibition remodels TME and may enhance innate immune signaling. These findings support the continued clinical development of KRAS inhibitors in AA and provide a rationale for combination strategies targeting resistance pathways and stromal remodeling.

We present the 18 F-FDG PET/CT findings of pancreatic metastasis in a 60-year-old man with a history of rectal cancer, who presented with abdominal discomfort and elevated carcinoembryonic antigen (CEA). 18 F-FDG PET/CT revealed a hypermetabolic pancreatic nodule with ductal dilation. Histopathology confirmed metastatic intestinal-origin adenocarcinoma. This case highlights the diagnostic challenge in differentiating metastasis from primary pancreatic cancer, particularly given the uncommon presentation of solitary pancreatic metastasis and its atypical imaging features. A comprehensive evaluation is essential for accurate diagnosis.

This paper introduces a continuous transient photoelectrochemical (PEC) biosensing strategy for rapid anti-vascular endothelial growth factor (VEGF) immobilization and quantification of VEGF in serum for colorectal cancer (CRC) diagnosis. A gold electrode modified with cis-bis(isothiocyanato) (2,2'-bipyridyl-4,4'-dicarboxylato) (4,4'-di-nonyl-2'-bipyridyl)ruthenium (II) (Z907) (Au/Z907) was used. By optimizing the electrolyte composition and applied potential under continuous high-frequency light switching, stable transient photocurrent bands were produced, enabling real-time sensing with intrinsic signal amplification and improved precision. Under optimized conditions (1 mM phosphate-buffered saline, 0.15 V bias, 25 W white-light illumination, and 2 Hz modulation for 4 min), anti-VEGF antibodies were rapidly immobilized on the Au/Z907 electrode. After blocking nonspecific binding sites with bovine serum albumin, the biosensor was used to quantify serum VEGF in clinical samples obtained in Taiwan from healthy donors (n = 11) and patients with tubular adenoma (n = 12), serrated adenoma (n = 11), untreated CRC (n = 11), and treated CRC (n = 3). Comparison with a commercial enzyme-linked immunosorbent assay kit revealed no significant differences among the five groups (p > 0.05, Student's t-test), validating the accuracy of the proposed platform. Receiver operating characteristic analysis further demonstrated excellent diagnostic performance. Overall, this work presents a cost-effective and instrumentation-light PEC biosensing platform capable of rapid antibody immobilization, calibration, and serum analysis on a single electrode, with potential applications in point-of-care testing in community hospitals and resource-limited settings.

A 44-year-old man with a history of ulcerative colitis and primary sclerosing cholangitis presented with fecaluria and was found to have rectal and cecal masses, later histologically confirmed as mucinous adenocarcinomas forming rectovesical fistulas. Despite chemotherapy reducing the primary tumor size, elevated serum CEA levels prompted further imaging. An 18 F-FDG PET/CT scan showed no new lesions, but a subsequent 99m Tc-FAPI-46 scintigraphy revealed diffuse peritoneal involvement. During surgery for bowel obstruction, multiple metastatic deposits were identified. This case underscores the superior detection capability of 99m Tc-FAPI-46 SPECT/CT in assessing peritoneal carcinomatosis in colorectal mucinous adenocarcinomas compared with 18 F-FDG PET/CT.

Situs inversus totalis (SIT) is a rare congenital anomaly characterized by the mirror-image transposition of the normal anatomical positions of major visceral organs and vascular structures. The reversed placement of cardiovascular structures observed in SIT may lead to additional technical challenges in endovascular treatment procedures. This paper presents the transarterial radioembolization (TARE) treatment applied to a 59-year-old man with SIT diagnosed with liver metastasis from rectal cancer. A literature review revealed no publications regarding the application of TARE in patients with SIT. This case is significant in shedding light on interventional oncologic treatments in patients with similar anatomical variations.

Cell competition is an emerging mechanism in which mammalian tissues maintain homeostasis by eliminating less fit (loser) cells through direct interactions with fitter (winner) neighbouring cells. In cancer, these competitive interactions may drive tumour evolution; however, spatial organisation and clinical relevance of these events remain poorly understood. One mechanism by which winner cells eliminate loser cells is engulfment, resulting in cell-in-cell (CIC) formation. Although CICs have been observed in many tumour types for over a century, their cellular composition, spatial context, interactions with the tumour microenvironment, and biological significance in human cancers remain unclear. Here, we systematically characterised the cellular identity and functional states of CICs in situ, examined their spatial interactions within the tumour microenvironment, and assessed their clinical relevance using spatially resolved single-cell data from a large cohort of colorectal cancer patients. We demonstrated that CICs occurred predominantly between cancer cells but also involved cancer stem cell (CSC)-like populations and cytotoxic T cells. Engulfed (inner) cancer and CSC-like cells displayed molecular features consistent with a loser-cell phenotype, including increased apoptosis and reduced proliferation, whereas outer cancer cells exhibited winner-cell features such as upregulated glycolysis. Live-cell time-lapse experiments demonstrated that glucose accumulated in inner cells during lysosomal degradation following cell engulfment. Spatial analysis further revealed distinct CIC neighbourhoods, which we defined based on proximity to engulfment events. Cells within these regions, particularly CSC-like cells and cytotoxic T cells, exhibited increased metabolic stress, suggesting local competition for nutrients. Importantly, the presence of cytotoxic T cells within CIC neighbourhoods and spatial co-occurrence between cancer cells and CSC-like populations were associated with improved patient outcomes. Together, our findings demonstrate that cell engulfment defines spatially organised competitive niches and may reflect cell competition within complex tumour microenvironments.

Previous studies have suggested a possible association between Helicobacter pylori (H. pylori) infection and an increased risk of colorectal cancer (CRC). We examined the associations of H. pylori infection and H. pylori treatment with incident CRC risk and evaluated whether these associations were modified by genetic susceptibility. This study was based on two randomised trial cohorts: the Shandong Intervention Trial (SIT; n=3365; 1995-2024) and the Mass Intervention Trial in Linqu, Shandong Province (MITS; n=180 284; 2011-2024). Within MITS, we further conducted a case-cohort study to assess CRC risk according to seropositivity for H. pylori virulence factors and host genetic predisposition. Compared with H. pylori-negative individuals, H. pylori-positive individuals who did not receive antibiotic treatment had a significantly higher risk of CRC (SIT: HR=2.96, 95% CI 1.30 to 6.71; MITS: HR=1.27, 95% CI 1.04 to 1.55). The increased risk was particularly evident among individuals seropositive for four H. pylori-specific antigens (CagA, HpaA, Omp and HP0305) and those at high genetic risk (top decile of the Polygenic Risk Score). In SIT, H. pylori treatment was associated with a significantly reduced CRC risk over 29.4 years of follow-up (HR=0.47, 95% CI 0.22 to 0.99), with a greater reduction observed among participants with successful eradication (HR=0.38, 95% CI 0.15 to 0.94). In MITS, no overall benefit was observed 13.8 years after treatment (HR=1.17, 95% CI 0.95 to 1.43). However, a protective effect was apparent among individuals at high genetic risk and among those seropositive for key H. pylori-specific antigens. In post hoc observational analyses of two established trial cohorts, H. pylori infection was associated with an increased risk of CRC. H. pylori treatment reduced CRC risk in SIT, whereas in MITS, the benefit appeared to be limited to individuals at high genetic risk or those infected with specific virulence factor subtypes.

Elevated intratumoral immune inflammation prior to treatment is typically associated with better outcomes in hot tumors treated with immune checkpoint inhibitors (ICIs). However, we observed a paradox in pMMR/MSS locally advanced rectal cancer (LARC) patients, where a subset with elevated baseline immune inflammation exhibited worse outcomes after combined radiotherapy and ICI treatment compared with patients with minimal immune inflammation. To investigate this counterintuitive finding, we performed paired scRNA-seq and scTCR-seq on longitudinally collected samples, including tumor biopsies (pre-treatment, post-radiotherapy, and post-immunotherapy) and peripheral blood mononuclear cells (pre-treatment and post-immunotherapy), from 20 pMMR/MSS LARC patients treated with sequential radiotherapy and ICI therapy (NCT06493240). We propose the concept of clonal entrapment to explain this phenomenon. Specifically, our profiling results reveal that increased HLA-DQA2 expression in dendritic cells and upregulated GDF15 expression in treatment-resistant tumor cells correlate with the restricted expansion of novel tumor-reactive TCR clonotypes. Consequently, the immune response is limited primarily by pre-existing TCR clonotypes within the tumor, especially those partially expanded under chronic inflammation, leading to the expansion of TCR clonotypes derived mainly from pre-treatment CD8+ T cell pools following ICI therapy. By identifying this feature of the pMMR/MSS LARC microenvironment, our study provides a high-resolution framework for understanding resistance to sequential radiotherapy and ICI therapy.

The gut microbiome shapes cancer progression and treatment responses, yet scalable microbiome-targeted interventions remain limited. We screened commercial probiotics for activation of the host aryl hydrocarbon receptor (AhR) and identified the yeast Saccharomyces boulardii as a consistent AhR activator. In an immunocompetent syngeneic colorectal cancer model, daily oral gavage of S. boulardii slowed growth of established subcutaneous tumors without detectable tumor colonization. Integrated profiling of the gut microbiome, circulating metabolites, cytokines, and tumor transcriptomes revealed a coordinated systemic response. S. boulardii increased microbial diversity and functionally rebalanced the gut microbiota, enriching taxa with lower genome-encoded biosynthetic autonomy. These changes were accompanied by elevated plasma levels of several indole metabolites, including the AhR agonists 5-hydroxyindole-3-acetic acid (5-HIAA) and indole-3-propionic acid (IPA). Targeted LC-MS/MS showed that S. boulardii can produce 5-HIAA under culture conditions, whereas IPA was not detected, suggesting that increased plasma levels of these metabolites may arise through a combination of probiotic activity and broader microbiome-associated processes. Circulating IL-17A and CTLA-4 were reduced, and tumors exhibited downregulation of programs linked to invasion, inflammation, and KRAS signaling. Multi-omics integration showed strong covariation across microbial, metabolic, immune signaling, and tumor compartments, highlighting coordinated cross-compartment responses during S. boulardii-associated tumor suppression.

Fusobacterium nucleatum has emerged as a pathobiont that associates oral dysbiosis with systemic diseases through coaggregation, hematogenous dissemination, and immune modulation. This review provides molecular insights through which they are involved in systemic diseases such as colorectal cancer, adverse pregnancy outcomes, cardiovascular diseases, neurodegenerative disorders, and diabetes mellitus. Key virulence factors include the adhesins of FadA, Fap2, and RadD, lipopolysaccharides, and outer membrane vesicles, which mediate epithelial invasion and endothelial permeafbility and facilitate immune suppression through TLR4-NF-κB, β-catenin/Wnt, and MAPK signaling pathways. These interactions result in impaired tissue homeostasis, propagate chronic inflammation, and promote oncogenic and metabolic modulation. Systemic pleiotropy of F. nucleatum is further substantiated by its involvement in chemoresistance, placental dysfunction, vascular inflammation, and neuronal injury, substantiating its systemic pleiotropy. Emerging therapeutic strategies, such as blocking adhesins, neutralizing outer membrane vesicles, microbiome manipulation, and using CRISPR-based clearance, provide precision techniques for mitigating diseases. Therefore, this review identifies F. nucleatum as the primary microbial mediator of oral-systemic pathology and its translational significance in the development of targeted antimicrobial and host-directed therapies.

Growing evidence suggests a pivotal role of the microbiome in tumorigenesis, extending beyond genetics. Apc(Min/+) mice exhibit reduced tumor load when housed in germ-free conditions. Nevertheless, how genetic factors shape microbiota and how dysbiosis fits into the genetic paradigm of intestinal carcinogenesis remain elusive. Epithelial stemness is regulated by Wnt/Apc/β-catenin pathway, whereas Apc mutations and Hippo signaling are associated with tumor growth. Invasive pathobionts emerge from microbiota as a result of epithelial barrier dysfunction. We hypothesize that the emergence of invasive pathobionts and dysbiosis of epithelial microbiota contribute to increased cancer stemness. The epithelial and fecal microbiota are longitudinally monitored in Apc(Min/+) and wild-type littermates born to wild-type surrogate dams. Segregation of epithelial microbiota between Apc(Min/+) and wild-type mice was observed as early as eight weeks after birth, whereas fecal microbiota diverged at 20 weeks of age. Epithelial dysbiosis and barrier defects were observed in Apc(Min/+) mice, characterized by intraepithelial Escherichia coli with invasive features. While antibiotic treatment reduced cancer burden, invasive E. coli infection promoted tumorsphere formation. Higher expression of Vgll3 and Tead4 (Hippo effectors) and Cd44 (a cancer stemness marker) was observed in bacteria-infected tumorspheres. Mechanistically, bacteria augmented epithelial clonogenicity by enhancing VGLL3/TEAD4-mediated CD44 promoter activity. Invasive E. coli genetic signatures were verified in 86% of human colorectal carcinoma specimens, and a positive correlation with TEAD4 expression was observed. In conclusion, Apc mutation drives the expansion of invasive pathobionts to promote cancer stemness via a VGLL3/TEAD4/CD44 axis. Bacteria-targeting interventions could be an alternative strategy for patients with hereditary tumors.

Chemoresistance to 5-fluorouracil (5-FU) remains a critical barrier in colorectal cancer (CRC) management. This study integrates multi-omics data from 26,192 human samples to elucidate the RNA methylation-mediated crosstalk between tumor-associated macrophages (TAMs) and tumor cells. Machine learning models were able to effectively stratify patients by risk and identified a core signature of six genes (including SCG2), whose expression patterns were associated with poor prognosis and chemotherapy resistance-related phenotypes. Mechanistically, 5-FU elevates m6A modification in TAMs, polarizing them toward the M2 phenotype. SCG2 mRNA methylation promotes TNF-α ubiquitination, reducing its levels and thereby sustaining NF-κB activation in tumor cells to drive PCD resistance. The core candidate genes (CCGs) model effectively predicts survival outcomes. Targeting SCG2 represents a novel strategy to reverse chemoresistance by disrupting TAMs-tumor crosstalk, offering actionable targets for personalized therapy optimization.

Tumors reprogram lipid metabolism in distant tissues to support their growth. In adult Drosophila, gut tumors secrete the PDGF/VEGF-like factor Pvf1, which activates the TORC1-Hnf4 pathway in hepatocyte-like oenocytes. This drives production of very long-chain fatty acids and wax esters essential for tracheal growth around the tumor. Blocking Hnf4 or the elongase mElo in oenocytes strongly suppresses tracheogenesis, tumor progression, and cachexia-like organ wasting, while extending host lifespan. The same pathway also controls tracheal development in healthy flies. Lipoprotein receptor LpR2 depletion in oenocytes rescues the observed tumor induced tracheal tracheal remodeling. This tumor-host interaction is conserved: VEGF-A induces lipid metabolism genes in human hepatocytes, and lung tumor-bearing mice show elevated hepatic Hnf4 and Elovl7. Altogether, this study reveals a non-autonomous role of the TORC1-Hnf4 axis in lipid-mediated tumor progression and identifies potential therapeutic targets for cancer-associated metabolic dysfunction.

The β-catenin destruction complex (BDC) regulates WNT-β-catenin signaling and is a prime therapeutic target in colorectal cancer, yet its biochemical complexity has hindered mechanistic understanding. We mapped the sequence-function landscape of the BDC using tiled base editor screens across its components CTNNB1, AXIN1, APC and GSK3B. Amongst ~150 previously unreported mutations that affected WNT signaling, we discovered gain-of-function and separation-of-function alleles that reveal mechanisms of complex assembly, including a β-catenin region regulating TCF/LEF transcription factor binding. Critically, we found that the AXIN1-β-catenin interface controls signaling flux through the oncogenic BDC found in APC-mutant cancers. In cells expressing truncated APC, β-catenin itself scaffolds BDC assembly, establishing a substrate-assisted autoregulatory mechanism. This architecture represents an unexploited therapeutic vulnerability: strengthening the AXIN1-β-catenin interaction restores destruction complex function and impairs the growth of colorectal cancer cells. Our mutational resource provides a foundation for mechanistic understanding and therapeutic targeting of the WNT pathway.

Colorectal cancer with liver metastases remains a clinical challenge. CXCL13 is widely recognized as a biomarker of immunotherapy response. However, the functional heterogeneity (protumor vs. antitumor) of CXCL13+CD4+ T-cell subsets has long been controversial. Through integrated analysis of single-cell RNA sequencing data from colorectal cancer clinical samples and pan-cancer datasets, combined with experimental validations, we first identified a prometastatic neuromedin B+ (NMB+)CXCL13+CD4+ T-cell subset and uncovered a mechanism by which this subset regulates tumor cell "senescence-malignant transition," the NMB-NPSR1 axis. These NMB+CXCL13+CD4+ T cells induced senescence in NPSR1+ malignant cells via NMB secretion, leading to enhanced invasiveness and migration despite reduced proliferation. Activation of NPSR1 triggered the Wnt signaling pathway and epithelial-mesenchymal transition, thereby enhancing cellular malignancy. This NPSR1+ senescent subpopulation also recruited endothelial cells and disrupted tight junction integrity, fostering a prometastatic microenvironment. As a proof-of-principle study, the combination of the NPSR1 inhibitor SHA68 and anti-PD-1 demonstrated remarkable antitumor effects using mouse models of colorectal cancer metastasis. Overall, this study uncovered the role of NMB+CXCL13+CD4+ T cells in promoting tumor cell senescence while influencing tumor metastasis, offering potential clinical implications for the diagnosis and treatment of metastatic colorectal cancer.

Accumulating evidence indicates that peritoneal macrophages, comprising resident peritoneal macrophages (resMØ) and monocyte-derived nonresident macrophages (moMØ), contribute to peritoneal tumor progression by promoting tumor cell proliferation and migration and driving immunosuppression. However, the mechanisms governing the expansion of resMØs and moMØs, as well as their differential contributions to the peritoneal macrophage pool in tumor-bearing mice and to tumor growth, remain to be elucidated. Using a mouse model of colorectal cancer peritoneal metastasis, induced by intraperitoneal injection of tumor organoids derived from primary tumors in genetically engineered mice carrying Apc, Kras, Tgfbr2, and Trp53 mutations, and recapitulating human-like metastatic colorectal cancer, we investigated the origin, expansion, and function of peritoneal macrophages during metastatic tumor growth. Our data support that the low inflammatory status of the peritoneal cavity during colorectal cancer peritoneal tumor growth restrains monocyte recruitment and the differentiation of Tim4- resMØs and moMØs while enabling a marked, proliferation-driven expansion of Tim4+ resMØs. Tumor-induced Tim4+ resMØs displayed a migratory and protumor transcriptomic signature characterized by the activation of genes encoding key protumorigenic molecules and potential immunotherapeutic targets, including Adora2a/Adora2b, Arg1, Ido2, Acod1, Mmp12, Cd274, Pdcd1lg2, Spp1, Trem1, and Vegfa. Correspondingly, during peritoneal colorectal cancer tumor growth, Tim4+TREM1+ resMØs migrated to the omentum, the principal peritoneal target organ for metastasis, and promoted colorectal cancer peritoneal tumor progression. These findings may help lead to the development of immunotherapies for colorectal cancer peritoneal metastasis that target tumor-associated peritoneal macrophages.

Peritoneal metastases from colorectal cancer are associated with poor survival and limited therapeutic efficacy. To model patient-specific treatment responses, we generated patient-derived scaffolds (PDS) through tumor decellularization, preserving critical microenvironmental features for functional ex vivo analysis. PDS from 75 metastases were recellularized with HT29 cells and treated with 5-fluorouracil (5-FU), mitomycin C, or oxaliplatin. Gene expression related to proliferation, epithelial phenotype, epithelial-mesenchymal transition, and stemness was quantified and integrated into a composite treatment score to estimate the most favorable therapy for each patient. Treatment responses were influenced by both drug exposure and the patient-specific microenvironment. Importantly, subcluster distributions differed across treatment arms, supporting the concept that therapeutic response is modulated by microenvironmental context rather than being solely drug dependent. These findings were supported in a smaller validation cohort using HCT116 and DLD-1 cells recellularized into selected PDS. Treatment scoring based on HT29 cells revealed heterogeneous sensitivity patterns, with 38.7% of PDS responding best to 5-FU, 36% to mitomycin C, and 25.3% to oxaliplatin. Patients with more than two favorable treatment scores had improved disease-free survival (p = 0.0415). The PDS platform enables functional profiling of chemotherapy-response and may support individualized treatment strategies for colorectal cancer patients with peritoneal metastases.

Receptor-interacting protein kinase 1 (RIPK1) is a key stress sensor regulating cell death, inflammation, and tumorigenesis, yet how RIPK1 becomes activated remains unclear. Here, we identify serine/threonine kinase 38 (STK38) as a novel direct RIPK1 activator. STK38 binds to RIPK1, integrates into RIPK1-containing death complexes, and accelerates RIPK1-dependent cell death. STK38 deletion suppresses RIPK1-mediated necroptosis and apoptosis. Moreover, TNF-α stimulation triggers MEKK2-dependent STK38 activation, which in turn phosphorylates RIPK1 at serine 309, a residue conserved only in higher primates. This phosphorylation at S309 disrupts RIPK1's interaction with its inhibitory kinase MK2, thereby suppressing S320 phosphorylation and facilitating RIPK1 activation. Furthermore, colorectal cancer sample analysis revealed a positive correlation among STK38 expression, RIPK1 activation status, and favourable patient outcomes. Consistently, STK38 deficiency confers resistance to RIPK1-dependent cell death and facilitates tumour progression in a xenograft model. Our findings identify STK38 as an activator of RIPK1 and uncover a novel regulatory mechanism of RIPK1-mediated cell death in humans.

Metastasis remains the leading cause of cancer-related mortality and is driven by pronounced tumour cell plasticity1. Here we identify the transmembrane glycoprotein trophoblast cell-surface antigen 2 (TROP2) as a marker of poor-prognosis colorectal cancer (CRC) associated with WNTlow, fetal-like tumour cell states that are linked to metastasis and therapy resistance. Functional analyses demonstrate that TROP2+ cells exhibit context-dependent stem-like capacity and the ability to initiate metastatic outgrowth. Given that these detrimental tumour states converge on the cell-surface antigen TROP2, we explored therapeutic targeting of this cell population using clinically relevant TROP2-directed antibody-drug conjugates. Time-resolved analyses reveal therapy-associated dynamics in tumour cell state composition between WNThi LGR5+ states and WNTlowTROP2+ fetal-like states. Conventional chemotherapy promotes the induction of TROP2-expressing cells, whereas TROP2 antibody-drug conjugates selectively target these populations and remodel the tumour cell state landscape. Exploiting this plasticity, combined chemotherapy and TROP2 targeting enhances anti-tumour efficacy in patient-derived models. Together, our findings identify TROP2 as a therapeutic vulnerability of CRC and highlight the importance of targeting tumour cell states to improve therapeutic efficacy and overcome resistance in advanced disease.

Patients with colorectal cancer (CRC) frequently develop liver metastases1-3. The prognosis of these patients is skewed by the histopathological heterogeneity of their liver metastases4,5. Patients with 'replacement' metastases have a 5-year overall survival of less than 44.2%, compared with 73.4% in patients with 'encapsulated' (previously known as desmoplastic) metastases5; yet there are currently no approved therapies targeting replacement liver metastases. Here we show that treatment-naive patients with CRC with liver steatosis have an increased occurrence of replacement metastases compared with patients without steatosis. Mechanistically, we find that steatosis-promoted fatty acid oxidation increases formation of replacement metastases by increasing MYC stability through acetylation. In turn, MYC activates proline synthesis, fuelling collagen production, enabling growth of replacement metastases. Targeting MYC, P5CS or COL1A1 suppresses the occurrence and growth of replacement metastases in patient-derived organoids, mouse or patient-derived xenograft models. Spatial metabolite and protein analyses of liver metastases from patients with CRC further support this mechanism. In conclusion, we provide a mechanistic understanding of the emergence of liver metastases with poor prognosis in treatment-naive patients with CRC, identifying potential targets for therapeutic intervention.

Metastasis, responsible for > 90% of cancer-related mortality, represents the most lethal yet least mechanistically understood phase of cancer progression. A critical bottleneck is tumor cell migration through physically confined environments, including dense extracellular matrix, narrow capillaries and endothelial gaps. Although tumor cells reprogram their metabolism to facilitate cancer progression, it remains unclear how specific metabolic adaptations enable them to overcome the unique physical challenges posed by these confined spaces, thereby promoting distant metastasis. We conducted a CRISPR screen targeting 1685 metabolic enzymes and identified dihydrolipoamide dehydrogenase (DLD), a mitochondrial enzyme involved in energy metabolism, as essential for confined migration of tumor cells. Depletion or pharmacological inhibition of DLD suppressed CRC metastasis by impairing tumor cell migration through capillaries and endothelial gaps. Upon mechanical compression, heterogeneous nuclear ribonucleoprotein A0 (hnRNPA0) binds to the adenylate uridylate-rich element (ARE) in the 3'UTR of DLD, enhancing its mRNA stability and upregulating DLD expression in tumor cells during confined migration. Elevated DLD expression enhances tricarboxylic acid (TCA) cycle metabolism, increasing malate levels. Malate interacts with tubulin alpha-1B chain (TUBA1B) to promote microtubule assembly, facilitating confined migration and metastasis. Knock-in of an ARE-deleted DLD mutant (DLD ΔARE) or disruption of the malate-TUBA1B interaction significantly suppressed tumor metastasis. In CRC patients, DLD expression was upregulated in tumor cells within capillaries of primary tumors and correlated with metastatic recurrence. Our findings reveal that compressive forces drive metastatic dissemination by epigenetically reprogramming mitochondrial metabolism, which in turn fuels cytoskeletal remodeling.

Colitis-associated colorectal cancer (CAC) is a serious complication of inflammatory bowel disease. As stress-responsive membraneless organelles, stress granules (SGs) are involved in modulating inflammatory suppression and promoting tumorigenesis, but their role in the pathogenesis of CAC remains unclear. This study reveals that SGs exhibit a stage-specific function during CAC progression, being protective in acute colitis but transitioning to a tumor-promoting role in the dysplasia-carcinoma sequence. We first demonstrate that, during acute colitis, fluctuating oxidative stress drives dynamic SGs assembly. However, despite persistently elevated oxidative stress from chronic colitis to dysplasia, SGs levels paradoxically decline. This decline is attributed to the upregulation of TOM1, a novel negative regulator that binds the NTF2L domain of the core SGs scaffold protein G3BP1 to promote SGs disassembly, thereby counteracting the oxidative stress-driven assembly. The consequent disassembly facilitates the nuclear translocation of the oncogenic transcription factor FUBP1, which in turn promotes c-Myc expression and thereby contributes to tumorigenesis. Our findings establish the regulation of SGs dynamics, particularly via TOM1 and G3BP1, as a promising therapeutic strategy for CAC.

Tissue classification is one of the fundamental tasks in computational pathology, but domain shifts in digital pathology images limit the generalization of classification models. Domain generalization has emerged as a leading solution to address this gap, with related research often using multiple public datasets to demonstrate model generalization ability across different sources. To further explore this, we introduce the GDPH-CRC-HE-MS dataset, consisting of 101 H&E-stained colorectal cancer slides from Guangdong Provincial People's Hospital, scanned by 1 to 6 different scanners. In this study, we propose an unsupervised single-domain progressive generalization (USD-PG) framework, which incorporates two progressive data transformations: style progressive data transformation (Style-PDT) and spatial progressive data transformation (Spatial-PDT). This approach prevents unreasonable texture and color changes caused by completely random transformations during the early training stages. We evaluate the generalization ability of the USD-PG framework on the new GDPH-CRC-HE-MS dataset as well as the publicly available NCT-CRC-HE-100K dataset. Our results demonstrate that USD-PG achieves superior performance in single-source domain generalization for tissue classification, effectively handling both scanner-based and data-source domain shifts. It highlights the potential of USD-PG for enhancing domain generalization in tissue classification and its applicability in clinical settings. The source code and the released datasets are available at: https://github.com/linjiatai/USD-PG.

The human gut microbiome is shaped by diverse selective forces that originate from host and environmental factors and it substantially influences health and disease. Whereas the association of microbial lineages with various health conditions has been shown at different taxonomic levels1-5, the extent to which unifying adaptive mechanisms sort microbial lineages into ecologically differentiated populations remains poorly understood. Here we show that genome-wide selective sweeps are a pervasive mechanism that differentiates bacteria in the microbiome. This mechanism leads to population structures akin to global epidemics across geographically and ethnically diverse human populations. Such sweeps arise when an adaptation allows a clone to outcompete others in its niche followed by rediversification, and they manifest as clusters of closely related genomes on long branches in phylogenetic trees. This structure is revealed by excluding recombination events that mask the clonal descent of the genomes. Indeed, we show that genome-wide sweeps originate under a wide range of recombination rates in at least 66 taxa from 25 bacterial families. Estimated ages of divergence suggest that sweep clusters can spread globally within decades and that this process has occurred throughout human history. Sweep clusters are associated with different host conditions-such as age, colorectal cancer, inflammatory bowel diseases and type 2 diabetes-as an indication of their ecological differentiation. Our results reveal an evolutionary mechanism for the observation of stably inherited strains with differential associations and provide a theoretical foundation for analysing adaptation among microbial populations.

The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway represents a central driver of innate immune activation, and manganese ion (Mn2+) has recently been identified as a potent modulator of this signaling axis. However, the application of Mn2+ is limited by its rapid clearance, nonspecific distribution and potential neurotoxicity. Inspired by the unique chemical structure and biological functions of adenosine triphosphate (ATP), we herein propose an ATP-Mn coordination nanoparticle (ATP-Mn CNP) to fuel cGAS-STING activation and antitumor immunity. We demonstrated that the phosphate groups of ATP could coordinate with Mn2+ and form stable, well-defined nanoparticles after lipid coating. ATP-Mn CNP significantly increased the expression of cGAS-STING-associated genes and activated the corresponding signaling cascades, and thus effectively polarized macrophages from tumor-supportive M2 to antitumor M1 phenotype. In vivo antitumor studies indicated that ATP-Mn CNP treatment significantly suppressed tumor growth, and reprogramed macrophages in tumors and draining lymph nodes, which thus facilitated the tumor infiltration of cytotoxic lymphocytes. Combination of ATP-Mn CNP with immune checkpoint inhibitors achieved 37.5% tumor eradication in MC38 murine models, and significantly prolonged mice survival. This study establishes an ATP-fueled coordination strategy that harnesses ATP as both an assembly ligand and an immune stimulator to enhance Mn-mediated STING activation.

Hypoxia within solid tumors has been identified as one of the main obstacles in radiotherapy due to the severely reduced radiosensitivity. Current strategies to alleviate tumor hypoxia mainly rely on oxygen supplementation using oxygen carriers (e.g., hemoglobin- or perfluorocarbon-based systems), hypoxia-activated prodrugs, or tumor oxygen consumption modulators, leading to limited efficacies due to poor tumor-specific targeting, insufficient oxygen delivery in the complex tumor microenvironment, and potential systemic toxicity. Here we propose an alternative but novel strategy for radiosensitization in colorectal cancer radiotherapy by uncompacting the tumor tissue via tumor disaggregation, thus alleviating tumor hypoxia and enhancing radiosensitivity. This strategy has been realized by developing a nanomedicine composed of ethylene diamine tetraacetic acid-loaded layered double hydroxide (LDH/EDTA) featuring intratumoral acidity-responsive EDTA release. The released EDTA deprives Ca2+ ions from the intercellular cadherins that connect tumor cells through EDTA- Ca2+ chelation, thus disrupting the inter-cellular junctions in tumor tissue by cadherin damages. As a result, compactness and rigidity of tumor tissues are greatly reduced, and the ambient oxygen is allowed to diffuse deep into the tumor interior, thereby alleviating the hypoxia of solid tumors and effectively enhancing their sensitivity to radiotherapy. This work proposes a novel yet facile strategy to enhance radiosensitivity simply by overcoming the physical barriers of tumors and alleviating hypoxia.

Berberine (BBR), the predominant isoquinoline alkaloid in Coptidis Rhizoma, exhibits remarkable anti- colorectal cancer (CRC) activity. However, whether BBR triggers CRC cell death through ferroptosis-associated disruption of energy metabolism remains to be elucidated. To investigate if BBR induces mitochondrial energy metabolism disorder in CRC cells by regulating the ferroptosis signaling pathway. BBR's effects on malignant phenotypes were evaluated in vitro (human cell line HCT116, murine cell line CT26 cells at 10, 20, 40 μM) and in vivo (80 mg/kg). Target engagement and mechanistic pathways were interrogated through RNA-sequence combined with convolutional neural network-based pathway prediction, corroborated by surface plasmon resonance, cellular thermal shift assay. Downstream validation mainly included quantification of Gli1, STAT3, GPX4, SLC7A11, and FTH1 expression via RT-qPCR, Western blot, immunofluorescence, and other molecular expression and functional confirmation experiments. BBR inhibited CRC cell proliferation with IC50 value for HCT116 cells for 48 h at 19.86 ± 2.31 μM, and for CT26 cells for 48 h at 21.35 ± 2.63 μM. Concurrently, it elevated ferroptosis markers such as malondialdehyde, lactate dehydrogenase, Fe2+, and 4-hydroxynonenal, while suppressing ATP levels, superoxide dismutase activity, and energy metabolism-related enzymes. Graph convolutional network-based drug "on-target" pathway algorithm predicted Gli1 as top-9 target, and surface plasmon resonance confirmed direct BBR-Gli1 binding with KD value at 0.652 μM, cellular stability thermal assessment showed BBR stabilized Gli1 with thermal shift with ΔT = 2.3 °C. Mechanistically, BBR exerted its anti-CRC effects by inhibiting the Gli1/STAT3-ferroptosis negative regulation (Gli1/STAT3-FNR) axis, a novel regulatory pathway. Notably, BBR exhibited no significant organ or hematological toxicity in vivo at the experimental doses. BBR triggers ferroptosis-mediated energy metabolism disorder by inhibiting Gli1/STAT3-FNR axis. This work provides a mechanistic support for BBR anti-CRC indications, and suggests an encouraging approach for treating CRC.

Disruptions in rhythm-regulating genes, termed circadian misalignment, have been increasingly associated with the progression of colorectal cancer (CRC), a significant global health challenge. Capsaicin (CAP), a bioactive compound found in chili pepper, has emerged as a promising modulator of clock genes, exhibiting anti-CRC potential. We propose a previously unrecognized dual-pathway mechanism where CAP may modulate circadian clock genes via both transient receptor potential vanilloid 1 (TRPV1)-dependent and TRPV1-independent pathways to inhibit colorectal carcinogenesis. Although the role of CAP in CRC prevention via clock gene regulation is compelling, direct mechanistic evidence remains limited. This paper also identifies several research gaps to underscore the necessity for further research to unlock its therapeutic potential and develop novel preventive strategies.

The dominance of immunotherapy-insensitive MSS colorectal cancers (CRCs), which represent most cases, contrasts sharply with the treatable MSI-H minority, making this disparity a key obstacle to progress. It is urgent to identify genes driving immune evasion in MSS CRCs. Here, using a genome-wide CRISPR screen in a syngeneic tumor model under immune pressure, we identify TMEM184A as a previously unknown tumor-intrinsic regulator of immune evasion. Its genetic deletion in murine models enhanced CD8+ T cell infiltration and increased surface MHC-I expression on cancer cells, as shown by flow cytometry, immunohistochemistry, immunofluorescence and RNA-seq. Mechanistically, TMEM184A functions as a novel macroautophagy/autophagy receptor by binding GABARAPL2, directly promoting the autophagic degradation of IFNG-induced MHC-I. In murine models, genetic deletion of Tmem184a led to a significant increase in both CD8+ T cell infiltration and surface MHC-I expression on cancer cells. Functional studies in vivo and in vitro confirmed that this impaired antigen presentation causally facilitates immune evasion. Our findings establish MHC-I autophagic degradation as a critical pathway regulating immune evasion and position TMEM184A as a pivotal molecular hub in this process. Notably, treatment with the autophagy inhibitor chloroquine significantly increased surface MHC-I levels and enhanced the efficacy of anti-PDCD1/PD-1 therapy specifically in TMEM184A-high tumors. This work suggests that targeting TMEM184A or its associated autophagic pathway could restore antigen presentation in MHC-I-deficient tumors, offering a potential combinatorial strategy to overcome adaptive immune resistance in multiple malignancies.Abbreviations: AKP organoids:apcandtrp53knockout, KRASG12Dmutation organoids; CRC: colorectal cancer; CQ: chloroquine; GABARAPL2/Atg8: GABA type A receptor associated protein like 2; IF: immunofluorescence; IFNG: interferon gamma; IHC: immunohistochemistry; MHC-I: major histocompatibility complex I; qRT-PCR: quantitative reverse transcription PCR; MSI-H: microsatellite instability-high; MSS: microsatellite-stable; TMEM184A: transmembrane proteins 184a.

Immune checkpoint blockade targeting the PD-1/PD-L1 axis shows limited efficacy in microsatellite-stable (MSS) colorectal cancer (CRC), primarily due to an immunosuppressive tumor microenvironment (TME) and insufficient T-cell activation. Here, we report a peptide coacervate-mediated siRNA delivery platform that enables coordinated gene silencing of PD-1 in T-cells and PD-L1 in tumor cells to enhance CRC immunotherapy. HBpep-SP coacervates (HCs) were functionalized with anti-CD3 antibodies to generate targeted coacervates (THCs), enabling efficient T-cell-targeted delivery of PD-1 siRNA, robust PD-1 knockdown, and enhanced T-cell effector function, as indicated by increased IL-2 and IFN-γ production. In parallel, HCs efficiently delivered PD-L1 siRNA into CRC cells, achieving significant PD-L1 knockdown. Dual checkpoint silencing in a co-culture system of T-cells and CRC cells synergistically enhanced T-cell proliferation and activation, leading to increased tumor cell apoptosis. Importantly, in a murine MSS CRC model, intratumoral co-administration of siPD-1@THC and siPD-L1@HC simultaneously suppressed PD-1 and PD-L1 expression within the TME, increased intratumoral T-cell abundance, and elevated pro-inflammatory cytokine levels, resulting in restored antitumor immunity and significant tumor growth inhibition. Collectively, this peptide coacervate-based dual-checkpoint RNA interference strategy provides a promising approach for advancing T-cell-mediated immunotherapy in MSS colorectal cancer.

Oral administration can deliver targeted nanoparticles directly to colorectal cancer (CRC) lesions, bypassing systemic circulation. However, orally administered nanoparticles may become physically entrapped in the small intestine, compromising therapeutic efficacy. To overcome this limitation, we designed and prepared folic acid-modified, CRC cell-targeting nanoparticles (FA/CS/TPP/β-acids) and incorporated them into a sodium alginate hydrogel (Alg/FA/CS/TPP/β-acids). The release of β-acids from the nanoparticles was pH-dependent, with cumulative release following the order: pH 7.8 > pH 6.8 > pH 1.2. The nanoparticle/hydrogel system exhibits good biocompatibility (hemolytic activity < 5%; non-toxic to NCM460 cells) and significantly enhances cellular uptake in HCT116 and HT29 cells. In vivo studies showed that the nanoparticles were non-toxic and effectively inhibited CRC. Interestingly, the hydrogel undergoes a sol-gel transition under acidic gastric conditions (pH 1.2). Upon reaching the colorectum, where the pH is higher, it undergoes a gel-sol transition. These results demonstrate that the alginate-modified hydrogel can protect the nanoparticles in the gastrointestinal environment, enabling targeted release for CRC therapy.

Secondhand smoke exposure (SHSE) poses risks to human health, yet the evidence remains fragmented. To address this, we conducted an umbrella review (Registration No. CRD42024583424 ) to synthesize and evaluate epidemiological evidence across the lifespan. We systematically searched four databases: PubMed, the Cochrane Library, Web of Science and Embase up to 24 August 2025, for systematic reviews with meta-analyses of observational studies that examined any health outcome in relation to SHSE. Methodological quality was assessed using AMSTAR 2, while evidence credibility and robustness were evaluated through evidence classification criteria, fail-safe number and GRADE framework. A total of 111 systematic reviews with meta-analyses involving approximately 253.3 million participants, encompassing 130 health outcomes and 226 independent associations across developmental stages from fetal development and childhood to adulthood, were included in the reanalysis. Convincing evidence demonstrated a dose-dependent association between prenatal maternal smoking and adverse pregnancy outcomes, such as infant low birth weight (11-19 cigarettes per day, odds ratio [OR] = 2.05, 95% confidence interval [CI] 1.89-2.23, k = 7, p < 0.001). Highly suggestive evidence also showed associations between SHSE and lung cancer (relative risk [RR] = 1.24, 95% CI 1.16-1.32, k = 82, p < 0.001), cardiovascular diseases (OR = 1.24, 95% CI 1.16-1.31, k = 52, p < 0.001) and depressive symptoms (OR = 1.39, 95% CI 1.28-1.51, k = 24, p < 0.001) with evidence of dose-response relationships. Despite limitations of observational data, this review distinguishes evidence saturation areas from key research gaps, underscores health risks especially for pregnant women and children, and may inform tobacco control interventions.

Dirofilaria nematodes, a common cause of canine filarial disease, are increasingly recognized as emerging human pathogens. We report a case of human pulmonary dirofilariasis in the lung of a man from Northern Australia with pulmonary adenocarcinoma. This case highlights the risk for zoonotic transmission in regions with high canine heartworm prevalence.

The anti-PD-1, anti-VEGF bispecific antibody ivonescimab plus chemotherapy significantly improves overall survival in advanced squamous non-small cell lung cancer versus an immunotherapy-chemotherapy combination, according to a trial conducted in China. However, questions remain over the trial's applicability to patients in other countries.

Results from LIBRETTO-432 demonstrate that adjuvant selpercatinib yields a substantial improvement in event-free survival for patients with early-stage RET fusion-positive non-small cell lung cancer. Experts say the findings establish selpercatinib as a new standard of care for this rare disease, although questions remain over identifying patients and access to screening.

Findings from the phase III WU-KONG28 trial indicate that the next-generation tyrosine kinase inhibitor sunvozertinib, an approved later-line option for patients with non-small cell lung cancer harboring EGFR exon 20 insertions, also looks effective up front, besting chemotherapy. Meanwhile, updated CHRYSALIS-2 data point to amivantamab combined with lazertinib as a new option for patients with atypical EGFR mutations, with initial efficacy translating to durable overall survival.

Epidermal growth factor receptor (EGFR) exon 20 insertions are the third most common EGFR mutation subtype in non-small cell lung cancer, occurring in up to ~4% of cases. Although the clinical profile overlaps with classical sensitizing EGFR mutations, most exon 20 insertions confer primary resistance to earlier generation epidermal growth factor receptor tyrosine kinase inhibitors, reflecting position- and variant-dependent steric constraints within the adenosine triphosphate-binding pocket. More than 100 distinct variants have been described, motivating structure- and variant-informed frameworks for diagnosis and treatment selection. Accurate detection is therefore essential: fixed-content polymerase chain reaction assays may miss clinically actionable variants, whereas next-generation sequencing improves variant capture and supports standardized reporting, including identification of co-alterations. Plasma cell-free DNA testing can complement tissue profiling when biopsy is not feasible, but low circulating tumour DNA shedding can yield false-negative results, supporting reflex tissue testing after negative plasma findings when clinical suspicion remains high. Therapeutically, the landscape has shifted rapidly since 2021, with the introduction of exon 20 insertion-directed strategies including bispecific antibodies and next-generation tyrosine kinase inhibitors, alongside an expanding pipeline of mutant-selective agents and rational combinations. Despite these advances, response durability remains limited for many patients, and both intrinsic and acquired resistance via on-target changes, bypass signalling and tumour evolution continues to drive unmet need. Future progress will likely depend on central nervous system-active strategies, adaptive sequencing guided by longitudinal molecular profiling and combination approaches that address pathway bypass and tumour heterogeneity.

Antibody-drug conjugates (ADCs) are complex molecules composed of a monoclonal antibody, a linker and a cytotoxic payload. Their design enables the selective delivery of cytotoxic agents to tumoral cells through antibody binding to a tumor-expressed antigen, followed by internalization, intracellular degradation and payload release, ultimately enabling the cytotoxic drug to exert its antitumor activity. ADCs have been evaluated in phase II and III trials in previously treated advanced non-small cell lung cancer (NSCLC), both in oncogene-addicted and in non-oncogene-addicted tumors, addressing resistance to standard therapies. Ongoing clinical trials are now expanding their use both in the first-line setting for advanced disease and in earlier disease stages. This narrative review summarizes the currently available data for ADC treatment in NSCLC, highlighting the need for improved patient selection to maximize benefit while limiting toxicity, incorporating clinical characteristics, pharmacogenomics and optimal treatment sequencing in the equation. Moreover, the understanding of resistance mechanisms and the development and validation of predictive biomarkers will be of utmost relevance to inform clinical practice.

Adherence to subsequent lung cancer screening (LCS) is essential but not well characterized in biennial national programs. To assess whether baseline LCS results and their management are associated with subsequent screening adherence and clinical outcomes. We analyzed participants in the Korean national LCS program who underwent baseline low-dose computed tomography (CT) between 2019 and 2021 and remained lung cancer-free for 2 years. Baseline results were classified as true negative or false positive (FP); FPs were grouped by management (invasive diagnostic procedures, chest CT surveillance, or no further evaluation). Adherence to the next biennial screening was modeled using multivariable logistic regression. Lung cancer incidence and all-cause mortality were assessed using Cox models with a 2-year landmark design. Among 235,753 participants, 54.4% returned for subsequent screening. Compared with true-negative results, adherence was lower among those with FP results who underwent invasive procedures (adjusted odds ratio [aOR], 0.64; 95% confidence interval [CI], 0.56-0.72) or CT surveillance (aOR, 0.77; 95% CI, 0.74-0.80) but was similar among those with FP results without further evaluation (aOR, 1.00; 95% CI, 0.95-1.04). In the landmark analysis, FPs were associated with higher risks of lung cancer incidence (adjusted hazard ratios [aHRs], 2.95-2.49 across management groups) and higher all-cause mortality among those who underwent invasive procedures (aHR, 1.57; 95% CI, 1.10-2.24) or CT surveillance (aHR, 1.23; 95% CI, 1.07-1.41). Downstream management after baseline FP findings, rather than FP status alone, was associated with subsequent LCS adherence, highlighting the need to support return screening in higher risk FP groups.

Postoperative pulmonary complications are common after lung cancer surgery in older adults. Individualised positive end-expiratory pressure may optimise intra-operative lung mechanics, but its effect on postoperative pulmonary complications is uncertain. We hypothesised that individualised positive end-expiratory pressure would reduce the incidence of postoperative pulmonary complications compared with a fixed positive end-expiratory pressure in older patients (age ≥ 60 years) undergoing lung cancer surgery. In total, 400 patients were allocated randomly to individualised positive end-expiratory pressure (PEEPIND group) or a fixed positive end-expiratory pressure of 5 cmH2O (PEEP5 group). The primary outcome was the incidence of postoperative pulmonary complications. Secondary outcomes included duration of postoperative hospital stay; extrapulmonary complications; 30-day postoperative complications; driving pressure; and oxygenation index. Median (IQR [range]) individualised positive end-expiratory pressure was 11 (9-11 [3-13]) cmH2O during one-lung and 9 (7-9 [3-13]) cmH2O during two-lung ventilation. Patients allocated to the PEEPIND group had lower driving pressures during one-lung (12 (11-14 [4-23]) vs. 15 (13-18 [7-24]) cmH2O, p < 0.001) and two-lung ventilation (9 (7-13 [4-26]) vs. 12 (10-14 [5-26]) cmH2O, p < 0.001) and a higher oxygenation index during one-lung ventilation (26.7 (20.3-34.4 [6.7-55.9]) vs. 22.7 (16.0-29.3 [8.8-58.9]) kPa, p < 0.001) compared with those allocated to the PEEP5 group. Despite this, the incidence of postoperative pulmonary complications was similar between groups (PEEPIND group 54/195 (28%) vs. PEEP5 group 50/197 (25%), risk ratio 1.09, 95%CI 0.79-1.52, p = 0.60). Electrical impedance tomography-guided individualised positive end-expiratory pressure reduced driving pressures and improved intra-operative oxygenation but did not decrease the incidence of postoperative pulmonary complications in older adults undergoing lung cancer surgery.

A 72-year-old woman was appointed for 18 F-FDG PET/CT for restaging of lung adenocarcinoma. Initial whole-body 18 F-FDG PET/CT revealed a focal hypermetabolic area in the right lung, suspicious for lung metastasis, while no abnormalities were present in the partially depicted skull. Since the patient presented with a massive headache during examination, an additionally performed cranial PET/CT scan without further tracer injection revealed an intracerebral hematoma and wedge-shaped glucose hypometabolism in the left occipital lobe. An additional contrast-enhanced cranial CT performed 45 minutes after cranial PET/CT scan revealed progressive hematoma of the complete left hemisphere, mid-line shift, and tentorial herniation.

Racial and ethnic disparities are a concern in lung cancer screening. To investigate the performance of risk prediction models to define screening eligibility across 4 U.S. racial and ethnic groups. Cohort study. United States, Lung Cancer Cohort Consortium. 641 830 participants aged 50 to 80 years with a smoking history from 12 U.S. cohorts, including 6390 Asian, 9781 Hispanic, 39 872 non-Hispanic Black, and 585 787 non-Hispanic White participants. Calibration and discrimination were quantified for 16 lung cancer prediction models. Then, screening-related metrics were calculated after applying model thresholds to select the same number of eligible participants as the 2021 criteria from the U.S. Preventive Services Task Force (USPSTF-2021). These included eligibility, sensitivity, and efficiency measured as estimated number needed to screen (NNS; the ratio between participants and lung cancer cases) for each strategy or prediction model in each racial and ethnic group. General patterns across the 16 models included substantial underestimation of lung cancer risk in non-Hispanic Black participants (expected-observed ratio < 0.75 for 11 of 16 models), lower discrimination in Asian participants than all other groups (13 of 16 models), and lower discrimination in non-Hispanic Black than non-Hispanic White participants (15 of 16 models). When a same-sized screening-eligible population as USPSTF-2021 (38.0%) was enforced, all risk-based strategies achieved better average estimated screening efficiency and reduced racial and ethnic differences in efficiency compared with USPSTF-2021. The Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial Model 2012 (PLCOm2012) and Life Years gained From Screening-Computed Tomography model (LYFS-CT) performed best (mean estimated NNS, 36.5 [SD, 8.8] and 40.1 [SD, 8.2], respectively). However, no strategy could simultaneously optimize eligibility, sensitivity, and efficiency while also reducing racial and ethnic differences. Smaller sample for Asian and Hispanic participants. To optimize efficiency and minimize its variation across racial and ethnic groups, risk-based strategies were superior to USPSTF criteria. Further optimization of prediction models for the diverse U.S. population is needed. U.S. National Cancer Institute, Lung Cancer Research Foundation, and Cancer Research UK.

We describe a 71‑year‑old man with squamous cell lung carcinoma who underwent metastasis work-up with CT scan and 99mTc‑MDP bone scintigraphy, which suggested limited extrathoracic skeletal involvement. Subsequent 99mTc‑FAPI‑46 scintigraphy revealed a substantially greater disease burden, identifying a more extensive nodal and skeletal metastases. These findings resulted in a decisive stage migration from an initially presumed oligometastatic to a clearly polymetastatic stage IV disease, with direct implications for therapeutic intent. This case underscores the potential clinical utility of 99mTc‑FAPI‑46 scan as a cost‑effective staging modality, particularly in settings where PET/CT is unavailable or financially prohibitive.

Pembrolizumab 400 mg every 6 weeks (Q6W) was approved based on population pharmacokinetic (pop-PK) modeling rather than pharmacokinetic measurements, leaving real-world concentrations insufficiently characterized. The impact of concentration changes when switching from 200 mg every 3 weeks (Q3W) to Q6W on immune-related adverse events (irAEs) remains unclear. This study aimed to evaluate the association between the change in pembrolizumab dosage and irAE occurrence. We retrospectively identified patients with advanced non-small cell lung cancer who switched pembrolizumab from Q3W to Q6W between March 2017 and September 2023. Serum concentrations were quantified; pop-PK analysis estimated maximum concentration (eCmax) and estimated trough exposure (eCtrough). Intercycle variability was expressed as percentage coefficient of variation (%CV) using all available 200 mg Q3W cycles. Early irAEs were defined as events within 126 days.Associations between pop-PK metrics and irAEs (including pneumonitis) after switching were explored. Clinical correlates of %CV-eCmax were also assessed. 79 patients switched, and pop-PK analysis was feasible in 66 patients. The median age was 70 years, 50 were male, and 62 had a smoking history. The median Q3W cycles were 6, and the median follow-up period after switching was 463 days. After switching, 45 events occurred new or worsened irAEs, with 19 pneumonitis, and 22 had early irAEs, with 13 early pneumonitis.Among the 64 patients who received ≥2 cycles of Q3W, those who developed irAEs after switching showed higher %CV-eCmax during Q3W. Similar associations were observed for early irAEs, pneumonitis, and early pneumonitis.%CV-eCtrough during Q3W was not associated with irAEs after switching, with an association observed only for early pneumonitis.High %CV-eCmax during Q3W was associated with increased odds of irAEs, pneumonitis, early irAEs, and early pneumonitis. No significant associations were observed between %CV-eCtrough and irAEs or early irAEs.Higher platelet count and renal dysfunction were independently associated with increased %CV-eCmax. High intercycle eCmax variability during 200 mg Q3W is associated with an increased risk of new or worsened irAEs, particularly pneumonitis, after switching to 400 mg Q6W, suggesting that intercycle variation in Cmax may be an important safety consideration when switching to 400 mg Q6W.

Tumor heterogeneity-encompassing diverse cellular phenotypes, genomic alterations, and microenvironmental contexts-is a principal barrier to effective cancer therapy. Single-cell RNA sequencing (scRNA-seq) has transformed our ability to resolve this complexity by capturing transcriptomes at single-cell resolution. Here, we review the technical foundations required for high-quality scRNA-seq studies. We then trace the evolution of scRNA-seq platforms from manual micromanipulation to high-throughput systems, and describe the computational pipelines that enable reliable data interpretation. The application of scRNA-seq is exemplarily shown in the context of lung cancer, where single-cell profiling has revealed (i) the clonal and sub-clonal architecture of tumors, (ii) extensive remodeling of the immune microenvironment, iii) key mechanisms underlying resistance to targeted agents and immune-checkpoint blockade, and (iv) the dynamics of neo-antigen-specific T-cell responses. Integrating machine-learning techniques-such as deep-learning classifiers and graph-based models-with single-cell transcriptomic data has markedly sped up biomarker discovery, produced more accurate risk-stratification scores, and enabled the generation of patient-specific therapeutic predictions. We surveyed the major trial registry ClinicalTrials.gov and identified ∼380 ongoing or completed studies that explicitly incorporate scRNA-seq as a correlative or pharmacodynamic endpoint. Overall, the analysis shows that scRNA-seq becomes an increasingly important component of modern trials, providing high-resolution cellular and molecular readouts that complement conventional imaging and bulk-omics endpoints. While key challenges remain, ranging from costs, scalability and need for rigorous validation before routine clinical deployment, ongoing technological advances continue to expand the potential of scRNA-seq as a cornerstone of precision medicine.

Polygenic risk scores (PRSs) quantify genetic susceptibilities, yet ancestry imbalance in genome-wide association studies (GWASs) limits the accuracy of monoracial PRSs in non-European populations. Here, we perform a multiancestry GWAS meta-analysis for lung cancer (76,953 cases and 1,886,372 controls), identifying 87 conditionally independent genome-wide significant loci, including two unreported cytobands. We use a PRS construction method, PRS-CSx, to develop a multiancestry PRS (PRSMA) which outperforms 32 published PRSs. To enhance predictive power, we construct a multitrait PRS (PRSMT) using CatBoost, integrating 32 cross-trait PRSs across three ancestries. Combining PRSMA and PRSMT, we generate PRSMAMT and validate it in independent cohorts (OncoArray, TRICL and All of Us). PRSMAMT demonstrates superior discriminability in European, Asian, and African populations, improves risk stratification, and identifies approximately 10% additional lung cancer cases in the UK Biobank. Individuals with elevated PLCOm2012 scores and high genetic risk exhibit a 12.64-fold higher cumulative risk than those with low scores and low genetic risk, supporting precision prevention strategies.

Non-small-cell lung cancer (NSCLC), the predominant type of lung cancer, is characterized by high invasiveness and significant mortality. Despite its clinical impact, the molecular mechanisms driving its pathogenesis and progression remain poorly understood. This study demonstrates that TMED9 is overexpressed in NSCLC and showed using multiple independent sample sets that its expression level is significantly associated with poor patient prognosis. Gain- and loss-of-function experiments revealed that TMED9 promotes proliferation, invasion, and migration of NSCLC cells in vitro and significantly accelerates tumor growth and metastasis in vivo. Mechanistically, TMED9 interacts with ATG9A and recruits USP5 to facilitate the deubiquitination and stabilization of ATG9A, thereby activating autophagy and driving malignant progression. Notably, genetic depletion of TMED9 enhances the sensitivity of NSCLC cells to osimertinib. Collectively, these findings identify the TMED9-USP5-ATG9A signaling axis as a critical driver of NSCLC malignancy, highlighting TMED9 as a promising therapeutic target.

Liposomes, as one of the most established nanocarriers, have been widely employed in lung cancer therapy, which remains the leading cause of cancer-related mortality worldwide. However, the current landscape of liposomal formulations and the principles guiding their rational design remain insufficiently defined. This review provides a comprehensive analysis of how liposomal delivery systems have been engineered to address biological and therapeutic challenges in lung cancer. We analyze liposome-based delivery of chemotherapeutic agents, including formulations that have advanced to clinical trials, with particular emphasis on strategies developed to overcome multidrug resistance (MDR). Surface engineering and functionalization strategies that enable active targeting and enhance therapeutic efficacy are also reviewed. Liposomal platforms for immunotherapy are examined, alongside their application in delivering phytochemicals, nucleic acid therapeutics, and photo-enabled modalities, including photothermal and photodynamic therapies, which are discussed in dedicated sections. Cutting-edge innovations, such as biomimetic liposomes, stimuli-responsive smart systems, and theranostic designs integrating diagnostic and therapeutic functions, are highlighted. Finally, pulmonary delivery of anticancer agents is evaluated, focusing on formulation parameters informed by current liposome-based evidence and broader lessons from nanomedicine. By providing mechanistic insights, design principles, and translational perspectives, this review offers a unified framework for liposome-based platforms in lung cancer therapy.

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are considered as classic targeted drugs for EGFR-mutated non-small cell lung cancer (NSCLC), but induce EGFR-TKI-resistance and immunosuppression at advanced stage. The targeted inhibition of YES-associated proteins (YAP) combined with immunomodulation is expected to be a distinctive supplementary approach for EGFR-TKI-resistant NSCLC therapy. Herein an endoplasmic reticulum (ER)-targeting Zn/Cu-bi-single-atom nanoplatform (Zn/Cu-BSRGT) was prepared for EGFR-TKI resistance reversion, cascaded ER stress and immunoactivation. Specifically, after precisely targeting to the ER, Zn/Cu-BSRGT NPs provided simultaneous release of ·OH and 1O2 through the efficient chemodynamic therapy (CDT) and sonodynamic therapy (SDT), triggering intense ER stress. Meanwhile, the released YAP-siRNA interfered with the expression of YAP and the EGFR bypass signaling pathway, reversing the AXL-mediated resistance to EGFR-TKI. Furthermore, significant glucose consumption and ER stress triggered the immunogenic cell death (ICD) and systemic immune activation, and down-regulated the PERK-Nrf2 signaling pathway and multidrug resistance protein (MRP1). In summary, the combined application of single-atom-nanozyme catalytic technology and gene-targeted silencing technology successfully reversed EGFR-TKI resistance and promoted immunoactivation in NSCLC under ER-targeting assistance, providing support for the new strategic development of drug-resistant NSCLC.

While radiotherapy (RT) is effective for local tumor control, it rarely induces the regression of non-irradiated metastases, a phenomenon known as the abscopal effect. The mechanisms constraining this systemic immune response remain poorly understood. This study investigates galectin-9 (Gal-9), a ligand for the TIM-3 (T-cell immunoglobulin and mucin-domain containing-3) immune checkpoint, as a mediator of immune escape that limits RT systemic efficacy and evaluates combinatorial RT/Gal-9 blockade in preclinical models. METHODS: Multiplatform analysis (RNA sequencing, immunoblot, flow cytometry, ELISA, immunohistochemistry) characterized RT-induced Gal-9 regulation in human lung/colorectal cancer cell lines, murine tumors/serums, and paired patient tumors. Local and abscopal therapeutic efficacy was evaluated in homologous (CT26/CT26, LLC/LLC) and heterologous (CT26/4T1) two-tumor mouse models. Immune profiling of tumor microenvironment, tumor-draining lymph nodes (tdLNs), and splenic compartments was comprehensively assessed by flow cytometry. Mechanistic studies employed STING (Stimulator of Interferon Genes) inhibition (H151), CD8+ T-cell depletion (anti-CD8α), macrophage/monocyte depletion (PLX-3397), interferon-I (IFN-I) blockade (anti-IFNAR1), and lymphocyte egress inhibition (FTY720). RT upregulated Gal-9 predominantly within host myeloid compartments (dendritic cells, macrophages, monocytes, neutrophils) versus tumor cells, in both irradiated tumors and abscopal tumors. Mechanistically, RT-activated STING-IFN-I axis locally induced Gal-9+ myeloid cells that subsequently disseminated systemically. Clinically, elevated post-RT Gal-9 in patient biopsies correlated with poor therapeutic outcomes. Notably, combining Gal-9 blockade with RT elicited potent abscopal responses in homologous two-tumor mouse models. Furthermore, anti-Gal-9 markedly enhanced radio-immunotherapy efficacy in the poorly immunogenic Lewis lung carcinoma. Immunologically, Gal-9 blockade synergized with RT to activate the myeloid and T cell compartments, enhancing dendritic cell accumulation in tdLNs and boosting CD8+ T cell infiltration in abscopal tumors. Depletion of CD8+ T cells/monocytes or blocking lymphocyte egress from lymph nodes abrogated the abscopal efficacy, underscoring their essential roles. Our findings establish RT-induced Gal-9 as a novel dual myeloid/T-cell immune checkpoint restricting abscopal responses. Gal-9 blockade represents a promising strategy to potentiate radiotherapy against metastatic disease, defining a therapeutic paradigm distinct from conventional checkpoint inhibitors.

Cysteine metabolism plays a crucial role in the growth and survival of non-small cell lung cancer (NSCLC), although the mechanisms governing its regulation are not fully understood. Here, we demonstrate that the RNA demethylase FTO is a therapeutic target that drives cysteine metabolism in NSCLC cells. Genetic or pharmacologic inhibition of FTO reduced cystine uptake and transsulfuration activity, leading to depleted intracellular glutathione, elevated reactive oxygen species (ROS), and ROS-mediated DNA damage and cell death. Mechanistically, FTO promotes the expression of the cystine uptake transporter SLC7A11 and the transsulfuration enzymes cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH) to promote NSCLC cystine uptake, transsulfuration activity, and survival. FTO inhibition increased lipid peroxidation, reduced tumor growth, and resulted in additive therapeutic benefit in combination with radiotherapy in multiple NSCLC xenograft models. Collectively, our study reveals a role for FTO in cysteine metabolism and highlights the therapeutic potential of targeting cancer epitranscriptomics and cysteine metabolism for NSCLC therapy.

Osimertinib, a third-generation EGFR tyrosine kinase inhibitor (TKI), has improved outcomes in non-small cell lung cancer (NSCLC) patients harboring the T790M mutation; however, emerging clinical evidence indicates a risk of cardiotoxicity. Here, we establish the first in vivo preclinical model of osimertinib-induced cardiotoxicity using transverse aortic constriction (TAC) in mice. Osimertinib treatment resulted in cardiac dysfunction, impaired hypertrophic remodeling, and increased markers of heart failure and fibrosis. Unbiased transcriptomic profiling revealed a myocardial stress response characterized by activation of p53-associated cell death pathways, mitochondrial dysfunction, and negative enrichment of histone acetyltransferase (HAT) complexes, indicating epigenetic repression. Mechanistically, osimertinib-treated hearts exhibited increased expression of multiple histone deacetylase (HDAC) isoforms, reduced acetylation of histones, and enhanced cardiomyocyte apoptosis via Bax/caspase-mediated pathways. There was a minimal, transient effect on inflammation, supporting a type I, cell-autonomous cardiotoxic mechanism. Consistent with this, in vitro and in vivo analyses demonstrated suppression of prosurvival ERK/AKT signaling, mitochondrial dysfunction, and activation of intrinsic apoptotic pathways. Given the central role of HDAC activation, we tested whether pharmacologic HDAC inhibition could mitigate osimertinib-induced cardiotoxicity. Treatment with the FDA-approved HDAC inhibitor vorinostat (SAHA) restored histone acetylation, attenuated p53 activation, reduced cardiomyocyte death, and rescued cardiac function in osimertinib-treated mice. Translational studies in human NSCLC-derived PC9 cells further demonstrated that SAHA enhances osimertinib antitumor efficacy while alleviating cardiotoxicity. Collectively, these findings define HDAC-dependent epigenetic repression as a key mechanism underlying osimertinib-induced cardiotoxicity and identify HDAC inhibition as a therapeutically actionable strategy to improve both cardiac safety and cancer treatment efficacy.

Loss-of-function STK11 mutations occur in 15% to 20% of lung adenocarcinomas (LUAD) and correlate with immunotherapy failure and worse survival. By integrating analysis of human tumor samples, a human LUAD cell line panel, and Cancer Cell Line Encyclopedia and The Cancer Genome Atlas datasets, we found that C3 production was increased in human STK11-mutant versus STK11-wildtype (WT) LUAD, modulated by LKB1 loss, whereas high C3 expression in STK11-mutant LUAD was associated with worse survival. STK11 knockout (STK11-KO) in syngeneic murine LUAD tumors resulted in increased neutrophil and reduced T-cell infiltration and anti-PD-1 resistance. STK11-KO tumor growth was similar in C3-/- versus WT mice. In contrast, C3 deletion in STK11-KO tumors resulted in dramatic inhibition of tumor growth and enhanced sensitivity to anti-PD-1 in immunocompetent mice but had no significant effect in CD8-depleted WT mice or nude mice, pointing to tumor-derived C3 promoting immune evasion. Mechanistically, STK11 loss drove tumor-derived C3 production and downstream CXCL2 and complement factor H production that promoted immune evasion and impaired anti-PD-1 efficacy. Our results show a C3-driven signaling axis for STK11-mutant LUAD promoting immune evasion and identify therapeutic targets to render these tumors sensitive to anti-PD-1.

The mechanisms by which mutations of splicing factor gene U2AF1 contribute to lung adenocarcinoma pathogenesis are not well understood. Here we used prime editing to modify the endogenous U2AF1 gene in lung adenocarcinoma cells and assessed the impact on alternative splicing. One specific KRAS mutation, G12S, led to skipping of KRAS exon 2 and generation of a nonfunctional KRAS transcript. However, expression of the U2AF1S34F mutant reverted this exon skipping and restored KRAS function, leading to enrichment of U2AF1S34F mutations in KRASG12S-mutant lung adenocarcinomas. Comprehensive analysis of splicing factor-oncogene mutation co-occurrence in cancer genomes also revealed significant coenrichment of KRASQ61R and U2AF1I24T mutations. Experimentally, KRASQ61R mutation led to KRAS exon 3 skipping, which in turn could be rescued by expression of U2AF1I24T. Our findings provide evidence that splicing factor mutations can rescue splicing defects caused by oncogenic mutations in a dynamic process of cascading selection.

The interplay between tumour cells and tumour-associated macrophages (TAMs) within the tumour microenvironment is crucial for the progression of non-small cell lung cancer (NSCLC). The underlying mechanisms involving RNA modification and exosomal communication remain incompletely understood. Multiplex immunofluorescence and flow cytometry were performed to evaluate M2 macrophage polarization. Exosomes were isolated by ultracentrifugation and validated by transmission electron microscopy, nanoparticle tracking analysis, and exosomal marker blots. To investigate the molecular mechanism, methylated RNA immunoprecipitation (MeRIP)-qPCR and dual-luciferase reporter assays were used to validate m6A modification sites on NFIC and miR-194-5p; RNA immunoprecipitation (RIP) confirmed the interaction between ZNF106 and interleukin-6 (IL-6) mRNA; chromatin immunoprecipitation (ChIP) was employed to detect STAT3 binding to the METTL3 promoter. The in vivo function of the identified feedback loop was assessed using an orthotopic xenograft mouse model of NSCLC. A negative feedback loop between METTL3 and NFIC was demonstrated in NSCLC cells. METTL3 suppressed miR-194-5p expression and its loading into exosomes through m6A methylation. NSCLC-derived exosomal miR-194-5p was internalized by macrophages and directly targeted ZNF106, thereby inhibiting M2 polarization. In macrophages, ZNF106 stabilized IL-6 mRNA and promoted exosomal IL-6 secretion, thereby activating the JAK2/STAT3 pathway and upregulating METTL3. This IL-6-driven METTL3 upregulation formed a positive feedback loop that sustains M2 polarization and tumour progression. In vivo disruption of this loop reduced tumour growth and metastasis. These findings establish a closed regulatory circuit initiated by an NFIC/METTL3 negative feedback loop. In this circuit, METTL3-mediated m6A modification of exosomal miR-194-5p in NSCLC cells derepresses ZNF106 expression in macrophages, leading to IL-6 production that activates the JAK2/STAT3 pathway and upregulates METTL3 in tumour cells, thereby perpetuating M2 polarization and malignant progression. This circuitry offers potential nodes for therapeutic intervention in NSCLC. NFIC/METTL3 negative feedback loop in NSCLC cells suppresses exosomal miR-194-5p via m6A methylation; reduced miR-194-5p deepresses ZNF106 in macrophages, promoting M2 polarization and IL-6 secretion; Macrophage-derived IL-6 activates JAK2/STAT3 in NSCLC cells to upregulate METTL3, forming a positive feedback loop.

BACH1 is a transcriptional regulator that modulates various cytoprotective pathways. Among these pathways BACH1 regulates cellular oxidative stress responses by suppressing the expression of cytoprotective genes. Dysregulated BACH1 activity has been implicated in a range of pathologies, including chronic inflammatory diseases, fibrosis, and cancer, making it a promising therapeutic target. However, BACH1 remains an underexploited drug target, with limited pharmacological inhibitors available. We have developed a novel luciferase-based reporter cell line enabling quantitative, high-throughput assessment of BACH1 inhibition. Using this platform, we rigorously screened two small-molecule libraries with 2046 compounds and identified four structurally distinct compounds that robustly inhibit BACH1 function. Notably, these compounds also activate transcription factor NRF2, suggesting the potential for a broader modulation of oxidative stress pathways. Importantly, we demonstrate that commonly used 2D migration assays may fail to detect phenotypes consistent with BACH1 inhibition, resulting in false negatives. In contrast, we establish that 3D invasion assays more robustly capture anti-invasive effects of BACH1 functional inhibition. Using this 3D system, we validate the identified compounds as potent suppressors of lung cancer cell invasion in vitro. This study delivers a novel screening platform for BACH1-targeted drug discovery, and challenges current in vitro standards by establishing 3D invasion assays as a more accurate functional readout for BACH1-targeting compounds. Additionally, it identifies new dual functional BACH1 inhibitors/NRF2 activators, offering novel chemical scaffolds for the development of anti-metastatic therapies and potentially treatments for diseases driven by oxidative stress and inflammation.

Platinum-based chemotherapy is the standard first-line treatment for advanced lung adenocarcinoma (LUAD). However, cisplatin resistance often leads to therapeutic failure, which subsequently results in cancer recurrence and metastasis. Here, we identify NSUN5 as a key driver of cisplatin resistance in response to oxidative stress. Cisplatin-induced reactive oxygen species (ROS) not only elevate NSUN5 expression through Nrf2-dependent transcriptional activation, but also enhance its methyltransferase activity by potentiating Cys359 for RNA capture. NSUN5 overexpression is clinically correlated with cisplatin resistance and poor prognosis in LUAD. Mechanistically, integrated RNA bisulfite sequencing (Bis-seq) and RNA sequencing (RNA-seq) analyses identified GLUT1 as a key target. NSUN5 catalyzes m5C methylation of GLUT1 mRNA through two functional sites, Cys359 for RNA binding and Cys308 for the release of m5C-modified RNA. YBX1 subsequently recognizes and stabilizes this m5C-modified transcript. Ultimately, NSUN5 enhances glucose uptake in a GLUT1-dependent manner, leading to increased glycolysis and thereby promoting homologous recombination (HR) repair. Functional studies confirmed that NSUN5 knockdown sensitized LUAD cells to cisplatin through increasing DNA damage, whereas its exogenous expression produced opposite effects. Crucially, silencing GLUT1 partially reversed NSUN5-mediated cisplatin resistance in vitro and in vivo. In summary, NSUN5 acts as a key translator that converts oxidative stress signals into enhanced HR repair by modulating the Warburg effect, thus highlighting its therapeutic potential for overcoming acquired chemoresistance in LUAD.

LKB1 mutations in lung cancer promote an immunosuppressive tumor microenvironment, but the underlying mechanisms remain unknown. Using genetically engineered mouse models and human tumor samples, we demonstrate that LKB1 loss leads to high expression of the cytokine leukemia-inhibitory factor (LIF), which through a cancer cell-autonomous autocrine loop, orchestrates the infiltration of immunosuppressive SiglecFHi neutrophils and Arg1+ interstitial macrophages. Genetic deletion of Lifr, the receptor for LIF, on Lkb1-mutant lung tumors revealed that autocrine LIF signaling induces tumor plasticity and the emergence of a Sox17+ dedifferentiated inflammatory cell state. Antibody-mediated LIF neutralization selectively eliminates the Sox17+ tumor cell state, reduces immunosuppressive myeloid cells, and enhances antitumor T-cell responses. Our study uncovers a novel LKB1-LIF axis driving immune evasion and identifies LIF as a potential therapeutic target in LKB1-mutant lung cancer. This work highlights the interplay between tumor genetics, cellular plasticity, and immune regulation in lung cancer progression. LKB1-mutant lung cancers express LIF, which induces an immunosuppressive Sox17+ tumor state. Anti-LIF therapy eliminates this state and restores antitumor immunity, revealing a novel vulnerability in this aggressive cancer subtype lacking effective targeted therapies.

The tumor suppressor p53 is pivotal in repressing tumorigenesis under physiological conditions. Paradoxically, we find that wild-type (WT) p53 plays an oncogenic role in relieving METTL5 depletion-caused cancer regression by sustaining mitochondrial respiration. The methyltransferase METTL5 is upregulated in non-small cell lung cancer (NSCLC) and associated with advanced tumor grade and poor prognosis. Depletion of METTL5 impairs NSCLC cell proliferation and migration in vitro and in vivo, with p53-null cells displaying enhanced sensitivity. While METTL5-depletion inhibits cytoplasmic translation in both p53-WT and p53-null cells, only cells lacking p53 exhibit severe tumor regression due to defective mitochondrial protein synthesis and consequent respiratory dysfunction. Mechanistically, p53 binds 5'UTR of TOMM40, the crucial gatekeeper of mitochondrial protein import, to enforce its exclusion from translation. METTL5 loss promotes p53 nuclear retention via inhibiting MDM2-mediated p53 ubiquitination, alleviating its translational suppression of TOMM40, and supporting oxidative phosphorylation. Remarkably, the combination targeting of p53 and METTL5 synergistically attenuates the proliferation and migration in p53-WT cancer cells. Our study elucidates the essential role of p53 in supporting tumor viability upon METTL5 deficiency by maintaining mitochondrial respiration. Meanwhile, it provides a molecular foundation for developing therapeutic strategies regarding cancers with WT p53.

Cardiovascular disease (CVD) remains the leading cause of mortality in women, yet sex-specific risk factors are usually not included in conventional predictive models. Specifically, heart failure (HF) in women may be influenced by sex-specific hormones and pathologies that need to be addressed to improve prevention and treatment. This expert consensus statement aims to provide a comprehensive roadmap for HF prevention and management across specific conditions affecting women during their life-course. Each section focuses on the impact of a specific female condition on CVD and how to prevent and manage HF in specific settings: 1. Pregnancy with a specific focus on how to deal with hypertensive disorders in the acute and chronic setting and how to prevent and treat Peripartum Cardiomyopathy (PPCM); 2. Gynecological conditions predisposing to HF, such as Polycystic Ovary Syndrome (PCOS), endometriosis, and the menopausal transition. Emphasis is placed on chronic inflammation, metabolic dysfunction, and the "window of opportunity" for Menopausal Hormone Therapy (MHT); 3. Cardio-Oncology: mitigating Cancer Therapy-Related Cardiac Dysfunction (CTRCD) in breast and gynecological cancers, focusing on female-specific cardiotoxicity profiles, the importance of subclinical detection of cardiac dysfunction and the implementation of cardioprotective strategies (ACE-inhibitors, Beta-blockers, SGLT2 inhibitors) during cardiotoxic treatments. Lifestyle interventions such as the DASH diet and exercise-based rehabilitation are highlighted as essential for maintaining cardiac reserve.

The rapid expansion of immunotherapy and targeted oncologic treatments leading to improved overall survival has led to a growing number of patients requiring dermatologic surgery (DS) for skin cancer while receiving systemic cancer therapy. However, perioperative management in this setting remains poorly defined. Dermatologic surgeons and oncologists must balance risks of impaired wound healing, bleeding, and infection against potential consequences of skin cancer progression versus treatment interruption, underscoring the need for multidisciplinary guidance. Here, we review perioperative management strategies at a tertiary cancer center and integrate FDA prescribing information, published data, and expert recommendations developed through multidisciplinary meetings between Mohs surgeons and medical oncologists. Our recommendations address key therapeutic classes including targeted therapies, cytotoxic chemotherapies, immunomodulators, proteosome inhibitors, and antibody drug conjugates, each of which presents distinct considerations. Across therapies, optimal management centers on individualized decision-making, preoperative laboratory assessment, careful timing of drug interruption for agents with known wound-healing or hematologic effects, and close postoperative monitoring. As systemic therapies continue to evolve, these expert-based recommendations aim to support safe, personalized perioperative care for the increasing population of oncology patients undergoing DS.

The 2026 European Association of Urology and American Society of Clinical Oncology (EAU-ASCO) guideline update reflects significant developments in the diagnosis and management of penile cancer. This review summarises the key changes and contrasts them with previous recommendations, with particular focus on staging, treatment, quality of life and emerging personalised approaches. The summary is based on a critical appraisal of the full 2026 guideline and its underpinning systematic reviews, with comparison to earlier versions. Recommendations were informed by structured literature assessment and expert panel consensus, incorporating evaluation of benefits and harms, evidence uncertainty and patient values. Major updates include refined pathological risk stratification, routine ultrasound (US)-guided nodal assessment, and broader guidance on organ-preserving surgery. There is support for selective genomic testing and clearer, restructured algorithms are introduced, including newly developed flow diagrams for nodal management, alongside an expanded evidence base for systemic therapy. Greater emphasis is placed on survivorship, centralisation of care and rationalisation of follow-up. However, many recommendations remain informed by retrospective data and expert consensus, reflecting the rarity of the disease and limited prospective evidence. The updated guideline promotes more nuanced selection of organ-preserving strategies, earlier detection of regional lymphatic disease, and holistic palliative care, while reinforcing the central role of shared decision-making. The new guidance for penile cancer aims to improve care, personalise treatment and better address quality of life, while acknowledging that further research is still needed.

Evidence on topical combined 5-fluorouracil/calcipotriene (5-FU/C) treatment of actinic keratoses (AKs), squamous cell carcinoma in situ (SCCIS), and superficial basal cell carcinoma is lacking. To develop an expert consensus regarding 5-FU/C treatment of AKs, SCCIS, and superficial basal cell carcinoma. Expert panelists were provided 5-FU/C literature review findings and survey results from International Immunosuppression and Transplant Skin Cancer Collaborative clinician members who prescribe 5-FU/C. Nine dermatologist/dermatologic surgeon International Immunosuppression and Transplant Skin Cancer Collaborative members with >5 years post-training experience or considerable clinical experience prescribing 5-FU/C participated in the panel. A 70% or higher concordance among respondents represented consensus. Key recommendations include: treating AKs with 5-FU/C twice daily for 4 to 5 days on the face/neck and 7 to 10 days on the scalp, trunk, and extremities; treating SCCIS with 5-FU/C twice daily for 7 to 10 days on the face/neck and 10 to 14 days on the scalp, trunk, and extremities; treating actinic cheilitis with 5-FU/C twice daily for 4 to 5 days; and no treatment modifications for patient immunosuppression. Small panel size and literature to date primarily including small, retrospective studies. Dermatologists may consider implementation of expert panel recommendations for treatment of AKs, SCCIS, and sBCC with 5-FU/C while awaiting larger, high-quality studies.

While dermatoscopy's added value to clinical imaging is known, the standalone value of dermatoscopy and clinical images remains unclear. To quantify the standalone and complementary value of dermatoscopy and clinical close-up imaging across lesion types and reader expertise. In an online reader study, 283 participants diagnosed 1567 paired images (clinical close-up/dermatoscopic) of skin cancers and mimics. The presentation order was randomized. Diagnostic accuracy, sensitivity, and specificity were compared. The effects of modality, reader expertise, and presentation order were analyzed using a generalized linear mixed model. Dermatoscopy alone showed higher sensitivity (85.0% vs 74.2%) but lower specificity (66.8% vs 71.9%) compared with clinical close-up images alone. It improved accuracy for melanoma and basal cell carcinoma, but not for nevi. Adding either modality increased the odds of a correct diagnosis. Dermatoscopy raised the odds by 52% (OR = 1.52; 95% CI: 1.36-1.70; P < .001), while clinical close-ups increased the odds by 40% (OR = 1.40; 95% CI: 1.20-1.61; P < .001). Image-based evaluation simulated teledermatology rather than face-to-face assessment; potential selection and verification bias cannot be excluded. Dermatoscopy alone yields higher sensitivity for malignant lesions but lower specificity for nevi than clinical close-up images. The latter provides complementary diagnostic cues, underscoring the value of integrating both modalities for optimal assessment.

Testosterone deficiency (TD) is a common condition affecting patients' health and quality of life. Clinical management has changed over the past decades, particularly regarding the indications and outcomes of testosterone therapy (TTh). Our aim was to provide an overview of the available evidence supporting the use of TTh in adult men with TD. A nonsystematic literature review was conducted to identify relevant studies on the diagnosis and treatment of TD. The review encompassed lifestyle and pharmacological approaches, summarizing recent advances and highlighting persisting gaps in clinical practice. Testosterone deficiency in adult men typically presents with mild, nonspecific symptoms often related to aging, including sexual dysfunction, fatigue, mood changes, and reduced muscle and bone mass. Diagnosis requires a thorough clinical assessment and confirmation of low serum total testosterone. TTh is the standard treatment for symptomatic men with TD and low testosterone levels. Evidence indicates that TTh improves sexual function, body composition, metabolic profile, and bone mineral density without increasing the risk of major cardiovascular events or prostate cancer. Current guidelines recommend maintaining testosterone within the midnormal range (450-600 ng/dl) and monitoring hematocrit, prostate-specific antigen, and metabolic parameters. Follow-up should occur at 3 mo and every 6-12 mo thereafter, with individualized adjustments based on clinical response and safety. TTh is a safe and effective treatment for TD. The selection of the most suitable product should consider patient needs and preferences, as well as the physician's perspective.

Keratinocyte carcinomas are the most common malignancy in the U.S. and may substantially affect quality of life (QoL). Few studies have assessed both general and skin cancer-specific QoL or examined how patient-level factors contribute to these outcomes. To assess general and skin cancer-specific QoL in individuals with keratinocyte carcinomas and identify demographic and clinical predictors of QoL impairment. This cross-sectional study included 1138 patients undergoing treatment for keratinocyte carcinomas at a single academic center. Participants completed surveys assessing demographics, disease history, health care utilization, and QoL using the Skin Cancer Index, short form-20, and a care burden score. Field cancerization classification categorized cumulative skin cancer history. Younger age, female sex, family history of skin cancer, higher field cancerization class, frequent dermatology visits, and chemoprevention use were significantly associated with poorer Skin Cancer Index and short form-20 scores. QoL appeared most impacted in the emotional and appearance domains of the Skin Cancer Index and the health perception domain of the short form-20. Single-center design, cross-sectional data, and self-reported measures. Identifying risk groups and the specific quality of life domains they value most may help clinicians deliver more personalized education and support strategies to improve patient-centered care.

Squamous cell carcinoma (SCC) is one of the most frequent solid tumor accounting for more than one million cancer deaths annually. Emerging evidence highlights the critical role of mechanical signaling in tumor progression, yet the mechanobiological interplay between extracellular matrix (ECM) stiffness and SCC pathogenesis remains poorly understood. This study aimed to investigate how matrix stiffness drives SCC progression via Piezo1-mediated mechanotransduction and its clinical implications. Atomic force microscopy quantified tissue stiffness in human cutaneous, oral, and lung SCC samples. Spatial transcriptomics and single-cell RNA sequencing (scRNA-seq) analyzed ECM- and mechanoreceptor-related gene expression. In vitro models using tunable-stiffness gels assessed SCC cell proliferation, invasion, and Piezo1/YAP activity. Xenograft models evaluated Piezo1's role in tumor growth and stiffness regulation. Clinical correlation studies analyzed Piezo1 expression and outcomes in a cutaneous SCC cohort (n = 53). SCC tissues exhibited elevated stiffness compared to adjacent normal tissues. Stiff matrices activated Piezo1, promoting proliferation and invasion in SCC cells via a non-canonical Hippo pathway involving RCC2. Piezo1 knockdown reduced tumor growth and stiffness in vivo. Mechanistically, Piezo1 activation induced TGFβ1 secretion, driving fibroblast-to-myofibroblast transition and collagen deposition, thereby reinforcing matrix stiffness. Clinically, high Piezo1 expression correlated with poor differentiation (p = 0.0034), recurrence (p = 0.047), and shorter disease-free survival (HR = 181.03, p = 0.01). This study identifies a self-reinforcing Piezo1-RCC2-YAP axis that translates mechanical cues into pro-tumorigenic signaling, fostering fibroblast-mediated ECM remodeling. Piezo1 emerges as a prognostic biomarker and therapeutic target to disrupt stiffness-driven SCC progression.

A 78-year-old man with hormone-sensitive metastatic prostate cancer underwent chest CT, which revealed a gradually enlarging pulmonary nodule in the left upper lobe, suggestive of lymphangitic metastasis. To further evaluate the lesion, both 18 F-FDG and 18 F-PSMA-1007 PET/CT were performed. The lesion showed intense FDG uptake but relatively low PSMA uptake, raising the suspicion of a primary lung malignancy. Histopathologic examination after wedge resection confirmed pulmonary tuberculosis.

Direct oral anticoagulants (DOACs) have transformed the prevention and treatment of thromboembolic disease, offering predictable pharmacokinetics, fewer interactions, and improved safety compared with vitamin K antagonists. Over the past decade, DOACs have become the preferred oral anticoagulants for most patients with atrial fibrillation and venous thromboembolism; yet, important gaps remain in their optimal implementation across diverse clinical scenarios. Despite strong evidence supporting their efficacy and safety, DOACs continue to be underutilized and sometimes inappropriately dosed, particularly among patients at elevated thrombotic risk and in populations historically underrepresented in randomized trials. This American College of Cardiology Scientific Statement provides a comprehensive, evidence-based overview of established and emerging indications for DOAC therapy in primary and secondary prevention of thrombotic events. Key domains include anticoagulation in atrial fibrillation, acute and extended treatment of venous thromboembolism, management in special populations (including patients with chronic kidney disease, liver dysfunction, cancer, obesity, frailty, prior bleeding, and valvular heart disease), periprocedural anticoagulation, and antithrombotic strategies following stroke, left atrial appendage closure, and catheter ablation. The document also addresses practical considerations such as drug selection, dosing strategies, duration of therapy, bleeding risk assessment, management of anticoagulant-related bleeding, drug-drug interactions, adherence, and cost-related barriers to care. This statement is intended to complement existing clinical practice guidelines by synthesizing evolving data, clarifying areas of uncertainty, and supporting individualized, patient-centered decision making. By addressing both evidence and implementation challenges, this document aims to improve the appropriate and equitable use of DOACs across the spectrum of thrombotic disease.

Frozen section (Frozen) analysis is essential for intraoperative margin assessment in skin cancer surgery, but assessment is limited by freezing artifacts that obscure cellular detail. Formalin-fixed paraffin-embedded (FFPE) histopathology, the gold standard, provides higher quality but requires >24 h for processing. Herein, we developed and validated generative AI models to translate Frozen images into AI-generated FFPE images (GenFFPE) using 2594 slides from 283 cases across five major skin cancer types. Four unpaired image-to-image models (CycleGAN, CUT, AIFFPE, and SANTA) were trained and compared using quantitative metrics and expert rankings; CUT demonstrated the best overall fidelity. External validation and a visual Turing test (accuracy 60.2%) confirmed image realism. Among 55 discrepant cases, GenFFPE-based reassessment increased diagnostic concordance by 53.3%. AI-based Frozen-to-FFPE translation is, thus, feasible and clinically meaningful, offering a potential tool to improve intraoperative diagnostic reliability and support decision-making for challenging tumor types such as extramammary Paget's disease.

Chemical labeling of nucleic acids is essential to pinpoint the structure, localization, and function of RNA and DNA. Yet, reversible sequence-independent chemistries that can label native RNA and DNA remain poorly developed. Here, we describe Reversible Uridine Nitrilium-mediated Addition (RUNA), a reversible covalent chemistry that selectively modifies uridine and thymidine residues via N3 deprotonation and reaction with a nitrilium ion intermediate generated from an aldehyde and an isonitrile. The reaction forms a stable N3 adduct that can be quantitatively reversed by hydrolysis. By using reagents that are either membrane permeable or impermeable, we demonstrate the localization and function of DNA on exosomes. Although exosomes harbor nucleic acids, whether the latter are encapsulated in the exosome lumen or are surface-adhered is unknown. RUNA revealed that exosomes display DNA on their outer surface. The abundance of such surface DNA increases upon DNA-damage accumulation in cancer cells that are treated with a PARP inhibitor. This surface DNA drives exosome uptake by M2-polarized macrophages through scavenger receptors and triggers a shift toward an M1-like proinflammatory state. The selective labeling of surface DNA revealed an unexpected mechanism by which exosomes engage innate immune cells. RUNA is a versatile tool to analyze the nucleic acid content and functionality of extracellular vesicles in health and disease.

The naked mole-rat (Heterocephalus glaber) is a long-lived mammal with resistance to cancer and hypoxia, suggesting the evolution of robust proteostasis networks. The ribosome, central for protein synthesis, is key to cellular stress responses and has an unusual feature: the 28S rRNA split; however, the details of its organization remain unknown. Here, we present high-resolution cryo-EM structures of the naked mole-rat 80S ribosome in four states of the elongation cycle. The structures reveal a conserved overall architecture and rRNA modification landscape compared to other mammals, and provide an atomic-level view of the distinct break in the 28S rRNA. This cleavage event, located in the D6 expansion segment, is structurally stabilized by a network of interactions with surrounding ribosomal proteins, maintaining the integrity of the large subunit. Our comparative analysis revealed that this compensatory network preserves a canonical architecture that is nearly indistinguishable from intact mouse and human ribosomes. These findings resolve the structural basis of this distinct cleavage, showing that it is a stable, integrated feature whose function is likely linked to more subtle regulatory mechanisms, rather than inducing major structural rearrangements.

The prognosis of patients with metastatic/relapsed/refractory Ewing sarcoma (ES) is dismal. Natural killer (NK) cells are highly cytotoxic to ES but limited by resistance within the ES tumor microenvironment (TME). Here we sought to overcome ES resistance to NK cells by a combinatorial immunotherapy approach simultaneously enabling NK tumor-specific-targeting via chimeric antigen receptor (CAR) against a novel ES target interleukin-1 receptor accessory protein (IL1RAP), circumventing transforming growth factor beta (TGFβ)-mediated NK immunosuppression by TGFβ1-imprinting, increasing NK cell antibody-dependent cellular cytotoxicity (ADCC) via an anti-GD2 antibody dinutuximab, and improving NK cell persistence and ADCC by an IL-15 agonist, NKTR-255. Peripheral blood mononuclear cells were expanded into NK and TGFβ1-imprinted-NK (imNK) cells using antigen-presenting feeder cells co-expressing IL-21 and 4-1 BBL. Anti-IL1RAP-CAR messenger RNA was electroporated into NK or imNK cells. In vitro cytotoxicity assays were performed to investigate the efficacy of anti-IL1RAP-CAR-NK/imNK cell alone or combined with NKTR-255 and/or dinutuximab against ES cells. Xenograft mouse models of ES were used to investigate the antitumor efficacy of the combinatorial CAR-NK/imNK cell therapy against ES in vivo. Single-cell RNA sequencing and mass cytometry analyses of cells from xenograft tumors were performed to identify mechanisms of response/resistance to this combinatorial immunotherapy. We found that anti-IL1RAP-CAR-NK cells significantly and specifically enhanced NK cytotoxicity in vitro and decreased tumor growth and lung metastasis in vivo against IL1RAP+ES. TGFβ1-imprinting significantly enhanced in vitro cytotoxicity and tumor infiltration of CAR-NK cells, leading to significantly reduced tumor growth and improved animal survival in the ES orthotopic mouse model. Compared with single agent or double combinations, the triple combination of imprinted-CAR-NK (CAR-imNK) cells and NKTR-255 with dinutuximab had superior antitumor efficacy against IL1RAP+GD2+ ES. Mechanistic studies on single cells from the xenograft tumors revealed increased apoptosis of ES cells, upregulated expression of ligands to NK inhibitory receptors on ES cells, and enhanced mouse macrophage migration in the ES TME in response to the CAR-imNK+NKTR-255+dinutuximab therapy. Our preclinical data demonstrate that combinatorial innate immunotherapy leveraging tumor-targeting TGFβ1-imprinted IL1RAP-CAR-NK cells combined with an IL-15 agonist and an anti-GD2 antibody is a promising novel therapeutic strategy for targeting metastatic/relapsed/refractory ES.

Amino acid storage proteins, such as serum albumin in mammals, are known to accumulate within tumors, but their precise contribution to tumor biology remains unclear. Using both cachectic and noncachectic tumor models in Drosophila, we observed that fat body-derived hexamerins accumulate in the tumors through a selective uptake process. Disabling this uptake led to a marked reduction in tumor growth, demonstrating that hexamerins are used as a nutrient source to support cancer progression. Hexamerin uptake also supports expression of the relaxin-like Drosophila insulin-like protein 8 (Dilp8) by the tumor, inhibiting ecdysone production and extending the growth period. This coupling between nutrient uptake by the tumor and inhibition of the developmental progression exerts a full diversion of the host resources. Functional parallels with mammalian albumins suggest evolutionarily conserved mechanisms with potential implications for cancer biology.

Extracellular ATP (eATP) accumulates in the tumor microenvironment (TME), where its metabolite adenosine is implicated in promoting immunosuppressive pathways. We found that immunotherapy further increased eATP concentrations in the TME. eATP directly signaled through the purinergic receptor P2RY2 in tumor cells to suppress T cell function, independently of its metabolites. Mechanistically, eATP-P2RY2 signaling drove expression of cyclooxygenase (COX)-1 and COX-2 and downstream accumulation of prostaglandin E2 (PGE₂). Genetic deletion or pharmacologic inhibition of P2RY2 reduced baseline intratumoral PGE₂ and prevented therapy-induced PGE₂ accumulation, changed the TME immune infiltrate, restored antitumor T cell responses, and overcame resistance to immune checkpoint blockade, CAR-T, and TCR-T therapies. A P2RY2 antagonist monoclonal antibody recapitulated the in vivo therapeutic benefit and enhanced human autologous tumor-infiltrating lymphocyte responses against matched tumor cells ex vivo. Thus, persistent eATP-P2RY2 signaling in the TME drives a PGE₂-mediated immunosuppressive circuit that operates at baseline and is further amplified during immunotherapy, driving adaptive resistance.

SUMMARYThe messenger RNA (mRNA)-lipid nanoparticle (LNP) vaccine platform is the newest tool in the vaccination arsenal for combating infectious diseases, rare genetic diseases, and cancer. Since 2020, major advances in preclinical and clinical studies have demonstrated the efficacy of mRNA-LNP vaccines against viral pathogens and in cancer. However, studies investigating the efficacy of mRNA-LNP vaccines against bacterial pathogens remain sparse due to substantial bacterial vaccine-specific challenges. Here, we highlight key challenges impeding the development of mRNA-LNP vaccines against human bacterial pathogens, the advantages that can be gained by utilizing the mRNA-LNP platform for bacterial vaccines, and progress toward the development of mRNA-LNP vaccines against bacterial infections.

Sex-based differences in cancer incidence are incompletely understood, but potential roles for the immune system are beginning to emerge. CD4+ T cells play a central role in coordinating antitumor immunity. In addition to cytokine production, CD40L expression on CD4+ T cells provides necessary helper signaling to dendritic cells (DC) that is required for the priming of cytotoxic tumor-specific CD8+ T cells. Despite these critical functions, the impact of biological sex on the CD4+ T-cell response to cancer remains unknown. In this study, we demonstrate that impaired immune-mediated tumor control in male mice compared with female mice is driven by disparate CD4+ T-cell responses in a mouse model of bladder cancer. We found that CD40L expression was reduced on CD4+ T cells isolated from males via a mechanism predominantly driven by cell-intrinsic androgen receptor signaling, resulting in decreased DC licensing through CD40 within tumor-draining lymph nodes. These deficits resulted in decreased helper CD4+ T-cell frequencies and impaired CD8+ T-cell function within the male tumor microenvironment, which could be rescued by targeting the CD40L-CD40 axis. Our findings identify a novel mechanism of CD4+ T cell-based sex differences in the immune response to cancer that impairs tumor control. See related Spotlight, p. 1052.

Evaluating the impact of bias in observational epidemiology studies is critical to systematic reviews in public health (as is the case with any study type). However, researchers have expressed concern that useful epidemiological data may not receive full consideration in some reviews due to overemphasizing potential for bias without critical assessment of its impact. This concern should be balanced against the need to appropriately assess the degree of bias in observational studies to accurately characterize their validity and reach appropriate conclusions. In this paper, we describe how examining bias across studies may alleviate or solidify concerns identified in individual studies. We describe how to assess the likelihood that specific sources of bias explain and/or meaningfully distort an observed association. We followed the methods for development of Grading of Recommendations, Assessment, Development, and Evaluations (GRADE) workgroup guidance. Various sensitivity analyses are proposed to probe risk of bias in the body of evidence. Two illustrative case studies were conducted: the associations between trichloroethylene (TCE) exposure and kidney cancer occurrence and between two individual per-and polyfluoroalkyl substances (PFAS) and changes of alanine aminotransferase (ALT) levels. Sensitivity analyses can inform the potential for bias to influence the results of a body of evidence, including consideration of direction of bias, subsets/stratification, counterbalancing biases, use of positive/negative controls, and others. To incorporate these into a synthesis, consideration is needed throughout the planning and execution process: (1) Planning the systematic review; (2) Assessing risk of bias in individual studies; (3) Performing sensitivity analyses, including exploration of heterogeneity and bias impact; and (4) Determining final rating. These nuanced concepts and approaches for more accurately characterizing the magnitude and impact of bias in a body of evidence can be applied within GRADE and more broadly to systematic reviews outside that framework.

NRF2 (Nuclear Factor Erythroid 2-Related Factor 2) has traditionally been viewed as a master regulator of the antioxidant response. However, emerging evidence redefines NRF2 as a pleiotropic and evolutionarily conserved systems-level integrator of redox, metabolic, proteostatic, and inflammatory homeostasis, i.e., an evolutionarily conserved guardian of health. From its origins as an adaptive response to rising atmospheric oxygen in early metazoans to its role in orchestrating complex cytoprotective networks in humans, NRF2 exemplifies the biological transition from stress resistance to adaptive resilience. Understanding its regulation across species, tissues, life stages and gender offers novel perspectives for combating age-related cellular dysfunction and reframing the dynamic process of redox regulation as a component of adaptive homeostasis in a precision medicine perspective. This review summarizes and integrates essential concepts currently being developed regarding the evolutionary roots of NRF2, its molecular and regulatory complexity, its dualistic role in health and disease, and its potential as a biomarker of resilience and therapeutic target. We highlight NRF2's function as a rheostat rather than a binary stress switch, including the role of hormetic activation through lifestyle and environmental stimuli. By maintaining redox homeostasis and dynamic adaptive stress responses, NRF2 bridges molecular defense mechanisms with strategies for healthspan extension and preventive medicine from very early life stages until senescence.

Transforming growth factor β (TGF-β) signaling is a highly pleiotropic pathway with an important role in development, homeostasis, and cancer. Chromatin regulators contribute to the regulation of TGF-β-responsive transcription. The requirement of subunits of the MLL3/MLL4 histone methyltransferase complexes for TGF-β responses has been reported. However, their exact roles are not fully understood. To investigate the functions of these complexes, we employed CRISPR/Cas9 genome editing to inactivate the KMT2C/MLL3 or KMT2D/MLL4 genes in human diploid epithelial cells. Time-course RNA-seq experiments revealed the requirement of MLL4 but not of MLL3 for TGF-β transcriptional responses. CUT&RUN experiments showed that MLL4 binding increases after TGF-β treatment and is especially enriched at AP-1 transcription factor binding sites. Interestingly, TGF-β-induced chromatin binding of MLL4 correlates with increases in H3K27ac but not in H3K4me1 modifications. Furthermore, TGF-β treatment sets off SMAD2-induced JUNB expression, which forms a feed-forward loop with MLL4. By inhibiting the activities of AP-1, the BAF chromatin remodeler, or the CBP/p300 histone acetyltransferase, we found that AP-1 binding and these chromatin regulators are all necessary for TGF-β induction of MLL4 binding and transcriptional activation of its genomic targets. Taken together, our study reveals distinctive roles for the MLL3 and MLL4 paralogs in the transcriptional response to TGF-β. In contrast to MLL3, MLL4 forms a feed-forward loop of JUNB, the BAF complex, and CBP/p300 to sustain transcription activation by TGF-β.

Cholangiocytes are specialized epithelial cells that line the intrahepatic and extrahepatic biliary tree and play a critical role in bile modification, liver homeostasis, and response to injury. Cholangiocytes exhibit notable heterogeneity and plasticity, and their dysfunction is central to a spectrum of diseases targeting the bile ducts, collectively called cholangiopathies. These disorders include genetic, infectious, immune-mediated, and malignant diseases, with primary sclerosing cholangitis (PSC) representing one of the most complex and enigmatic of these disorders. PSC is a progressive, fibro-inflammatory disease of the bile ducts that is closely linked to inflammatory bowel disease, carries a heightened risk of cancer, and lacks any approved therapies. This review explores the biology of cholangiocytes, including their development, functional plasticity, and roles in secretion, absorption, and cellular signaling. We provide a detailed examination of cholangiopathies, particularly PSC, a complex cholangiopathy characterized by a paradoxical state of cholangiocyte senescence and hyperproliferation. We describe how immune cell dysfunction, the gut microbiome, genetic predispositions, and environmental factors converge to mediate PSC pathogenesis. We revisit the foundational technologies that empowered early discoveries and shaped the field as we know it today. We also explore how newer techniques such as organoid cultures, single-cell transcriptomics, epigenomics, and spatialomics have transformed our modern understanding of biliary pathophysiology. Finally, we provide an overview of existing rodent models of cholangiopathies and discuss their relevance to human disease. PSC remains therapeutically unaddressed, and thus ongoing multidisciplinary efforts are essential to developing targeted interventions. This review serves as a comprehensive resource for researchers and clinicians navigating the rapidly evolving landscape of cholangiocyte-centered liver disease research.

The identification of genetic risk factors could facilitate our understanding of the cause and pathogenesis of rheumatic diseases. Maps of susceptibility loci have been established for most complex diseases, including all major rheumatic diseases, but definitive causal variants have been identified only very infrequently. Here, we highlight the need to both position and confirm the role of causative polymorphisms in humans using experimental animals. The approach is best exemplified by the positional cloning of a causative single-nucleotide polymorphism (SNP) (rs201802280), which results in the substitution of arginine with histidine at position 90 in the NCF1 gene (neutrophil cytosolic factor 1, a component of the phagocyte NOX2 (Nicotinamide adenine dinucleotide phosphate oxidase 2) complex. This variant was independently identified and validated using 2 distinct approaches: by positioning the gene in animal models, followed by exon sequencing and validation of the identified SNPs, and by targeted resequencing and linkage disequilibrium mapping in multiethnic case-control studies. This causal SNP, located in a region with copy number variation, has emerged as a major susceptibility variant for multiple rheumatic diseases, including systemic lupus erythematosus, rheumatoid arthritis, primary Sjögren's disease, and systemic sclerosis. It represents the first major causal polymorphism conclusively linked to both increased susceptibility and disease severity across multiple rheumatic conditions, in both patients and animal models. The functional impact of the NCF1 variant has been explored mechanistically, and a viral infection has been identified as an interacting environmental factor contributing to lupus disease in animal studies. Importantly, the significance of the NCF1 polymorphism extends beyond rheumatic diseases-it has also been implicated in cancer, cardiovascular syndromes, infections, and physiological traits.

Gain-of-function mutations of isocitrate dehydrogenase 1 (IDH1) lead to oncometabolite (R)-2-hydroxyglutarate production, contributing to the tumorigenesis of multiple human cancers. While fatty acid biosynthesis is critical for IDH1-mutant tumor growth, the underlying mechanisms remain unclear. Here, leveraging chemical probes and chemoproteomic profiling, we identified that oncogenic IDH1-R132H is uniquely autopalmitoylated at C269, which is not observed in wild-type IDH1. This modification responds to fatty acids and regulates R132H enzymatic activity by enhancing substrate and cofactor binding, as well as dimerization. Loss of C269 palmitoylation reverses IDH1-R132H-induced metabolic reprogramming and hypermethylation phenotypes and impairs cell transformation. Interestingly, C269 autopalmitoylation occurs within a hydrophobic pocket, targeted by a clinical IDH1-mutant inhibitor (LY3410738). Our study reveals that autopalmitoylation, conferred by the IDH1R132H mutation, links fatty acid metabolism to the regulation of IDH1 mutant activity and represents a druggable vulnerability in IDH1-mutant cancers.

Keratinocyte carcinoma, including basal cell and squamous cell carcinoma, is the most prevalent form of cancer within the United States, with its incidence rapidly rising annually among patients over 65 years of age and those who are immunocompromised. Over the last decade, there have been multiple advancements made in the development of noninvasive diagnostic methods for identification of keratinocyte carcinoma, which have the potential to reduce the number of unnecessary skin biopsies. Part 1 of this CME will outline current and emerging noninvasive modalities for the diagnosis of keratinocyte carcinoma, including dermoscopy, reflectance confocal microscopy, optical coherence tomography, electrical impedance spectroscopy, tape stripping to isolate genetic material or proteins from the stratum corneum, and artificial intelligence. These methods are particularly helpful in patients with comorbidities or contraindications for skin biopsies, within cosmetically sensitive regions, and patients suffering from biopsy fatigue and/or fear.

Molecular motors, known for their ability to undergo controlled unidirectional rotation under external stimuli, have gained growing interest for their potential applications in biological systems and smart materials. Meanwhile, glycosylated macromolecules are utilized in lectin-binding, making them potential candidates to combat pathogenesis, cancer, and biofilms. However, glycopolymers are limited by their current inability to adapt in response to external stimuli, restricting their potential for future precision therapies. By integrating multivalent glycopolymeric structures with first-generation molecular rotary motors, it becomes possible to control carbohydrate-lectin interactions using intrinsic motor functions. Herein, we present photoadaptive glycopolymer-based first-generation molecular motors functionalized with immunologically important monosaccharides, β-d-glucopyranoside and β-d-mannopyranoside. The lectin-binding properties were subsequently investigated using surface plasmon resonance with DC-SIGN and Langerin, key players within the human innate immune system. Additionally, a competition assay with the envelope glycoprotein of HIV, namely gp120, was conducted to ascertain the inhibitory potential of these photoresponsive glycopolymer-based molecular motors. It was established that the light-controlled conformation of the molecular motor impacted both the overall binding affinity and half-maximal inhibitory concentration values. These findings create opportunities to control the binding affinity of glycopolymers and emphasize the potential of light-driven glycopolymers to modulate key interactions in cell-specific targeted delivery of therapeutics.

Testicular cancer is the most common solid tumor in young men aged 15-44 years, with seminoma being a major subtype. To elucidate its poorly understood molecular mechanisms, we performed integrated spatial transcriptomics (ST) and spatial metabolomics (SM) on fresh-frozen seminoma tissues from four patients. ST identified 16 distinct cell clusters and spatially resolved transcriptional programs in tumor, invasive, tubular, and stromal regions, revealing tumor-enriched pathways like glycolysis, oxidative phosphorylation, and ferroptosis. Pseudotime analysis suggested a branched trajectory of malignant progression. SM showed pronounced dysregulation of glutathione metabolism in tumor areas. Integrated analysis pinpointed glutathione metabolism and ferroptosis as convergent regulatory pathways in seminoma. Functional validation in TCam-2 cells and a xenograft mice demonstrated that pharmacologic inhibition of glutathione synthesis (using buthionine sulfoximine, BSO) or genetic knockdown of xCT induced ferroptosis and suppressed tumor cell growth both in vitro and in vivo. This study establishes glutathione metabolic reprogramming as a key adaptive mechanism in seminoma that maintains redox homeostasis and protects against ferroptosis, suggesting that targeting this pathway or promoting ferroptosis represents a novel therapeutic strategy.

In response to stress, cells exploit plasticity to self-activate and form protrusions that explore the environment and guide migration. While protrusion initiation is well characterized, their maturation and functional composition remain poorly understood. Here, using a new pooled genetic approach (ReGenT-seq) to interrogate chromatin factors controlling epidermal progenitor activation, we unexpectedly identified Cbx3 as a critical determinant of protrusion formation. Beyond its nuclear activities, we revealed a cytoplasmic moonlighting function of CBX3 that governs the subcellular localization of specific mRNAs within polarizing protrusions of migrating epidermal progenitor cells, as well as in invadopodia of squamous carcinoma cells. CBX3-dependent mRNA transport promotes the localized establishment of multiple organelles, including the endoplasmic reticulum and lysosomes, within maturing protrusions, and regulates focal adhesion dynamics. Together, our findings uncover a multilayered gene regulatory mechanism controlling cell motility through protrusionogenesis and identify a chromatin-to-cell-protrusion mRNA transport pathway that represents a potential therapeutic target for enhancing wound healing and limiting cancer invasion.

Radiotherapy for breast cancer exposes the heart to incidental radiation, and historical data have shown a dose-dependent increase in associated cardiac events. Although whole-heart dose metrics are commonly used for risk assessment, emerging evidence suggests that dose to the left anterior descending coronary artery (LAD) may better capture risk. To compare the ability of heart and LAD radiation dose metrics to predict cardiac risk after breast radiotherapy. This was a cross-sectional study of patients with breast cancer who were treated with either 3-dimensional conformal or intensity-modulated radiotherapy from 2008 to 2018 at a tertiary care center in Canada. The primary analysis included patients with left-sided breast cancer. Cardiac events were assessed using longitudinal follow-up data. Dosimetry was derived from computed tomography plans using automated segmentation and converted to equivalent dose in 2-Gy fractions (EQD2). Data were analyzed from September 2024 to April 2026. Adverse cardiac events defined as myocardial infarction or hospital admission or emergency department visit for unstable angina (ie, acute coronary syndrome), arrhythmia, heart failure, pericarditis, or myocarditis. The incidence of coronary angiography and coronary revascularization was also captured as coronary artery disease (CAD). Discrimination for dose metrics was performed with receiver operator characteristic curves and competing-risks regression (Fine and Gray), adjusted for cardiovascular risk factors. The analysis included 4908 patients with breast cancer of whom 2223 had left-sided breast cancer. During a median (IQR) follow-up period of 10.8 (8.4-13.1) years, cumulative incidence of cardiac event or CAD was 5.0 (95% CI, 4.1-6.0) at 10 years. A data-driven cut point analysis identified 12 Gy EQD2 as the maximum LAD dose that best stratified risk. Among patients with left-sided breast cancer, maximum LAD dose (concordance [C] index, 0.58; 95% CI, 0.52-0.64) discriminated better than mean heart dose (C index, 0.53; 95% CI, 0.47-0.60). In multivariable analysis, maximum LAD of 12 Gy or greater was independently associated with higher cardiac risk (subdistribution hazard ratio = 1.81; 95% CI, 1.04-3.16; P = .04), whereas mean heart dose of 2 Gy or greater was not associated (P = .99). For clinical context, the 12-Gy EQD2 for maximum LAD dose corresponds to a physical dose of approximately 10.5 Gy for 42.5 Gy in 16 fractions and 7 Gy for 26 Gy in 5 fractions. In this cross-sectional study of heart-sparing breast radiotherapy, LAD dose was associated with cardiac events, whereas whole-heart metrics was not. These findings support LAD-based planning and respiratory motion management to reduce long-term cardiovascular risk in patients with breast cancer undergoing radiotherapy.

In the US, breast cancer mortality is lower among Asian American females diagnosed with breast cancer compared with other racial and ethnic groups, although reasons for this difference are not well understood. To examine contributions of clinical, lifestyle, and sociodemographic factors to mortality differences among females identifying as Asian races and ethnicities compared with non-Latina White females and whether associations vary by nativity. This cohort study analyzed time-to-event survival data from Asian and White females with breast cancer from 4 epidemiologic studies. Cohorts were recruited from California and Hawaii, with information from questionnaires and cancer registries. Participants were females with first primary invasive breast cancer diagnosed from 1992 to 2019 who self-identified as Asian and females who self-identified as White as a comparison group. Analyses were conducted from February 2024 to February 2026. Self-reported race and ethnicity, nativity, clinical factors (eg, stage and tumor grade), marital status, educational level, reproductive factors, insurance status, lifestyle factors (eg, smoking and alcohol use), diabetes, and neighborhood socioeconomic indicators. The outcomes of interest were all-cause and breast cancer-specific mortality. Cox proportional hazards models were used to estimate hazard ratios (HRs) comparing each Asian group with White females, and changes in these estimates with adjustment for various prognostic factors. A total of 8994 females (mean [SD] age at diagnosis, 59 [13] years), including 3973 Asian females and 5021 White females, were analyzed. There were 2637 total deaths and 1140 deaths attributed to breast cancer (mean follow-up time 12.6 years; range, 1 month to 28 years). Most Asian groups had a higher proportion diagnosed before age 50 years compared with White females: from 34% among Filipinas to 73% among another Asian group vs 15% among White females. Nearly all Asian ethnic groups had lower proportions (45%-67%) of localized stage disease relative to White females (69%). Relative to White females, risk of all-cause mortality in fully adjusted models was lower for Chinese females (HR, 0.77; 95% CI, 0.62-0.96), Filipina females (HR, 0.81; 95% CI, 0.67-0.99), Japanese females (HR, 0.71; 95% CI, 0.54-0.93), and Asian females identifying as multiple races or ethnicities, not Native Hawaiian or Pacific Islander (HR, 0.67; 95% CI, 0.51-0.89). Lower adjusted risk of all-cause mortality was observed for Chinese (HR, 0.73; 95% CI, 0.57-0.94) and Filipina (HR, 0.79; 95% CI, 0.63-0.94) females born outside the US and Japanese females (HR, 0.63; 95% CI, 0.45-0.90) and Asian females with multiple races or ethnicities, not Native Hawaiian or Pacific Islander (HR, 0.59; 95% CI, 0.39-0.88) born in the US compared with White females. In this pooled cohort study of females with breast cancer, several Asian groups had lower risk of all-cause mortality, even after accounting for clinical, sociodemographic, reproductive, lifestyle, and neighborhood factors. Identification of the specific resiliency factors will provide insights into mechanisms that may improve survival after breast cancer.

What is the effect of group exercise-based telerehabilitation compared with a single session of in-person assessment and advice on health-related quality of life in cancer survivors? What are the effects on activity, function, safety and cost-effectiveness? An assessor-blinded, pragmatic randomised controlled trial with embedded cost analysis, concealed allocation and intention-to-treat analysis. Adult cancer survivors with any cancer diagnosis who were receiving treatment or within 12 months of treatment completion. The experimental group received an 8-week, twice-weekly, physiotherapist-led exercise group via videoconferencing, supplemented with support resources and a single in-person session of assessment and advice (TeleCaRe). The control group received a single in-person session of exercise assessment and advice. Assessments were completed at baseline, after the intervention (week 9) and at follow-up (week 26). The primary outcome was health-related quality of life, measured using the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire Core 30, at week 9. Secondary measures were walking capacity (6-minute walk test), physical activity (accelerometer), self-efficacy (Health Action Process Approach Questionnaire), adverse events, and health service and cost data. In total, 117 participants were recruited. Their mean age was 59 years, 82 (70%) were female, and 47 (40%) had breast cancer. Participants attended an average of seven out of 16 sessions (SD 6). There were no major adverse events. Intention-to-treat analysis found that TeleCaRe was not superior for improving quality of life (MD -5.3 units, 95% CI -13.3 to 2.6) or any secondary outcomes. TeleCaRe cost AU$363 per participant. Group exercise-based cancer telerehabilitation was safe but attendance was poor. The addition of telerehabilitation to assessment and advice was not superior to a single in-person physiotherapy session alone for improving quality of life. ACTRN12621001417875.

Ultrasound is widely used in breast cancer diagnosis due to its cost-effectiveness, non-invasiveness, and radiation-free properties. Computer-aided diagnostic systems based on dynamic breast ultrasound videos have advanced rapidly. However, existing methods are often affected by redundant information, make limited use of clinical prior knowledge, and provide insufficiently interpretable diagnostic results. To address these limitations, a clinical knowledge constrained multi-task learning framework (CKC-Framework), inspired by radiologists' workflow, is proposed for breast cancer diagnosis in ultrasound videos. The framework employs a DINO-based object detection algorithm to select keyframes and tumor-containing segments from variable-length videos, filtering out disruptive background frames. A diagnostic attribute constraint module is designed to enhance intra-frame feature interpretability and provide radiologists with keyframe diagnostic attributes. In addition, a clinical prior constraint module is introduced to guide temporal weighting and improve cross-frame semantic consistency. Finally, a keyframe-guided spatiotemporal Mamba is proposed to integrate spatial features across frames in long sequences via spatiotemporal scanning. For evaluation, we constructed Breast-USV, a large-scale breast ultrasound video dataset, and used the public BUSV dataset for benchmarking. Experimental results demonstrate that the framework achieves an AUC of 94.59% and an accuracy of 91.43% on Breast-USV, and an AUC of 90.38% and an accuracy of 87.39% on BUSV, achieving superior performance compared with existing methods, with additional cross-dataset evaluation from Breast-USV to BUSV to assess out-of-domain generalization. Moreover, by providing diagnostic attributes and lesion localization, the framework may support radiologists' decision-making and facilitate more efficient post-acquisition review.

The May 1 approval of vepdegestrant, the first PROTAC drug to earn regulatory clearance, establishes targeted protein degradation as a clinically validated therapeutic modality. For patients with ESR1-mutant advanced breast cancer, it offers a new therapeutic option after standard hormone-based regimens have failed. And for the broader field, it is proof of concept for a pipeline of protein-destroying drugs that now numbers in the dozens.

To compare HER2-targeted with ¹⁸F-FDG PET/CT for detecting axillary lymph node metastasis in patients with newly diagnosed HER2-positive or HER2-low breast cancer. In this prospective study, HER2-expressed breast cancer participants underwent both 18F-FDG and Al18F-NOTA-HER2-BCH PET/CT within 1 week. On the basis of IHC and/or FISH, participants were categorized as HER2-positive (n=25) or HER2-low (n=26). Histopathology or imaging follow-up served as reference standards. Statistical analyses utilized the paired McNemar test for diagnostic performance, and the Wilcoxon signed-rank and Mann-Whitney U tests for paired and unpaired comparisons of uptake parameters, respectively. Between August 2024 and October 2025, 51 participants with breast cancer were enrolled. Among 40 participants with lymph node metastasis, Al18F-NOTA-HER2-BCH detected metastatic lymph nodes exclusively in one HER2-positive participant and more lesions than 18F-FDG in 11 cases. On a per-lesion analysis, Al18F-NOTA-HER2-BCH demonstrated significantly higher sensitivity, accuracy, and negative predictive value than 18F-FDG for detecting metastatic axillary lymph nodes in the HER2-positive cohort (94%, 94%, 93% vs. 79%, 85%, 84%, all P<0.001). This superiority was particularly pronounced for small lymph nodes (<5 mm), with a detection rate of 90% versus 62% for 18F-FDG (P<0.001). Al18F-NOTA-HER2-BCH PET/CT directly influenced nodal staging and subsequent treatment decisions in 4 of 25 (16.0%) HER2-positive participants. In the HER2-low cohort, however, the diagnostic performance of the 2 tracers did not differ significantly, although 18F-FDG PET/CT led to clinically relevant nodal upstaging in one case. Al18F-NOTA-HER2-BCH retains developmental potential for detecting axillary lymph node metastasis in HER2-positive breast cancer, complementing 18F-FDG.

Despite the widespread adoption of chemoimmunotherapy in triple-negative breast cancer (TNBC), the mechanisms by which cytotoxic chemotherapy engages antitumor immunity remain poorly defined. Identifying tumor-intrinsic immunogenic programs that predict and enhance responsiveness to immune checkpoint blockade (ICB) is therefore of critical clinical importance. Transcriptomic signatures of TREX1 deficiency were generated from CRISPR-engineered TNBC models and applied to multiple independent TNBC cohorts treated with chemoimmunotherapy. Cytosolic single-stranded DNA (ssDNA) accumulation was quantified using a flow cytometry-based assay to functionally screen chemotherapeutic agents. Immune activation and therapeutic efficacy were evaluated using in vitro assays, syngeneic mouse tumor models, flow cytometry, and single-cell RNA sequencing. Statistical analyses included two-sided t tests or Wilcoxon tests, analysis of variance where appropriate, and receiver operating characteristic analyses. An in vivo TREX1-deficiency transcriptional signature, reflecting ssDNA-driven immune programs rather than TREX1 expression alone, robustly predicted clinical response to chemoimmunotherapy across independent TNBC cohorts. Functional screening identified LP-184, an acylfulvene-derived alkylating agent in clinical development, as a potent inducer of cytosolic ssDNA and type I interferon signaling. LP-184 enhanced antigen presentation, reduced immunosuppressive M2-like macrophages, promoted CD8+ T-cell priming, and synergized with anti-programmed cell death protein 1 therapy in vivo. Exogenous ssDNA recapitulated key immunostimulatory effects of LP-184, supporting ssDNA accumulation as a central mechanistic mediator. These findings establish cytosolic ssDNA-driven immune programs as a mechanistic link between DNA damage and antitumor immunity, providing both a predictive biomarker and a therapeutic axis for chemoimmunotherapy. Pharmacologic induction of ssDNA represents a rational strategy to enhance ICB efficacy and expand immunotherapy responsiveness in TNBC.

The objective of this study was to define the relationships between the circulating proteome and metabolome with cardiac structure and function in patients with breast cancer receiving cardiotoxic therapies. Proteomics and metabolomics profiling was performed in a longitudinal, prospective cohort study of breast cancer patients receiving anthracyclines and/or trastuzumab, using the Olink Explore 3072 platform and rapid liquid chromatography-mass spectrometry, respectively. Multivariable linear mixed-effect models evaluated the contemporaneous (same visit) and lagged (subsequent visit) associations between repeated measures of individual proteins or metabolites with quantitative echocardiographic measures of cardiac structure [left ventricular (LV) mass and left atrial volume index] and function [LV ejection fraction (LVEF), longitudinal and circumferential strain, E/e', and ventricular-arterial coupling]. Cox regression and pathway enrichment analyses were conducted for biomarkers demonstrating significant associations with cardiac function. Across 547 breast cancer participants (median age 50 years), 203 unique proteins and 16 unique metabolites were significantly associated with measures of cardiac structure and function in contemporaneous and lagged analyses. Notably, cathepsin C was associated with LVEF [false discovery rate (FDR), P = .017], longitudinal strain (FDR, P = .046), left atrial volume index (FDR, P = .035), and incident cardiac dysfunction, defined by an LVEF decline ≥10% to <50% (hazard ratio .61, 95% confidence interval .41, .90). The 147 proteins associated with cardiac function were enriched in biological processes reflective of protein deubiquitination, protein modification by small protein removal, macromolecule catabolic processes, and global metabolic pathways. Individual metabolites significantly associated with cardiac function (LVEF, longitudinal strain) included n-acetylglutamine, aspartic acid, acetylasparagine, alanyl-alanine, and prolyl-glycine (FDR, P-value < .001), and belonged to amino acids and derivatives and peptides. These findings provide translational insights into cancer therapy-related cardiac dysfunction and remodelling and identify potential new biomarkers of cardiotoxicity. There is an important need for validation of these findings and a deeper understanding of the biology of these biomarkers.

Activating the stimulator of interferon genes (STING) pathway is a potent strategy for promoting tumor vascular normalization and enhancing cancer immunotherapy. However, the poor metabolic stability of classical STING agonists limits their clinical translation. Herein, we engineered a self-assembled Au-Se nanoplatform (AAC) for the co-delivery of the STING agonist cyclic 2',3'-GMP-AMP (cGAMP) and the hypoxia-inducible factor-1α (HIF-1α) inhibitor acriflavine (Acf). Upon cellular internalization, the weakly acidic tumor microenvironment (TME) triggers the synchronized release of cGAMP and Acf agents. Next, cGAMP activates the STING pathway to induce type I interferon (IFN-I) production, while Acf disrupts HIF-1α/β dimerization. The activation of STING signal pathway and the inhibition of HIF-1α/β expression synergistically suppress the expression of VEGFA and remodels the TME into an immune-permissive milieu, which promotes vascular normalization and elicits tumor immune response. Combined with the photothermal therapy (PTT) of Au nanorods, AAC induces robust immunogenic cell death (ICD), facilitates dendritic cell (DC) maturation, and enhances cytotoxic T lymphocytes (CTLs) infiltration. In murine breast cancer models, AAC effectively suppressed both primary and distant tumors, highlighting its potential as a versatile nanotherapeutic platform for synergistic photothermal enhanced immunotherapy.

Women who have full-term pregnancies have a reduced risk of breast cancer, although the underlying mechanisms are not well understood. Here we show that tissue-resident memory-like T (TRM-like) cells are enriched in the breasts of parous women and mice compared to nulliparous individuals. These cells develop during mid-gestation, persist after lactation and are closely associated with mammary epithelial cells, suggesting dependence on epithelial-derived signals. Impaired alveolar differentiation or a deficiency in epithelial cell-derived cytokines interleukin (IL)-15 and transforming growth factor (TGF)β diminished the expansion of mammary TRM-like cells. Single-cell transcriptomics revealed the expanded TRM-like cells were heterogeneous, comprising CD8αβ+ and CD8αα+ subsets. Lineage tracing showed that these TRM-like cells acquire effector functions and contribute to tumor control. Depletion of TRM-like cells abrogated breast tumor protection in parous mice, while enhanced IL-2Rβ signaling-induced TRM-like cells and conferred protection in nulliparous mice. Together, we show a mechanism by which pregnancy can induce anticipatory breast cancer protection in a tissue-specific manner.

As an aggressive subtype of breast cancer, triple-negative breast cancer (TNBC) displays limited therapeutic options and poor clinical prognosis. Thus, there is an urgent need for developing more effective treatment strategies to optimize the clinical management of TNBC. Dihydroartemisinin (DHA), one kind of artemisinin derivatives, exhibits promising antitumor activity mainly through inducing excessive generation of reactive oxygen species (ROS). However, some intrinsic disadvantages, including poor aqueous solubility, short circulation time, and low bioavailability, greatly limit its clinical application. In this research, we constructed an injectable pH/ROS-responsive composite hydrogel (Lip-DHA-CuS@Gel) co-loaded with liposomal DHA (Lip-DHA) and copper sulfide (CuS) nanoparticles to enhance the therapeutic efficacy against TNBC. The hydrogel system could serve as a localized drug depot, enabling sustained drug release and improved tumor retention. In the tumor microenvironment, Lip-DHA-CuS@Gel facilitated efficient intratumoral delivery of DHA and CuS, resulting in mitochondrial copper overload and increased ROS production, thereby synergistically inducing cuproptosis. Copper chelators tetrathiomolybdate (TTM) and D-penicillamine (DPA) could significantly reverse the antitumor effects of Lip-DHA-CuS@Gel in TNBC. In the 4T1 tumor-bearing BALB/c mice, Lip-DHA-CuS@Gel synergistically potentiated the antitumor immunity of PD-1 blockade by inducing the release of damage-associated molecular patterns (DAMPs) and promoting intra-tumoral infiltration of CD8⁺ T cells. Taken together, these findings suggested that Lip-DHA-CuS@Gel highlights significant therapeutic potential to induce cuproptosis and enhance immunotherapy responsiveness in TNBC.

Tumor-infiltrating nerves play critical roles in promoting tumor growth and progression; however, the mechanisms that drive tumor innervation remain unclear. Upon transformation, tumors recruit surrounding peripheral nerves into the tumor microenvironment (TME) to obtain their own innervation, a process called axonogenesis. While in vitro studies suggest tumor cell-derived neurotrophins, such as brain-derived neurotrophic factor (BDNF), drive axonogenesis, this has yet to be demonstrated in vivo. During wound healing, macrophages are the primary source of neurotrophins. Given the critical role of macrophages in breast cancer growth, we investigated whether these immune cells drive tumor axonogenesis in breast tumors in vivo. Syngeneic Py230 mouse triple-negative breast cancer (TNBC) cells were transplanted into intact mice and mice lacking immune-derived BDNF. Bone marrow-derived macrophages from either wild-type or immune-BDNF-deficient mice were transplanted into tumor-bearing recipients to determine if macrophage-derived BDNF was sufficient to restore tumor growth and innervation. We found that transplanted TNBC cannot grow in the absence of immune-derived BDNF, and that depletion of macrophages from the TME compromises tumor innervation. Remarkably, the introduction of wild-type macrophages restores tumor growth and innervation in mice lacking immune-derived BDNF, demonstrating that macrophages are both necessary and sufficient for tumor axonogenesis in vivo. In the absence of sensory tumor innervation, tumor growth was significantly reduced. Moreover, targeting BDNF signaling diminished TNBC growth and innervation. Our findings identify macrophages as the critical source of BDNF driving axonogenesis in breast cancer and suggest that selectively targeting BDNF signaling could provide a novel therapeutic strategy for treating TNBC through compromising tumor innervation.

Breast cancer (BC) metastasis remains a major cause of mortality, yet the molecular mechanisms driving this process are incompletely understood. This study identifies TMEM216, a transmembrane protein implicated in ciliary homeostasis, as a suppressor of lung metastasis in BC. Using mammary-specific Tmem216 knockout mice, we demonstrate that Tmem216 deficiency promotes lung metastasis without affecting primary tumor proliferation. Clinical analyses reveal reduced TMEM216 expression in metastatic lesions and aggressive cell lines, correlating with poor patient distant metastasis-free survival. Mechanistically, TMEM216 interacts with IGF1R and binds to IRS4 via the conserved K79-D1049 interaction, disrupting the IGF1R-IRS4 complex formation and suppressing IGF pathway activation. Rescue experiments in vitro and in vivo confirm that TMEM216-mediated metastasis inhibition depends on IGF signaling modulation. Tissue microarray analyses further establish an inverse correlation between TMEM216 levels and IGF1R phosphorylation in BC patients, with low TMEM216 expression associated with advanced metastasis. These findings delineate TMEM216 as a critical regulator of the IGF1R-IRS4 axis, offering therapeutic opportunities for targeting metastatic BC.

Vasculogenic mimicry (VM) in breast cancer is recognized as an important non-endothelium-dependent vascular mechanism that promotes tumor invasiveness, survival, and metastatic potential; however, effective and specific intervention strategies remain limited. In this study, we innovatively constructed a nanodelivery system based on hybrid membranes derived from bacterial outer membrane vesicles and breast cancer cell membranes, termed mTOMV, and loaded it with the natural flavonoid isoliquiritigenin (ISL) to enhance targeted antitumor activity. ISL@mTOMV nanoparticles exhibited a uniform spherical morphology, favorable stability, and good biocompatibility. In vitro experiments demonstrated that ISL@mTOMV significantly inhibited the proliferation, migration, and invasion of MDA-MB-231 and MCF-7 breast cancer cells, while promoting tumor cell apoptosis. Mechanistically, ISL@mTOMV markedly reduced the expression of ACLY, H3K27ac, and SMAD5 and suppressed the activity of the TGF-β/SMAD signaling pathway. In vivo experiments further confirmed that ISL@mTOMV inhibited ACLY-mediated H3K27 acetylation, attenuated SMAD5 transcriptional activation, and blocked TGF-β/SMAD signaling, thereby significantly suppressing VM formation in breast cancer. Collectively, this study proposes a highly biocompatible and tumor-targeted nanotherapeutic strategy, providing a new technical approach and potential translational value for VM intervention in breast cancer.

Breast cancer screening and longitudinal monitoring require imaging technologies that are portable, operator-independent, and suitable for frequent use, a capability not fully met by mammography or conventional ultrasound. We present a three-dimensional (3D) portable ultrasound system for real-time examination (3D PURE) that overcomes key limitations in volumetric breast imaging through advances in transducer design, acoustic materials, and adaptive beamforming. A box-array design incorporating a corner-gap offset geometry suppresses peak crosstalk (by 3.73 dB at the corner-most element), prevents preamplifier saturation, and supports higher transmit voltages (up to 24 V). A custom flowable backing layer (impedance 6.12 MRayl; attenuation 7.56 dB mm-1 MHz-1) integrates around fragile wirebonds, reduces inter-element crosstalk by ~4.5 dB throughout the array, and improves axial and lateral/elevational resolutions by ~200 µm and ~70 µm, respectively. Layered Aberration-Correction Reconstruction (LACR), an adaptive 3D beamformer, compensates for heterogeneous speed-of-sound (SoS) in the breast, reducing depth localization error by 2 mm and aberration defocusing by 70 µm on average at a 5 cm depth. Nine of the ten participants in an in vitro study showed improved microtarget detection efficiency with 3D PURE relative to a conventional 2D system (p = 0.0215), analogous to detection of microcalcifications. These results, combined with in vivo imaging validation of various phenotypes, highlight the potential of 3D PURE for reliable breast imaging. Furthermore, a vision-guided computer interface, MyFUS, ensures self-guided, user-friendly, and operator-independent probe positioning for longitudinal monitoring.

Nitric oxide synthase 2 (NOS2) and cyclooxygenase 2 (COX2) tumor expression present significant obstacles for effective treatment of aggressive tumors including ER-negative breast cancer. Spatial analysis of NOS2, COX2 and CD8 expression in patient tumors has identified mechanisms of treatment inhibition that were further elucidated using the 4T1 mouse model of triple negative breast cancer and live cell culture studies. NOS2 and COX2 activate each other via a feed-forward paracrine mechanism. While NOS2 promotes cancer stemness and the formation of metastatic niches, COX2 mediates CD8+ T cell suppression. Quantitative spatial analysis has revealed NOS2/COX2 roles during the temporal progression from immune hot in low COX2 expressing tumors to three immune cold stages in the tumor microenvironment of high COX2 expressing tumors. Type 1: immune cold with stroma-restricted CD8+ T cell secretion of interferon gamma, which activates COX2 expression at the tumor margin as well as NOS2 expression at the tumor periphery. Type 2: developing immune desserts lack stroma-restricted CD8+ T cells with tumor NOS2 and COX2 restricted to the tumor periphery. Type 3: mature immune desserts exhibit abated NOS2, COX2 and CD8+ T cells, and induction of B7H4 and cancer-associated fibroblasts driven by significant tumor hypoxia and necrosis. These three types of immune desserts can coexist with each other and with immune hot regions in the same tumor. The coordinated interplay of NOS2 and COX2 indicates that targeting both these enzymes provides an effective treatment strategy that is supported by ongoing clinical trials demonstrating improved clinical outcomes in patients who have otherwise exhausted treatment options.

Triple-negative breast cancer (TNBC) exhibits hyperactive EGF (epidermal growth factor) signaling that drives metabolic plasticity and metastasis. Here, we identify secretory macroautophagy/autophagy as a key downstream effector linking EGF signaling to metabolic reprogramming that fuels TNBC metastatic progression. In TNBC cells, EGF stimulation redirected autophagosomes toward the plasma membrane through a SEC22B-dependent route, signifying activation of secretory autophagy. Proteomic profiling of purified autophagosomes revealed enrichment of the lactate transporter SLC16A3/MCT4 and its chaperone BSG/CD147 on autophagosomal membranes. Mechanistically, EGF promoted MAP1LC3/LC3-SLC16A3 interaction, facilitating SLC16A3 trafficking to the plasma membrane and enhancing lactate efflux. Genetic or pharmacological blockade of autophagy abrogated SLC16A3 surface localization, reduced extracellular lactate accumulation, and markedly suppressed lung metastasis originating from orthotopic TNBC tumors in mice. Although pharmacological inhibition of SLC16A3 effectively blocks its transporter activity and reduces lactate secretion, targeting autophagy provides a more precise approach to suppress EGF-driven SLC16A3 expression and the consequent rise in lactate secretion. Clinically, multiplex immunofluorescence of patient tumors demonstrated strong co-expression of EGFR, LC3, and SLC16A3, which correlated with poor disease-free survival. Our study reveals a previously unrecognized EGF-secretory autophagy axis that orchestrates metabolic remodeling in TNBC and highlights the therapeutic potential of targeting the secretory autophagy- SLC16A3-lactate pathway to restrain metastasis.Abbreviations: 3-MA: 3-methyladenine; ATG5: autophagy related 5; ATG7: autophagy related 7; APf: autophagosome fraction; CQ: chloroquine; CRISPR-Cas9: clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9; EGF: epidermal growth factor; EGFR: epidermal growth factor receptor; ER: endoplasmic reticulum; ERBB2/HER2: erb-b2 receptor tyrosine kinase 2; GOBP: gene ontology biological process; imBI: induced metabolic bioluminescence imaging; i.p.: intraperitoneal injection; IVIS: in vivo imaging system; LAMP2: lysosomal associated membrane protein 2; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MAPK/ERK: mitogen-activated protein kinase; PLA: proximity ligation assay; PNS: postnuclear supernatant; SEC22B: SEC22 homolog B, vesicle trafficking protein; shRNA: short hairpin RNA; SLC16A3/MCT4: solute carrier family 16 member 3; SNARE: soluble N-ethylmaleimide-sensitive-factor attachment protein receptor; TIRF: total internal reflection fluorescence; TME: tumor microenvironment; TNBC: triple-negative breast cancer; ULK1/Atg1: unc-51 like autophagy activating kinase 1.

Tissue-level phase transitions are emerging as a crucial mechanism in tumour development and metastasis. When becoming invasive, epithelial tumours undergo a transition from a solid-like state to a more fluid-like one. Although the contributions of cell adhesions, traction forces and cell migration for such behaviour are known, the exact biophysical and molecular mechanisms controlling these transitions are not fully understood. Here we show that breast cancer cell fluidity is regulated by IRSp53, a protein linking plasma membranes to the cytoskeleton. In both two-dimensional monolayers and three-dimensional spheroids, the depletion of IRSp53 increases fluidity and active wetting of the substrate due to a decrease in intercellular friction and enhanced local cell rearrangements. Molecularly, IRSp53 interacts with the junctional protein Afadin to control global tensile state and active wetting, establishing these proteins as key regulators of epithelial collectives' viscosity in breast cancer tumouroids. In breast cancer patient samples, low IRSp53 expression levels and aberrant localization correlate with worse clinical outcomes. These findings support the broader relevance of IRSp53-regulated mechanics in epithelia and their potential prognostic value in cancer.

Cancer cells that seed in the lung require lipids often produced by alveolar type II (AT2) cells. However, whether overt metastases depend on AT2 cell-derived lipids and whether AT2 cells can be targeted to reduce metastasis growth remains unknown. We discovered that breast cancer-derived lung metastases stimulate the proliferation of AT2 cells in their vicinity and reprogram them into lipid feeder cells in mice and patients using spatial analysis. Mechanistically, the metastasis secretome activates the transcription factor sterol regulatory element-binding transcription factor 1 (SREBP-1) in AT2 cells, enhancing the expression of key de novo lipid synthesis genes, including fatty acid synthase (FASN) and glycerol-3-phosphate acyltransferase 1 (GPAM). Deleting Fasn selectively in AT2 cells or targeting FASN and GPAM systemically significantly impairs lung metastasis growth in mice. In summary, we discovered that overt metastases reprogram AT2 cells and that targeting the lipid metabolism of AT2 cells impairs metastasis growth. Current therapies in oncology targeting the cancer or immune cell compartment of tumors show limited efficacy against breast cancer-derived metastases. We discovered that decreasing the lipid metabolism of lung resident AT2 cells is sufficient to impair lung metastasis growth in mice without apparent adverse effects.

Cancer progression is driven by the dynamic interplay between metabolic reprogramming and immune evasion. A central mechanism is aerobic glycolysis, which fuels tumor growth while simultaneously impairing antitumor immunity. To address recurrent metastatic triple-negative breast cancer (TNBC), we developed a biomimetic nanoplatform (3BP@CP NPs) composed of high-affinity programmed death-1 (PD-1)-modified cell-membrane nanovesicles encapsulating 3-bromopyruvate (3BP)-loaded nanoparticles. The optimized nanoparticles exhibit enhanced pharmacokinetics with prolonged circulation, enabling dual programmed death-ligand 1 (PD-L1)-targeted tumor homing and checkpoint inhibition. The glycolytic inhibitor 3BP specifically inhibits hexokinase II (HK2) activity, triggering metabolic collapse and immunogenic cell death while reversing immunosuppression in the tumor microenvironment (TME). This synergistic metabolic-immunological intervention elicits robust systemic antitumor responses, curtailing tumor recurrence and metastasis while extending survival in aggressive TNBC models. Collectively, this study establishes a therapeutic paradigm combining immune checkpoint receptor-modified cell-membrane nanovesicles (ICB CVs) with metabolic modulators to enhance immunotherapy efficacy in recurrent metastatic TNBC, providing a clinically translatable approach for PD-L1-expressing malignancies.

Radiotherapy (RT) is a cornerstone of cancer treatment, but its efficacy is often compromised by robust antioxidant defense mechanisms that counteract radiation-induced oxidative stress. In this study, we developed a novel dual-action nanoplatform, termed radio-activatable lipid nanoparticles (RaLNPs), designed to enhance radiosensitivity by amplifying radiation-induced ferroptosis. RaLNPs incorporate both siRNA targeting glutathione peroxidase 4 (siGPX4), a key ferroptosis defense antioxidant enzyme, and 7-dehydrocholesterol (7-DHC), a radiation-reactive lipid. Notably, the structural lipid cholesterol was completely replaced with 7-DHC, thereby designing the carrier itself to possess a therapeutic function activated by irradiation. The engineered RaLNPs exerted a dual-action mechanism by suppressing GPX4 expression to disable the ferroptosis defense system and, upon irradiation, amplifying 7-DHC-mediated radical chain reactions. Importantly, RaLNPs did not induce oxidative stress or ferroptosis in the absence of radiation, whereas therapeutic irradiation selectively triggered potent and iron-dependent ferroptosis. Beyond direct tumor cell killing, this ferroptotic process also elicited the key hallmarks of immunogenic cell death (ICD), thereby promoting dendritic cell maturation. In a syngeneic 4T1 breast cancer mouse model, the combination of RaLNPs and a single dose of radiation exhibited superior suppression of primary tumor growth and was accompanied by a reduction in metastatic lesions, without systemic toxicity. Analysis of tumor tissues revealed that this therapeutic efficacy was driven by a coordinated immune response, linking T-cell priming in tumor-draining lymph nodes to the sustained intratumoral infiltration of functional cytotoxic T lymphocytes. In conclusion, the RaLNPs developed in this study act as innovative radio-activatable radiosensitizers that simultaneously induce tumor cell death and antitumor immunity specifically in response to irradiation. This work highlights a transformative strategy in which a conventional lipid nanoparticle carrier is evolved into an active therapeutic to overcome the limitations of radiotherapy.

Ferroptosis, a promising therapeutic strategy for triple-negative breast cancer (TNBC), faces significant challenges due to intrinsic tumor defense mechanisms. To enhance ferroptosis against TNBC, a biomimetic "four-in-one' cascade nanozyme AuPd/Cu2O@Cancer cell membrane (APCM) was engineered to remodel tumoral redox microenvironment and disrupt cancer cell energy metabolism. APCM nanozyme integrates four enzyme-mimicking activities into a single nanoplatform, including peroxidase-like, glucose oxidase-like, catalase-like, and glutathione peroxidase-like. This synergistic cascade converts endogenous H2O2 to cytotoxic ·OH, depletes glucose to block energy supply while self-supplying H2O2, alleviates hypoxia, and depletes glutathione to suppress antioxidant defense, collectively triggering lethal reactive oxygen species (ROS) accumulation for ferroptosis. APCM further enables photothermal therapy (PTT), inducing direct thermal ablation and providing localized heat to augment nanocatalytic efficacy. Coating with tumor-derived membrane facilitates homologous targeting and immune evasion. Transcriptomic analysis confirmed profound APCM-mediated modulation of ferroptosis, metabolic, and redox-associated gene signatures. Notably, the APCM nanozyme enables dual-mode imaging, offering visualization of the location of TNBC and precise guidance for treatment. Collectively, this "four-in-one' biomimetic nanozyme, which integrates multiple enzyme-mimicking activities and tumor-cell-membrane camouflage, effectively disrupts redox and metabolic homeostasis to potentiate ferroptosis, establishing a promising therapeutic paradigm for TNBC.

Breast cancer (BC) treatment efficacy is often compromised by tumor cell plasticity and multidrug resistance of multi-factorial origin. Among emerging therapeutic agents, flavonoids - a structurally diverse group of naturally occurring polyphenols - have demonstrated a significant potential to modulate the Janus kinase/signal transducer and activator of transcription (JAK-STAT) signaling pathway, a key driver of oncogenic transformation, immune evasion, and therapeutic failure in BC. Notably, natural compounds, including but not restricted to apigenin, quercetin, naringenin, morin, luteolin, butein, xanthohumol, silibinin, baicalein, nobiletin, rosmanol, orientin, eriocitrin, breviscapine, 8-hydroxydaidzein, icariside I, cardamonin, epigallocatechin-3-gallate, fisetin, brutieridin, melitidin, isoliquiritigenin, and sophoraflavanone G, have been recognized as potent JAK-STAT modulators in BC models. Through targeted interference with this pathway, flavonoids exert pleiotropic antitumor effects - enhancing the efficacy of chemotherapeutic agents, suppressing cellular proliferation and invasion, and reducing tumor aggressiveness. Beyond JAK-STAT signaling, flavonoids also influence several additional therapy resistance-related mechanisms, including regulation of ATP-binding cassette (ABC) transporters, promotion of apoptosis, suppression of epithelial-mesenchymal transition (EMT) and cancer stem cells (CSCs), cell cycle arrest, mitochondrial autophagy and mitophagy modulation, as well as remodeling of the tumor microenvironment. This article provides a comprehensive overview of the molecular mechanisms by which flavonoids overcome therapeutic resistance in BC, focusing on JAK-STAT signaling modulation. The paper follows principles of Predictive, Preventive, and Personalized Medicine (3PM), promoting the paradigm shift from reactive to proactive medicine. Contextually, it underscores the importance of multidisciplinary research to elucidate flavonoid-specific mechanisms, identify predictive biomarker panels, and develop advanced delivery systems, integrates patient phenotyping, multi-level diagnostics, and AI-driven data interpretation to enable cost-effective primary and secondary prevention, and therapeutic algorithms tailored to personalised patient profiles improving therefore individual outcomes.

Ferroptosis is a prospective approach for cancer treatment. However, the efficacy of ferroptosis therapy is limited by three parallel ferroptosis defense pathways: the glutathione (GSH)-glutathione peroxidase 4 (GPX4) pathway, the ferroptosis suppressor protein 1 (FSP1)-ubiquinol (CoQH2) pathway, and the dihydroorotate dehydrogenase (DHODH)-CoQH2 pathway. Inspired by the principles of preprogrammed chemical reaction networks (CRNs), herein, a novel drug delivery system (SRF@Au@M NPs) was designed based on MIL-100(Fe) for predictable behaviors in tumor cells to break the three ferroptosis defense systems. SRF@Au@M NPs were fabricated through the size optimization of MIL-100(Fe), sorafenib (SRF) loading, in-situ growth of Au nanoparticles (Au NPs) and surface modification with dihydrolipoic acid derivatives. SRF@Au@M NPs disintegrate in the presence of high concentrations of GSH, releasing sorafenib (SRF) into tumor cells, which reduces GSH synthesis and inhibits GPX4 activity. The Au nanoparticles decompose glucose to produce H2O2, providing fuel for the Fenton reaction and disrupting carbohydrate metabolism to inhibit NAD(P)H generation. Particularly, a novel redox-CRN was formed between dihydrolipoic acid derivatives and iron ions, continuously promoting reactive oxygen species generation while concurrently consume NADH. The imbalance of NAD(P)H metabolic homeostasis impedes the recycling of CoQ to CoQH2, resulting in the simultaneous inhibition of the FSP1-CoQH2 and DHODH-CoQH2 pathways. Consequently, the SRF@Au@M NPs triggered a potent ferroptosis storm in 4T1 tumor cells and achieved an 92.5% tumor growth inhibition in tumor-bearing mice, significantly higher than that of other treatment groups. Our sophisticated strategy based on CRNs provides a new promising paradigm for ferroptosis activation and cancer treatment.

Breast cancer is a biologically heterogeneous disease in which tumor-intrinsic diversity and the tumor immune microenvironment jointly shape therapeutic resistance and variable clinical outcomes. Although nanomedicine has improved the safety and pharmacokinetic profiles of several anticancer agents, clinically approved nanocarriers have produced limited efficacy gains, partly because of heterogeneous tumor accumulation, restricted penetration, and empirical formulation design. Polymeric lipid nanoparticles (PLNs), also known as lipid-polymer hybrid nanoparticles, provide a tunable core-shell platform that combines the structural stability of polymeric systems with the biomimetic and functional versatility of lipid-based carriers. These properties enable controlled drug loading, adjustable release kinetics, and surface engineering for targeting or immune modulation. Artificial intelligence (AI) may support PLN development by organizing complex formulation variables and prioritizing experimentally testable designs rather than replacing mechanistic nanobiology. Machine learning, graph-based models, generative approaches, and predictive pharmacokinetic frameworks can help connect biological barriers, including receptor heterogeneity, stromal restriction, immune contexture, and delivery variability, with modifiable formulation parameters such as particle size, lipid-polymer composition, ligand density, and release behavior. Microfluidic manufacturing may further improve reproducibility by translating computationally prioritized formulations into controlled physical nanoparticles. This review summarizes the structural rationale and functional advantages of PLNs in breast cancer, evaluates barrier-oriented PLN design strategies, and examines the role of AI in formulation optimization, biological fate prediction, drug-release modeling, and translational workflow design. We also discuss current limitations, including data scarcity, limited PLN-specific validation, clinical delivery heterogeneity, and regulatory challenges. Overall, AI-guided PLN development should be viewed as a biology-informed and manufacturing-aware framework for improving formulation prioritization and reproducibility, rather than as an immediate clinical solution.

In estrogen receptor-positive (ER +) breast cancer, CDK4/6 inhibitors (CDK4/6is) combined with endocrine therapy (ET) are standard first-line treatment for metastatic disease. However, most patients eventually develop resistance. Activating ESR1 mutations are a prevalent mechanism of acquired resistance to ET and are enriched after ET plus a CDK4/6 inhibitor (CDK4/6i), but their role in the clonal evolution and adaptive mechanisms of acquired resistance to CDK4/6 inhibition, independent of ET, is unknown. In addition, whether different CDK4/6is impose distinct selective pressures and divergent resistance states remains elusive. To investigate the clonal dynamics, cell states and cellular plasticity during acquired CDK4/6i resistance in mutant versus wild-type (WT) ESR1, we performed high-complexity DNA barcoding (ClonTracer library) with longitudinal sampling and multi-omic profiling in an isogeneic MCF7 model expressing WT ER or Y537S mutant ER. We also evaluated the clonality of the ESR1 mutations in clinical samples with CDK4/6i resistance. We showed that ESR1 mutations are enriched in clinical tumors with acquired resistance to CDK4/6is, and in paired biopsies expanded to near clonality after treatment. We demonstrated progressive clonal selection with both divergent and partially convergent evolutionary trajectories. The ESR1 mutation substantially reshapes clonal and epigenetic evolution during palbociclib resistance but had a weaker impact under abemaciclib selection. Overall, clonal evolution and cell states in palbociclib and abemaciclib resistance were distinct. Single-cell RNA-seq revealed transcriptional heterogeneity highlighting cellular plasticity during passaging of cells and selection. Finally, in vivo barcoding of mammary xenograft, local recurrences, and distant metastases demonstrated site-specific clonal outgrowth in mutant ER metastases, and partial overlap between metastatic and CDK4/6i-resistant subclones, supporting the dual role of specific populations in therapeutic resistance and metastatic colonization. High-resolution lineage tracing and multi-omic studies demonstrate that CDK4/6i resistance is shaped by clonal selection and adaptive remodeling of cell states, with the ESR1 mutation status and the specific inhibitor acting as key determinants of evolutionary trajectories. These findings suggest that both variables should be considered when designing sequential and combination treatment strategies to overcome CDK4/6i resistance.

Pancreatic ductal adenocarcinoma (PDAC) accounts for 90% of pancreatic cancers and has a very poor prognosis. Ten to 15% are staged as resectable at diagnosis, and 5% to 15% downstaged with therapy to where surgery is feasible. Chemotherapy is a mainstay for all stages of PDAC. Targeted therapies are available for patients with select but expanding actionable genomic alterations. The tumor microenvironment provides a dense stroma with an immunosuppressive milieu that contributes to inherent treatment resistance of PDAC. Herein, we review current management of PDAC with a focus on emerging treatment paradigms, including targeted and immunomodulatory agents. PDAC is a complex disease with unique genomic, immunologic, and clinical features. Recent developments in understanding of the pathobiology of this disease are translating into targeted and immunomodulatory therapies that will alter treatment paradigms and improve outcomes for this recalcitrant malignancy.

We present 18 F-FAPI-74 PET/CT findings before and after proton beam therapy (PBT) in a 71-year-old woman with liver metastasis following pancreatic cancer resection. Before PBT, no uptake was observed on FDG-PET, whereas the lesion was visualized on FAPI-PET, consistent with contrast-enhanced MRI. Follow-up FAPI-PET at 3 and 6 months post-PBT revealed mild, newly developed uptake confined to the high-dose irradiated area. However, contrast-enhanced MRI demonstrated complete tumor disappearance and normalization of serum tumor marker levels. These findings suggest that post-irradiation 18 F-FAPI-74 uptake is more likely attributable to reactive changes, such as radiation-induced hepatic fibrosis, rather than residual or recurrent tumor.

Pancreatic ductal adenocarcinoma (PDAC) growth and metastasis are influenced by the tumor microenvironment (TME), which includes immune cells, endothelial cells, macrophages, and cancer-associated fibroblasts (CAFs). Since the novel integrin-targeted cytotoxin ProAgio can inhibit activated CAFs and endothelial cells, we investigated its combination with standard of care chemotherapies in genetically engineered mouse (GEM) and orthotopic murine models of PDAC. We established metastatic murine KPC-Ganji & Bassel-Luc (mKPC-GB-Luc) cell lines and validated them using bulk RNA sequencing. Two in vivo orthotopic mouse model were used to evaluate ProAgio with chemotherapy. mKPC-GB-Luc was used to evaluate the 5FU, oxaliplatin, and irinotecan (FOI combination), and KPC-ML1-Luc was used to evaluate the gemcitabine, nab-paclitaxel (GPTx combination). Immunohistochemistry was used to measure integrin β3, E-cadherin, and HIF-1α. Hypoxia was evaluated using pimonidazole. Stem cells, CAFs, and immune cell subtypes were quantified by flow cytometry. We evaluated the combination of ProAgio plus gemcitabine in a KPC genetically engineered mouse model (KPC GEM). Two sets of KPC GEM were developed. The first set was used for a survival study. The second set was terminated early and tumors were used for single-cell RNA seq and sequential multiplex immunofluorescence (COMET). Compared with the chemo regimen (GPTx or FOI), ProAgio, or sham, the combination of ProAgio plus chemo significantly modulated the TME, reduced tumor weight, reduced hypoxia, and eliminated metastasis to the lungs and liver. The combination of ProAgio and gemcitabine increased overall survival in KPC GEM mice compared with either treatment alone. Single-cell RNA sequencing, flow cytometry, immunofluorescence, immunohistochemistry, and COMET analyses demonstrated that ProAgio plus chemotherapy reprogrammed the PDAC-TME by changing activated CAFs toward a qCAFs (quiescent CAFs), macrophages from protumor to proinflammatory polarization, and activating natural killer (NK) cells, CD4⁺, and CD8⁺ T cells. Combination therapy inhibited PDAC stemness. ProAgio-treated CAFs in vitro exhibited reduced secretion of glycine and cysteine, vital metabolites that support stemness and tumor progression. These results confirm the novel mechanism of action of ProAgio, which includes reducing hypoxia and modulating TME. The current data provide evidence for potentiation of chemotherapy efficacy by ProAgio.

As key constituents of cellular sphingolipid pools, sphingomyelin (SM) and ceramide (CER) are central to the regulation of cancer cell death and survival. The metabolic flux between these two lipids is a vital component of the cellular stress response, yet the underlying regulatory mechanisms in cancer remain elusive. Acid sphingomyelinase (SMPD1) facilitates the conversion of SM to CER, functioning as a key enzymatic driver of CER-mediated signalling. Herein, we aim to evaluate the role of SMPD1-driven sphingolipid metabolism in pancreatic carcinogenesis. A targeted quantitative analysis of the plasma metabolome was conducted involving patients with pancreatic ductal adenocarcinoma (PDAC, n=202) and matched control subjects (n=204). Multiplex immunohistochemistry was performed on resected PDAC (n=122) to identify expression of SMPD1 with tumour and immune cell markers. CRISPR/Cas9 driven Smpd1-deleted murine cell lines were generated and subsequently assessed for their carcinogenic potential in vitro. The effects of Smpd1 deletion on tumour formation were evaluated using both syngeneic orthotopic and metastatic murine models. Here, we demonstrate that tumour cell-autonomous expression of SMPD1, in pancreatic ductal adenocarcinoma (PDAC), is associated with poorer patients' outcomes. Smpd1 ablation in murine PDAC cells resulted in reduced proliferation and migration in vitro and decreased metastases and tumour burden in vivo. Integrated transcriptomic, metabolomic and proteomic studies revealed that SMPD1 abrogation impairs KrasG12D oncogenic signalling and, thus, reduces tumour burden. Reduced plasma membrane interaction of KrasG12D was associated with SMPD1-dependent sphingolipid metabolism. Notably, the SMPD1 inhibitor (ARC39) potently synergised with the KrasG12D inhibitor (MRTX1133). In summary, SMPD1 regulated plasma membrane sequestration of KrasG12D represents a potential therapeutic target within the Kras signalling pathway for intractable PDAC.

Fibroblast activating protein (FAP) expressing fibroblasts are an attractive target for cancer therapeutic depletion and while preclinical depletion shows success, previous modalities have had unsuccessful clinical impact. Here, we wanted to comprehensively understand the tumor microenvironmental changes after FAP+ fibroblast depletion and unravel potential reasons for resistance and vulnerabilities that appear upon treatment with a FAP-targeted T-cell engager. To unveil the complex changes that occur in the tumor microenvironment (TME) after FAP+ fibroblast depletion, we generated comprehensive single-cell RNA-sequencing analysis of FAP+ cancer-associated fibroblasts (CAFs) depletion within the TME to understand the key populations and genetic modulations in all cell subtypes. A CD3 T-cell engager directed against FAP (FAP TcE) was used to deplete FAP+ fibroblasts in a preclinical murine model of pancreatic cancer. To understand complex population dynamics on FAP TcE, we performed single-cell RNA-sequencing of treated versus untreated tumors to unveil population and genetic changes. Administration of FAP TcE resulted in tumor growth control in vivo that was not dependent on T-cell priming and egress via draining lymph nodes. After FAP TcE, T-cell exhaustion was prevalent with an increased T-cell exhaustive state and the emergence of a T-cell progenitor exhausted state, yet the addition of anti-programmed cell death protein 1 (PD-1) failed to enhance tumor efficacy. Within fibroblast populations, FAP TcE depleted FAP+ CAFs; however, depletion is compensated by an emergence of a "mixed CAF" population, potentially limiting the efficacy of FAP TcE. This study highlights the complex and plastic fibroblast changes occurring with stroma-targeted therapies that may limit therapeutic efficacy.

The application of the novel KRASG12D inhibitor in pancreatic ductal adenocarcinoma (PDAC) is currently hindered by adaptive resistance. Metabolic reprogramming is a hallmark of KRASG12D signalling, yet the mechanisms linking these alterations to immunosuppression and low therapeutic response are poorly defined. To identify the key regulatory nodes connecting KRASG12D-driven metabolic adaptations to tumour microenvironment and develop a mechanistic-based combinatorial strategy. We integrated whole-exome sequencing, untargeted metabolomics and single-cell RNA sequencing of human PDAC specimens to analyse the metabolic-immune landscape. We evaluated therapeutic efficacy using the autochthonous mouse and patient-derived xenograft models. We found that KRASG12D enhanced cholesterol metabolism and promoted CD8+ T cell exhaustion, whereas KRASG12D inhibition or cholesterol synthesis blockade induced compensatory ULK1-associated autophagy. Cotargeting cholesterol metabolism and autophagy potentiated the antitumour efficacy of the KRASG12D inhibitor MRTX1133 and alleviated CD8+ T cell exhaustion. Mechanistically, KRASG12D transcriptionally upregulated USP20 via EGR1, which simultaneously deubiquitinated and stabilised 3-hydroxy-3-methylglutaryl-CoA reductase and ULK1, thereby orchestrating cholesterol metabolism and autophagy-associated survival. Genetic depletion or pharmacological inhibition of USP20 with GSK2643943A suppressed these pathways and restored CD8+ T cell function, improving responses to MRTX1133 and anti-programmed cell death protein-1 (anti-PD-1). In preclinical PDAC models, triple therapy with GSK2643943A, MRTX1133 and anti-PD-1 elicited a robust therapeutic response and induced significant tumour regression. USP20 acts as a critical metabolic checkpoint that orchestrates CD8+ T cell exhaustion and therapeutic response. Targeting the USP20-cholesterol-autophagy axis represents a promising strategy to reverse immune suppression and unlock the full potential of KRASG12D inhibitors in PDAC.

Min, Schweizer, and colleagues use artificial intelligence-powered deep visual proteomics to generate a spatial proteomic atlas of pancreatic cancer precursor evolution, revealing that major metabolic and inflammatory reprogramming occurs long before overt histologic transformation. More broadly, the study highlights the emerging potential of spatial proteomics and multiomics to bridge histopathology with molecular pathology and precision oncology. See related article by Min et al., p. 1323.

Pancreatic cancer (PaC) remains one of the deadliest cancers, with 5-year survival rates of 13%. A major driver of its aggressiveness is the tumour microenvironment (TME), which fuels tumour growth, metastasis, and therapeutic resistance through dynamic, bi-directional communication between cancer cells, fibroblasts, and immune cells. Emerging evidence highlights extracellular vesicles (EVs) as key mediators of oncogenic cross-talk within the PaC TME. This study demonstrates for the first time that the overexpression of metastasis suppressor N-myc downstream regulated gene 1 (NDRG1) significantly influences the biogenesis, cargo packaging and release of EVs by cancer cells. This was mediated by a direct interaction between NDRG1 and ALIX, a key protein involved in EV biogenesis and packaging, with NDRG1 facilitating ALIX proteasomal degradation. Further, EVs released from NDRG1-overexpressing cells had significantly fewer CAF-activation proteins (i.e. TGF-β), leading to attenuated ERK1/2 and p38 activation in pancreatic stellate cells (PSCs), and reduced expression of key fibrotic markers (α-SMA, FAP, and collagen 1A). NDRG1 overexpression also reduced sEVs uptake by PaC cells and diverted these to the lysosome for degradation. These findings uncover a previously unrecognized mechanism by which NDRG1 overexpression disrupts the oncogenic two-way communication between PaC cells and the TME, positioning NDRG1 overexpression as a compelling therapeutic approach against this formidable malignancy.

Fibrosis and tumour innervation are two features of the tumour microenvironment (TME) that contribute directly to the lethality of pancreatic ductal adenocarcinoma (PDAC), but their potential interactions have not been explored. Moreover, although it is known that activated Schwann cells (SCs) stimulate cancer cell invasion, it remains unclear how SCs are activated. We determined how SCs are activated in the pancreatic fibrotic microenvironment. The correlation between physical features of the microenvironment and SC activation was assessed in human patient samples and in mice by SC c-Jun phosphorylation monitoring, atomic force microscopy and multiphoton live imaging. Several in vitro models in which forces were applied to SCs expressing a reporter for c-Jun phosphorylation and RNA-Seq analysis were used to decipher the cellular and molecular mechanisms of SC activation. Nerves surrounded by stiff stroma present higher SC activation. Intravital imaging shows a matrix-dependent SC activation. Mechanical forces on SCs induce c-Jun phosphorylation in SCs in a non-canonical manner that involves a nuclear sensing machinery with the pro-inflammatory enzyme phospholipase A2. Fibrosis enhances the protumorigenic impact of innervation by activating SCs via a mechanism in which nuclear compression triggers non-canonical activation of the AP-1 transcription factor complex. Pancreatic fibrosis alone, without cancer cells, is sufficient to activate SCs, suggesting this mechanism may be common across non-malignant pancreatic diseases. Notably, SCs are more sensitive to mechanical activation than PDAC cells. These findings reveal TME interactions that may guide future microenvironment-targeted PDAC therapies.

Pancreatic ductal adenocarcinoma (PDAC) evolves through precursors, yet the protein programs governing early progression remain poorly defined. We applied Deep Visual Proteomics (DVP)-integrating computational pathology, laser microdissection, and mass spectrometry (MS)-to profile normal ducts, acinar-to-ductal metaplasia (ADM), low-grade (LG) and high-grade (HG) pancreatic intraepithelial neoplasia (PanIN), and invasive carcinoma from organ donors and patients with PDAC. Quantifying 9,181 proteins from ∼100 cells per region, we uncovered a molecular field effect in histologically normal ducts and proteomic divergence of LG-PanINs by cancer context. We identified four stage-associated molecular programs. Stress adaptation and immune engagement emerged early in cancer-associated normal ducts. Metabolic reprogramming initiated in normal ducts and intensified across PanIN progression. Mitochondrial remodeling became prominent in HG-PanINs before invasion. MS detected KRAS hotspot mutant peptides within incidental precursor lesions from cancer-free individuals. These findings demonstrate that molecular reprogramming precedes histologic transformation, creating opportunities for earlier detection of lethal cancer. Artificial intelligence (AI)-guided DVP represents the first in-depth assessment of the proteomic landscapes observed during the multistep progression of pancreatic adenocarcinoma, including histologically normal ducts, ADM, and LG- and HG-PanIN lesions. These data represent a unique resource of candidate biomarkers and interception targets against this lethal disease. See related commentary by Yang and Fan, p. 1255.

Chimeric antigen receptor (CAR) T cells have shown great potential in hematological cancers, but lack efficacy in solid tumors, highlighting the need for novel strategies. Stimulator of interferon genes (STING) activation was shown to inflame the tumor microenvironment, but combination of STING agonists and CAR-T cells might be limited by detrimental outcomes of T cell-intrinsic STING activation. In this study, we evaluated the potential of combining STING agonists and CAR-T cells in the context of pancreatic cancer. We assessed the synergy of CRISPR-Cas9-edited CAR-T cells and the STING agonist diABZI within a T cell exhaustion model in vitro and both xenograft and syngeneic mouse models in vivo. Combination of STING-ablated CAR-T cells and diABZI resulted in enhanced cancer cell killing, increased CAR-T cell proliferation, reduced exhaustion, and expansion of an effector-memory phenotype in vitro. Mechanistically, superior CAR-T cell functionality required genetic ablation of STING in CAR-T cells and was dependent on cancer cell-intrinsic STING signaling on STING-agonistic treatment. Moreover, we identified a synergistic feedback loop comprising the T cell-secreted cytokines interferon-γ and tumor necrosis factor, which prime STING signaling within cancer cells, thereby potentiating the outcomes of cancer cell-intrinsic STING activation in inducing ameliorated CAR-T cell states. Ultimately, we could demonstrate that combination of STING deficient CAR-T cells and diABZI was able to provide enhanced tumor control in both xenograft and syngeneic mouse models. This was accompanied by increased intratumoral CAR-T cell numbers and reprogramming of the tumor microenvironment in vivo. Our findings suggest that STING deficient CAR-T cells stand to benefit from STING agonists to improve CAR-T cell therapy for immune-deprived cancers such as pancreatic cancer.

Cancer-associated fibroblasts (CAFs) are vital in advancing pancreatic ductal adenocarcinoma (PDAC) cancer growth and fibrosis. Although their roles are recognized, the precise molecular mechanisms remain incompletely understood. The study focuses on PINCH-1, a molecule elevated in various tumors, and its lesser-known effects on tumor stromal cells. This work reveals that PINCH-1 ablation in pancreatic CAFs (P-CAFs) significantly reduces extracellular matrix (ECM) expression, particularly collagen I and fibronectin. For collagen I, this effect is due to PINCH-1's ability to stabilize Notch1 protein, preventing Notch1 degradation, which is crucial for collagen I production. Reintroducing PINCH-1 can reverse collagen I reduction, highlighting a novel mechanism for matrix remodeling through the PINCH-1-Notch1 axis. Moreover, PINCH-1 also boosts cancer cell proliferation via the AKT signaling pathway, as demonstrated in both in vitro and in vivo experiments. We identify PINCH-1 as a master regulator within P-CAFs that promotes PDAC progression through dual mechanisms: (1) sustaining desmoplasia via Notch1-dependent collagen I production and (2) fueling cancer cell proliferation via AKT activation. Targeting the PINCH-1/Notch1/AKT axis presents a novel theragnostic strategy to simultaneously disrupt the fibrotic tumor microenvironment and inhibit tumor growth in PDAC. (188 words).

Pancreatic ductal adenocarcinoma (PDAC) exhibits aggressive perineural invasion (PNI), a hallmark of poor prognosis observed in 70-100% of cases. Schwann cells (SCs), key components of the tumor microenvironment, drive PNI via multiple pathways, yet the underlying mechanisms remain unclear. This study investigates the hypothesis that PDAC cells and SCs establish a glutamine-glutamate metabolic symbiosis to fuel PNI. Integrated approaches, including LC-MS metabolomics, isotopic tracing, co-culture systems, and in vivo models, were employed to analyze bidirectional metabolite exchange. Molecular assays and functional studies elucidated signaling pathways. The therapeutic potential of targeting glutamine transporters (SLC1A5/SLC7A5) and glutamate receptor NR2A was tested using inhibitors V9302 and PEAQX. SCs secreted glutamine, which PDAC cells internalized via SLC1A5 and converted to glutamate. Glutamate activated SCs through NR2A, inducing ROS/NRF2-expression and upregulating glutamine synthetase (GS) and GLT-1, thereby regenerating glutamine to sustain the metabolic loop. KRAS-ACTN4-p65 signaling amplified this cycle by transcriptionally activating SLC1A5/SLC7A5 and GLS, while leucine uptake via SLC7A5 activated mTORC1 to promote invasion and PNI. In vivo, dual inhibition of SLC1A5/SLC7A5 (V9302) and NR2A (PEAQX) synergistically reduced tumor growth, PNI length, and improved sciatic nerve function in mice. This study identifies a reciprocal glutamine-glutamate metabolic symbiosis between PDAC cells and SCs as a driver of PNI, orchestrated by KRAS-ACTN4-NF-κB signaling and glutamate-NR2A-ROS-NRF2 pathways. Disrupting this axis with V9302 and PEAQX offers a novel therapeutic strategy to target PDAC's metabolic adaptability and neurotrophic microenvironment.

Macrophages constitute a dominant and functionally diverse immune population within the microenvironment of pancreatic ductal adenocarcinoma (PDAC), yet how macrophage heterogeneity contributes to the tumor remains poorly defined. In an institutional cohort of 145 PDAC specimens, we identified a population of multinucleated giant cells (MGCs) of macrophage origin, an entity previously described in chronic inflammation but rarely in cancer. CD68⁺ MGCs were present in 28% of tumors, enriched in squamous, non-glandular regions, and more frequent after neoadjuvant chemotherapy. By integrating spatial transcriptomics and quantitative imaging, we defined the features of these cells, which, compared with MGCs in non-neoplastic inflammatory lesions, lacked canonical polarization markers (HLA-DR, CD163) and displayed a distinctive transcriptional program characterized by upregulation of the POLR2K, TUBA8, COX5B, and VDAC1 genes, which encode proteins involved in DNA repair, oxidative stress, and MYC signaling. Spatial analyses revealed activation of hypoxia and extracellular matrix-remodeling pathways in MGC-associated niches, and experimental hypoxia promoted MGC formation in vitro. Consistent with these data, we found that in the TCGA PAAD dataset a macrophage MGC gene signature was enriched in the squamous PDAC subtype and correlated with poorer overall survival (p = 0.018). Morphometric and immunofluorescence analyses further showed increased 53BP1⁺Ki67⁺ nuclei and nuclear atypia in MGCs, indicating ongoing proliferation despite DNA damage. Together, these data identify MGCs of macrophage origin as an immune cell state shaped by hypoxia and stress signaling, associated with aggressive tumor phenotypes, and potentially exploitable as an immune classifier in PDAC.

The DNA damage response (DDR) is critical for pancreatic ductal adenocarcinoma (PDAC) development and therapeutic responses, including to genotoxic agents. While epigenetic modulators have been shown to contribute to the DDR, how chromatin regulation dictates responses to DNA damage in PDAC remains incompletely understood. Here, we identify Class I histone deacetylases (HDACs) as critical regulators of the DDR. HDAC1/2 direct the genomic distribution of H3K27ac, ensuring sufficient BRD4 and RNA polymerase II (Pol II) occupancy at DDR gene promoters. HDAC inhibition by entinostat shifts the balance of H3K27 acetylation preferentially toward intergenic regions, diverting BRD4 and Pol II from promoters, thereby suppressing DDR gene expression. In line with this, HDAC inhibition heightens DNA damage and sensitizes PDAC to diverse DNA-damaging and DDR-targeting agents. Since the clinical development of HDAC inhibitors has been limited by systemic toxicity, we developed bottlebrush prodrug (BPD) nanoparticles for tumor-selective entinostat delivery. Entinostat-BPD achieved tumor-specific HDAC inhibition while displaying potent efficacy and reduced systemic toxicity. These findings reveal an HDAC-dependent DDR vulnerability and offer combinational and precision targeting strategies to facilitate clinical translation and improve PDAC patient outcomes.

For over a decade the Advanced Prostate Cancer Consensus Conference (APCCC) covers a variety of topics that greatly impact daily practice. In 2025, a dedicated event was organised to discuss key questions in clinical management of patients with prostate cancer (PC) related to diagnostic tools (APCCC Diagnostics). Here we present the voting results of the APCCC Diagnostics questions. OBJECTIVE; DESIGN, SETTING, AND PARTECIPANTS: APCCC Diagnostics 2025 is a pilot project. The scientific committee for APCCC Diagnostics 2025 developed 88 multiple-choice consensus questions on six different topics. Prior to the conference, the panel members (''panellists'') voted on these questions via a web-based survey. Consensus was defined as ≥75% agreement, with strong consensus defined as ≥90% agreement. Consensus was only reached on 17 of 88 questions (19%), of which six (7%) received a strong consensus. Specifically, consensus was reached for two of 17 questions (14%) in "how to diagnose PC"; seven of 16 (44%) in "how to stage PC"; three of 14 (21%) in "Biochemical Recurrence Scenario"; two of 11 (18%) in "metastatic disease: what to do?"; zero of 18 (0%) in "monitoring metastatic PC"; and three of 12 (25%) in "radioligand therapy and imaging." The voting results and their discussion may assist physicians in navigating controversial areas of clinical management related to diagnosis, staging, and restaging in the different clinical settings for PC, particularly where high-level evidence is scarce or conflicting. The findings can also help funders and policymakers in prioritising areas for future research.

CDK12 mutations occur in 2-7% of metastatic prostate cancers (mPCa) and are considered to be exclusively somatic. Here, we identified five patients with mPCa (ages 44-62) harboring germline CDK12 truncating variants among 4,535 tested (0.1%). All had CDK12-driven cancers defined by an additional somatic CDK12 variant and the CDK12-specific hallmark genomic instability signature characterized by hundreds of tandem duplications. Two patients had multiple independent CDK12-driven tumors with distinct secondary somatic CDK12 variants. Germline CDK12 truncating variants were enriched in mPCa compared to gnomAD V4.1.0 controls (n=807,162; odds ratio 11.4, 95% CI 3.6-27.8) and V2.1.1 non-cancer controls (n=134,187; odds ratio 29.6; 95% CI 6.8-28.6). Family history revealed multiple related individuals with prostate or ovarian cancer, and germline variant inheritance was confirmed in the two tested pedigrees. Our data suggest that germline CDK12 truncating variants are a rare driver of lethal mPCa.

Local and androgen-deprivation therapies are guideline recommendations in localized and metastatic prostate cancer, respectively. However, in select patients the alternative use of radiopharmaceutical therapies may be considered to delay or avoid quality-of-life decline associated with other systemic treatments.

Multiparametric magnetic resonance imaging (mpMRI) is the reference modality for detecting clinically significant prostate cancer (csPC). Biparametric MRI (bpMRI), which omits contrast, has emerged as a streamlined alternative. Recent studies have demonstrated the noninferiority of bpMRI to mpMRI for csPC detection in biopsy-naïve men. We performed an updated systematic review and meta-analysis of head-to-head studies comparing bpMRI and mpMRI for csPC detection, and conducted a noninferiority analysis. Literature databases were searched up to September 2025 for head-to-head studies. Eligible studies enrolled men with suspected PC and used biopsy or prostatectomy as the reference standard. The primary outcome was diagnostic accuracy for csPC at per-patient and per-lesion levels; detection of any PC was a secondary outcome. Noninferiority was assessed using a margin of -5% applied to paired absolute differences (bpMRI - mpMRI) in sensitivity and specificity via a random-effects model. A total of 40 studies (9403 patients) were analyzed. For csPC, the paired absolute differences were -2.3% (95% confidence interval [CI] -4.1% to -0.5%) for sensitivity, and +1.8% (95% CI -0.4% to +4.0%) for specificity, which confirm noninferiority at the patient level. At the lesion level, bpMRI was noninferior for specificity but not sensitivity, probably because of fewer studies and greater heterogeneity. bpMRI is noninferior to mpMRI for csPC detection at the patient level. At the per-lesion level, noninferiority was demonstrated for specificity but not for sensitivity. Broader implementation should occur in settings with assured image quality, and further work is needed to define minimum quality-control standards required for adoption.

Prostate-specific membrane antigen (PSMA)-targeted PET imaging plays a well-established role in staging, detecting recurrence, and selecting patients for PSMA-targeted radiopharmaceutical therapy in prostate cancer. We present a case of PSMA-expressing recurrent prostate cancer in the pelvis, involving the urinary bladder wall and pelvic lymph nodes that was not completely visualized on 18 F-DCFPyL PET/CT, but clearly delineated on 18 F-rhPSMA-7.3 PET/CT. This discrepancy is attributed to the significantly lower urinary excretion and consequent low urinary bladder activity with 18 F-rhPSMA, yielding high target-to-background ratios in the pelvis, and highlighting its potential diagnostic advantage in evaluating pelvic lesions near the urinary bladder.

We report a case involving a 66-year-old man with prostate adenocarcinoma, referred for initial staging using 99m Tc-PSMA and 99m Tc-MDP bone scan. The whole body PSMA scan revealed foci of increased tracer uptake in the right knee and the surrounding periarticular space above the knee. Subsequent 99m Tc-PSMA SPECT/CT imaging demonstrated evidence of arthritis characterized by effusion in the periarticular region, connected to the joint in the right knee. Notably, the patient had received an intra-articular corticosteroid injection just 2 days before the scan. This case highlights the necessity of recognizing atypical 99m Tc-PSMA uptake patterns to differentiate from metastasis, thereby enhancing diagnostic accuracy.

The prostate-specific membrane antigen (PSMA)-targeted radioligand [177Lu]Lu-PSMA-617 is an approved treatment for metastatic castration-resistant prostate cancer (mCRPC). However, identification of genomic biomarkers associated with treatment efficacy remains a critical unmet need. We used a customized 78-gene panel to analyse circulating tumour DNA (ctDNA) from 150 patients with mCRPC included in a prospective [177Lu]Lu-PSMA-617 registry. Undetectable ctDNA was a significant and positive prognostic biomarker at both baseline (before treatment) and at week 6 (before cycle 2 of [177Lu]Lu-PSMA-617). Quantification of the baseline ctDNA fraction enhanced prognostic stratification irrespective of high or low PSMA expression on position emission tomography imaging. Undetectable ctDNA at week 6 was linked to a superior treatment benefit independent of prostate-specific antigen response. FOLH1 alterations were identified as a potential novel therapeutic resistance mechanism. Our data highlight the potential utility of ctDNA in optimising patient selection, improving therapeutic monitoring, and dissecting genomic mechanisms of resistance to [177Lu]Lu-PSMA-617, and further prospective validation is warranted.

Tumors remodel extracellular matrix (ECM) and glycosylation, yielding epitopes with restricted or limited detectability in normal adult tissues. Here, we evaluated the O-glycosylated IIICS domain of fibronectin (Tn-FN) as a chimeric antigen receptor (CAR) T cell target. FDC6-BBζ CAR T cells recognizing Tn-FN were benchmarked against EDB-FN-targeted L19-BBζ and Tn-MUC1-targeted 5E5-BBζ. FDC6-BBζ mediated robust, antigen-dependent activation and cytotoxicity, outperforming L19-BBζ and matching 5E5-BBζ in vitro and in NSG xenografts of prostate cancer. FDC6-BBζ and 5E5-BBζ CAR T cells achieved durable tumor control with increased intratumoral CD3⁺ infiltration and reduced tumor-collagen overlap. Cytotoxicity required intact tumor interferon-γ (IFNγ) receptor 1; L19-BBζ further depended on Fas, whereas FDC6-BBζ and 5E5-BBζ were less Fas-dependent. Tumoral toll-like receptor (TLR) 2/6 or TLR4 agonism restored FDC6-BBζ killing of IFNγR1-deficient targets and induced broad inflammatory and stress-response programs. Pharmacologic perturbation implicated caspase-dependent mechanisms and a contribution from inflammasome-linked signaling, whereas ferroptosis blockade did not abrogate restored killing. These findings establish Tn-FN as a glycoform-restricted, ECM-derived CAR target and show that innate agonists can reprogram tumor state to overcome resistance from impaired IFNγ signaling.

Metastatic castration-resistant prostate cancer (mCRPC) remains an aggressive disease with limited response to systemic therapies despite androgen deprivation. Immune-excluded prostate tumors are typically resistant to immunotherapy. STAR0602 is a selective bifunctional T-cell agonist composed of an antibody targeting Vβ6 and Vβ10 T-cell receptor chains fused to human interleukin-2 that selectively expands Vβ6+ CD8+ memory T cells and has demonstrated clinical activity as a monotherapy in anti-programmed death-ligand 1-resistant tumors (NCT05592626). We hypothesized that combining poly ADP-ribose polymerase (PARP) inhibition with Vβ-directed T-cell activation would enhance antitumor immunity and promote polyclonal T-cell responses in immune-excluded prostate cancer. The antitumor activity of the PARP inhibitor olaparib combined with mSTAR1302, the murine surrogate of STAR0602, was evaluated in TRAMP-C2 and RM-1 prostate tumor models. Tumor growth, survival, and immune responses were assessed by flow cytometry and functional depletion studies. The role of tumor-intrinsic TRAIL-R2 signaling was evaluated using a TRAIL-R2 knockout TRAMP-C2 model, and clinical relevance was explored by assessing TRAIL-R2 expression in tumor samples from patients treated with olaparib in a Phase 2 clinical trial (NCT02484404). Combination therapy with olaparib and mSTAR1302 induced significant tumor regression and improved survival compared with either agent alone. Treatment increased tumor-infiltrating lymphocytes, expanded activated Vβ13+ CD4+ and Vβ13+ CD8+ T cells, reduced immunosuppressive populations, and enriched stem-like progenitor exhausted CD8+ T cells. Depletion studies demonstrated that Vβ13+ CD4+ T cells, Vβ13+ CD8+ T cells, natural killer cells, and interferon-γ were required for therapeutic efficacy. TRAIL-R2 knockout experiments demonstrated that tumor-intrinsic TRAIL-R2 signaling contributes to the antitumor activity of the combination. Mechanistically, these findings support a model in which PARP inhibition sensitizes tumors while mSTAR1302-mediated Vβ13+ T-cell expansion initiates a broader polyclonal antitumor immune response associated with antigen spreading and recognition of multiple tumor antigens and neoepitopes. These findings suggest that combining tumor-sensitizing therapies with selective T-cell activation may represent a broader strategy to overcome immune exclusion in solid tumors. Together, these results provide mechanistic rationale for clinical evaluation of olaparib in combination with STAR0602 in patients with mCRPC who have progressed on androgen deprivation therapy.

Prostate cancer, a prevalent malignancy in the male reproductive system, poses significant therapeutic challenges due to the development of resistance to androgen deprivation therapies such as bicalutamide. While current research predominantly focuses on androgen receptor (AR)-dependent mechanisms of resistance, non-AR-dependent pathways remain poorly understood. Here, we report a novel non-AR-dependent mechanism of bicalutamide resistance centered on DUOXA1, a maturation factor for dual oxidases (DUOX) that catalyzes hydrogen peroxide (H2O2) production. Our analysis of RNA sequencing data from bicalutamide-resistant and sensitive prostate cancer cells revealed DUOXA1 as a significantly downregulated gene in resistant cells. We demonstrate that hypoxia in the prostate cancer microenvironment enhances HIF1α transcriptional activity, leading to increased DNMT1 expression. DNMT1, an epigenetic modifier, mediates the methylation of the DUOXA1 promoter, thereby silencing its expression. This epigenetic silencing of DUOXA1 inhibits ferroptosis, a form of regulated cell death characterized by iron metabolism disruption and lipid peroxidation, thereby promoting bicalutamide resistance. Our findings indicate that DUOXA1 can enhance bicalutamide resistance by promoting ferroptosis through ROS generation. This study not only provides mechanistic insights into the role of DUOXA1 in bicalutamide resistance but also highlights the HIF1α-DNMT1-DUOXA1 axis as a critical regulator of resistance. Our work suggests potential therapeutic strategies to overcome resistance through epigenetic modulation and activation of DUOXA1, offering a novel perspective on the molecular mechanisms of bicalutamide resistance and paving the way for the development of improved treatment approaches for advanced prostate cancer.

Prostate cancer (PCa) is the second most frequently diagnosed cancer in men worldwide and the fifth leading cause of cancer-related mortality, presenting urgent unmet clinical needs in diagnosis and treatment. The recognition of the microbiome as a key factor in human health has prompted numerous studies, revealing an exciting new approach to cancer diagnosis and therapy. The bacteriome is the component of the microbiome that is most metabolically active and influenceable by internal and external factors, which is pivotal in the development of cancer. Initial studies exploring the link between the microbiome and PCa focused primarily on the gut bacteriome, which has been implicated in the onset and progression of the disease, as well as in resistance to therapy. Further research has demonstrated that the urologic bacteriome also plays a crucial role in the development of PCa, serving as an important factor for diagnosis and treatment. This review synthesizes current knowledge concerning the significance of the urologic bacteriome in PCa onset, progression, diagnosis/prognosis, and therapy. It also explores the impact of the bacterial metabolome in PCa, emphasizing the importance of this undervalued dimension of the bacteriome. Overall, the review provides a comprehensive analysis of how bacteria and their bioactive metabolites contribute to PCa, highlighting their clinical and therapeutic relevance. It also identifies the existing knowledge gaps, paving the way for the development of new approaches that could enhance PCa diagnosis and treatment.

Taxane-based chemotherapy and immunotherapy are standard treatments for advanced prostate cancer, yet their efficacy is often limited by drug resistance and an immunosuppressive, "cold" tumor microenvironment (TME). To address these challenges, we develop a reactive oxygen species (ROS)-responsive nanoparticle, PTX-Zn NP, for the co-delivery of paclitaxel (PTX) and zinc ions (Zn2+). Within tumor cells, elevated ROS triggers the release of PTX, promoting micronuclei formation and cytosolic double-stranded DNA exposure. Concurrently, Zn2+ amplifies cGAS-STING signaling by enhancing cGAS-DNA binding and inducing mitochondrial damage. In vitro, PTX-Zn NP suppressed tumor cell proliferation, generated ROS and micronuclei, and activated the STING pathway to promote dendritic cell maturation. In vivo, PTX-Zn NP preferentially accumulated in prostate tumors, inhibited tumor growth, and reprogrammed the "cold" TME toward a "hot" phenotype. When combined with anti-PD-L1 therapy, PTX-Zn NP significantly improved antitumor efficacy and promoted long-term immune memory. Overall, this dual-action approach provides a promising strategy to overcome both chemoresistance and immune evasion in advanced prostate cancer.

Genetic heterogeneity contributes to the variable therapeutic responses in cancers. Frequent SPOP mutations and recurrent CHD1 deletions define distinct molecular subtypes of prostate cancer (PCa) with differential responses to anti-androgen therapy. Ferroptosis, an iron-dependent cell death mechanism driven by lipid peroxidation, has emerged as a promising anticancer strategy. Here, we identify SPOP mutations and CHD1 deletion as key genetic determinants of ferroptosis susceptibility in PCa. Using genetically engineered human and murine models, we show that SPOP mutations enhance, whereas CHD1 deletion impairs, the efficacy of ferroptosis inducers targeting GPX4. Mechanistically, SPOP and CHD1 exert opposing effects on ferroptosis by antagonistically regulating the MYC-ACSL4 axis. Furthermore, we demonstrate that targeting cholesterol metabolism with cholesterol-lowering agents restores ACSL4 expression and re-sensitizes SPOP/CHD1 co-deficient tumors to ferroptosis-inducing therapy. Our findings establish SPOP/CHD1 as upstream genetic regulators of ferroptosis and provide biomarker-driven combinatorial strategies to enhance ferroptosis-based therapy in men with advanced PCa.

Circulating nucleic acids are emerging disease markers whose clinical applications are hindered by the lack of rapid, sensitive, convenient, and cost-effective detection assays. Inspired by the natural replication of virus, here, we developed a biomimetic one-step, one-pot, isothermal detection assay named RAPID to realize rapid and sensitive detection of nucleic acids with minimal reliance on instruments. The core element of RAPID is an autocatalytic molecular sensor that exploits the viral replication endonuclease of duck circovirus (i.e., DCV) to transform rolling circle amplification (RCA) from linear to exponential. DCV cleaves target-induced, RCA-generated amplicons into target analogs to prime secondary RCA reactions to catalytically propagate the amplification sensor. RAPID enables rapid (∼10 min), ultrasensitive (attomolar sensitivity), and direct (RNA extraction-free) detection of microRNAs and viral RNAs that is compatible with smartphone-based fluorescence detection devices. RAPID exhibited pronounced clinical translational capability by quantitatively profiling a panel of six miRNAs to achieve accurate discrimination of prostate cancer from benign prostatic hyperplasia that is exceptionally important but challenging in clinics. Furthermore, RAPID demonstrated rapid detection of influenza A viral infections of high accuracy in point-of-care settings. Simple nucleic acid detection assays like RAPID could largely promote the development and application of liquid biopsy molecular diagnostics.

Ferroptosis, an iron-dependent and non-apoptotic regulated cell death mode, has garnered increasing attention in treating cancers with apoptosis resistance. However, the insufficient hydrogen peroxide (H2O2) supply impedes the sustained generation of highly cytotoxic free radicals, resulting in diminished ferroptosis. In addition, monotherapy with ferroptosis inducers frequently shows limited efficacy in cancer management. Thus, promotion of ferroptosis by increasing H2O2 supply, and the development of targeted combination approaches that co-activate multiple cell death pathways are essential to maximize the tumoricidal effects. Herein, we developed a multifunctional targeted nanozyme system pPB-Fe3O4@CaO2-BzATP, in which the pPB targeting cyclic peptide endowed the nanozyme system with enhanced prostate tumor-specific targeting ability. Upon tumor cell internalization, calcium peroxide (CaO2) was hydrolyzed under acidic conditions to produce H2O2 and calcium ions (Ca2+), and iron oxide (Fe3O4) nanozyme catalyzed self-supplied H2O2 to generate highly cytotoxic ·OH, thereby inducing prostate cancer cell death via augmented ferroptosis. Meanwhile, released Ca2+ can trigger calcium overload to elicit apoptosis. Moreover, 2'(3')-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate (BzATP) activated the  P2X7 receptor, which not only enhanced Ca2+ influx to further reinforce calcium overload but also promoted efflux of potassium ions (K+), followed by nucleotide-binding oligomerization domain (NOD)-like receptor pyrin domain containing 3 (NLRP3) inflammasome assembly and ultimately pyroptosis induction. Both in vitro and in vivo studies verified that pPB-Fe3O4@CaO2-BzATP nanozyme system significantly enhanced the tumoricidal efficacy with negligible side effects. Overall, we propose a novel and promising strategy to combat castration-resistant prostate cancer by synergistically integrating ferroptosis, pyroptosis and calcium overload-induced apoptosis into a unified therapeutic modality.

Recurrence after tumor resection and liver transplantation remains a major clinical challenge in hepatocellular carcinoma (HCC). We examined tumor samples from HCC liver transplant patients to identify drivers of disease recurrence. Integration of proteomic profiling of human HCC samples with or without recurrence and T cell killing assays linked the cholesterol esterification enzyme SOAT1 to immune evasion and cancer recurrence. Genetic or pharmacological inhibition of SOAT1 sensitized liver cancer cells to CD8+ T cell-mediated immunosurveillance, anti-programmed cell death 1 (anti-PD-1) therapy, and chimeric antigen receptor (CAR)-T cell therapy and increased intratumoral CD8+ T cell infiltration. Mechanistically, the inhibition of SOAT1-mediated cholesterol esterification disrupted intratumoral cholesterol and lipid metabolism and reduced unsaturated fatty acids and prostaglandin E2 production; this impaired the antioxidant capacity and metabolic resilience of cancer cells under immune attack. Accordingly, SOAT1 inhibition enhanced immunotherapy efficacy in obesity-associated tumors and under immunosuppressant-induced conditions. Thus, cholesterol esterification supports redox fitness and cancer cell resilience upon immune attack, suggesting a strategy to prevent recurrence and improve immunotherapy outcomes.

Hepatocellular carcinoma (HCC) surveillance in patients with cirrhosis remains suboptimal, with inadequate early-stage detection. α-Fetoprotein (AFP) demonstrates insufficient sensitivity. Circulating methylated septin 9 (SEPT9) has shown diagnostic promise. To determine whether methylated SEPT9 improves detection of HCC when combined with AFP in patients with cirrhosis undergoing surveillance. In this prospective, cross-sectional, diagnostic accuracy study, patients with cirrhosis undergoing routine HCC surveillance with ultrasonography and AFP were enrolled at 2 French academic centers from February 2018 through October 2024. HCC was diagnosed per international guidelines with centralized radiologic review, blinded to methylated SEPT9 results. Data were analyzed from October 2025 to January 2026. Plasma methylated SEPT9 was analyzed from 3 independent plasma aliquots and classified by number of positive replicates (single-positive, double-positive, or triple-positive). Serum AFP was evaluated at a threshold of 20 ng/mL. Biomarkers were evaluated individually and in combination using disjunction logic (tier 1; maximizing sensitivity) or conjunction logic (tier 2; maximizing specificity). The primary outcome was the presence of HCC at enrollment. The primary end point was comparison of the area under the receiver operating characteristic curve (AUROC) between methylated SEPT9 and AFP. Secondary end points included diagnostic performance stratified by Barcelona Clinic Liver Cancer (BCLC) stage. Among 574 participants, 414 (72.1%) were male, and the median (IQR) age was 63 (57-70) years. A total of 118 had HCC, including 51 (43.2%) with BCLC stage 0-A. Methylated SEPT9 outperformed AFP (AUROC: 0.79 [95% CI, 0.74-0.84] vs 0.71 [95% CI, 0.66-0.76], respectively; P = .002; posterior probability of superiority >99.8%). Tier 1a (at least single-positive methylated SEPT9 or AFP >20 ng/mL) achieved 87.8% (95% CI, 81.6-93.5) sensitivity and a negative likelihood ratio of 0.2 (95% CI, 0.1-0.3). Among 64 HCC cases missed by AFP, tier 1a recovered 50 (78%). For BCLC 0-A disease, tier 1a sensitivity was 74.5% (95% CI, 62.2-86.5) vs 23.5% (95% CI, 12.5-35.8) for AFP, 3.2-fold increase. Tier 2 (triple-positive methylated SEPT9 and AFP >20 ng/mL) achieved 99.6% (95% CI, 98.9-100) specificity, a positive likelihood ratio of 76.0 (95% CI, 26.9-181.0), and a diagnostic odds ratio of 112.1 (95% CI, 37.8-294.7). In this diagnostic study, combining methylated SEPT9 with AFP substantially improved HCC detection in patients with cirrhosis, particularly for early-stage disease amenable to curative treatment. Prospective studies are needed to determine whether improved detection translates into survival benefit.

The Barcelona Clinic Liver Cancer (BCLC) classification has been the mainstay for prognostic assessment and initial treatment selection in hepatocellular carcinoma (HCC) for more than two decades. It is widely clinically accepted and has been reaffirmed in the recently updated European Association for the Study of the Liver (EASL) Clinical Practice Guidelines on the management of HCC. Its design is based on simple clinical and imaging parameters, which makes it highly applicable in clinical practice. However, it does not fully utilise all the information potentially encoded in routine radiology imaging. With artificial intelligence (AI) methods now maturing, we have a robust fully automated way to extract and quantify digital imaging features without much user input and with high precision. Therefore, AI could bridge quantitative imaging into clinical decision-making, together with the existing BCLC classification. However, despite substantial AI advancements in many fields such as automated tumour volumetry, radiomics, detection of metastatic lesions, and the identification of opportunistic imaging biomarkers, a translational gap persists. While challenges related to technical, administrative, cost, and training factors have to be taken into account, a certain aversion to change, as well as an absence of standardised AI validation and missing workflow integration hamper clinical implementation in routine care. This article aims to evaluate current AI-quantified imaging parameters and their potential for synergy with the established BCLC classification.

More than 100 million individuals in rural areas of China are suffered from Fatty Liver Disease (FLD). However, health clinics in remote regions often lack the necessary professional expertise and expensive ultrasound equipment for regular liver disease screening. Delayed treatment frequently leads to liver cirrhosis and cancer, imposing substantial economic burden on both public health systems and affected families. Traditional Chinese Medicine emphasizes the strong association between tongue characteristics and liver health. Leveraging machine learning to model the relationship between tongue images and FLD can enable rapid, non-invasive, large-scale screening in medically underserved areas. However, existing studies in this domain often rely on small-scale private datasets, which can result in unverifiable model performance. Moreover, most studies have employed generic convolutional neural networks for feature extraction, causing a lack of interpretability. The goal of our research is to address above-mentioned questions. In this study, we first introduced a Multi-source Feature Fusion-based Tongue Diagnosis Framework for FLD diagnosis (MFF-TDF). In addition, we developed and released a standardized tongue image dataset with physiological indicators and FLD annotations, comprising 5,717 samples, which to our knowledge is the largest public dataset in this domain. Finally, we evaluated the effectiveness of the proposed method through extensive experiments and enhanced model interpretability using shapley additive explanations and counterfactual analysis. When conducting fusion modeling with tongue images and some basic physiological indicators (such as sex, age, height, etc.), FLD's prediction performance in the population reached F1-score 0.797, Recall 0.847, and AUC 0.924. This performance significantly exceeds that of the state-of-the-art methods published in this domain. This study developed an automated and explainable method for tongue diagnosis that facilitated the low-cost, speedy screening of FLD in large-scale populations, and contributed the largest public dataset to support future modeling research in this field.

Targeting transcriptional condensates is an emerging paradigm for cancer therapy. A key player is the transcriptional coactivator YAP (Yes-associated protein), which drives tumor-specific programs that fuel tumor progression and therapeutic resistance. Cyclin K, partnered with cyclin-dependent kinases (CDKs) CDK12/CDK13, is essential for transcription elongation, but its role in specific oncogenic programs was unclear. Here, we identify Cyclin K as an essential vulnerability across multiple cancer types. The CDK12/Cyclin K complex binds YAP via Cyclin K and forms a regulatory condensate to bridge YAP phosphorylation by CDK12. Such a phosphorylation at threonine-398 impedes YAP inhibition by its canonical LATS kinases, stabilizes YAP, and enables its further condensation with TEAD4 to stimulate YAP oncogenic activity. Coexpression of CDK12/Cyclin K and YAP predicts sensitivity to Cyclin K inhibitors in hepatocellular carcinoma cells and patient-derived xenografts. Thus, we define CDK12/Cyclin K as a critical regulator of YAP-driven transcriptional addiction and a biomarker for patient stratification who mostly benefit from therapies targeting the CDK12/Cyclin K-YAP axis.

CAR-T cell therapy has shown remarkable success in hematologic malignancies but remains limited in solid tumors such as liver cancer due to antigen heterogeneity, low target antigen density, and an immunosuppressive tumor microenvironment (TME). Cytokine engineering can enhance CAR-T persistence and effector function; however, the optimal cytokine payload may vary depending on tumor type, target antigen expression level, and microenvironmental context, making systematic experimental comparison time-consuming and labor-intensive. Here, we applied a large language model (LLM)-based CAR-T in silico platform to systematically evaluate cytokine engineering strategies, including IL-2, IL-7, IL-12, IL-15, and IL-18, in glypican-3 (GPC3)-targeted CAR-T cells for liver cancer. We used cytokine selection as a biologically grounded benchmark to test whether the platform could recover known CAR-T cell-relevant cytokine biology and support future novel predictions. Computational predictions identified IL-15 as the most effective enhancer, particularly against tumor cells with low GPC3 expression. Guided by these results, we generated cytokine-armored GPC3 CAR-T cells and performed in vitro and in vivo validation. IL-15-engineered CAR-T cells exhibited superior proliferation, persistence, and serial cytotoxicity against GPC3-low liver cancer cells. In human liver cancer xenograft models, IL-15-enhanced CAR-T cells achieved improved tumor control compared with conventional and other cytokine-engineered CAR-T cells. The recovery of IL-15 served as a positive benchmark supporting the validity of the LLM-guided CAR-T in silico workflow. Collectively, this study establishes an LLM-guided framework, schema-constrained for rational cytokine selection in CAR-T engineering and identifies IL-15 as a potent enhancer for targeting antigen-low liver cancers.

The fabrication of self-healing gels typically relies on dynamic covalent bonds, which prove particularly challenging in organic systems. Inorganic subnanowires (SNWs) exhibit excellent structural flexibility and enhanced interactions beyond molecular levels, enabling click-gelation and self-healing of organogels different from polymers and biomaterials. In this study, we synthesize Na-PMo12 SNWs bridged by Na+ and polyoxometalate clusters, which can lock various organic solvents into transparent organogels within 5 s. These organogels exhibit remarkable self-healing capabilities, with visually complete morphological recovery within 20 min even after mechanical disruption. The framework formed by the linkage of Na+ and clusters significantly augments the flexibility of SNWs, and the dense surface ligands of SNWs bring enhanced interactions and entanglements, facilitating the click gelation and self-healing of organogels. The shear-thinning property of the organogel enables its facile injection, making it a promising candidate for applications, such as transarterial chemoembolization (TACE) for liver cancer and injectable drug delivery systems.

A Cr2O3-melamine@MAF-6 derived carbon nanozyme composite (CrN@MDC) was developed and integrated into a lateral flow assay (LFA) for the sensitive and specific quantification of transglutaminase 2 (TGM2), a potential biomarker for liver cancer, in clinical plasma samples. CrN(x)@MDC nanozymes with varying Cr2O3 contents (x = 0, 0.05, 0.10, and 0.15 g) were synthesized, and their peroxidase-like activities were evaluated using a 3,3',5,5'-tetramethylbenzidine (TMB)/H2O2 colorimetric reaction, which identified CrN(0.10)@MDC as the most catalytically active composition. The CrN(0.10)@MDC-based LFA generated a distinct black-to-blue colorimetric signal through nanozyme-catalyzed TMB oxidation, enabling detection of TGM2 concentrations as low as 0.025 nM in both buffer and plasma. This corresponded to an approximately 40-fold improvement in visual detection sensitivity compared with a conventional gold nanoparticle-based LFA. In addition to a low limit of detection (0.0095 nM) in a plasma matrix, the assay exhibited high selectivity against abundant plasma proteins, relevant biomarkers, and liver disease-associated interferents, likely attributable to effective nanozyme surface blocking using bovine serum albumin. Direct analysis of undiluted clinical plasma samples from healthy individuals and liver cancer patients showed that TGM2 concentrations measured using the proposed LFA were consistent with those obtained by a commercial enzyme-linked immunosorbent assay. These results support the applicability of the nanozyme-amplified LFA as a proof-of-concept platform for clinically relevant TGM2 detection.

Oxaliplatin (OXA) serves as a key chemotherapeutic agent in trans-arterial infusion chemotherapy (TAIC) for liver cancer. However, its clinical efficacy is frequently limited by several factors: suboptimal tumor uptake, systemic detoxification mediated by glutathione (GSH), and the activation of cellular DNA repair mechanisms. Herein, we present a hollow MnO2 nanoparticle loaded with OXA, the PEI-HMnO2@OXA, to improve the TAIC effect of OXA. The acidic tumor microenvironment facilitated the release of OXA and triggered PEI-HMnO2 to generate free radicals. When coupled with GSH depletion, this cascade culminated in significant DNA damage. Moreover, the PEI-HMnO2 showed a synergistic effect with OXA by blocking multiple DNA repair genes. On the other hand, by leveraging the enhanced permeability and retention effect of the nano-sized structure, 10-100 times greater tumor uptake and a more pronounced inhibitory effect by TAIC are achieved compared with intravenous or single-drug treatment. Meanwhile, the PEI-HMnO2@OXA enabled real-time MRI monitoring of drug distribution and tumor state, facilitating the treatment guidance. Comprehensive experiments using different cell lines, mouse and rabbit models, and patient-derived HCC OXA-sensitive/resistant organoids were conducted to clarify the tumor-inhibiting effects of PEI-HMnO2@OXA, providing novel insights into cancer management.

Dysregulated cell cycle progression is a hallmark of cancer, promoting uncontrolled cell proliferation. Growth arrest-specific 2 like 3 (GAS2L3), which is highly expressed in hepatocellular carcinoma (HCC) cells, correlates with poor clinical outcomes. However, the role of GAS2L3 in HCC pathogenesis remains unclear. This study aimed to investigate the molecular mechanisms by which GAS2L3 contributes to cytokinesis and hyperproliferation of HCC cells. Loss-of-function assays of GAS2L3 were performed to assess midbody assembly, a process essential for cytokinesis. By analyzing protein structure, we predicted liquid-liquid phase separation (LLPS) potential of GAS2L3, which was validated via fluorescence recovery after photobleaching (FRAP), time-lapse imaging, and in vitro droplet formation assays. Additionally, we generated an intrinsically disordered region (IDR)-deficient model to investigate the role of IDR in LLPS and its effects on cytokinesis, cell cycle progression, and HCC tumorigenicity. Highly expressed GAS2L3 localizes to the midbody and accelerates the cytokinesis of HCC cells by facilitating the recruitment of the charged multivesicular body protein 4B (CHMP4B). Notably, EGFP-GAS2L3 aggregates in the midbody exhibit dynamic behavior. Structural analysis and experimental evidence confirm GAS2L3 undergoes phase separation via its IDR. GAS2L3 scaffolds the CHMP4B condensate formation at midbody through phase separation, thereby accelerating cytokinesis in HCC cells. Moreover, GAS2L3 knockdown or IDR deletion in HCC cells leads to defective cytokinesis, G1 arrest, and significant tumorigenicity inhibition. GAS2L3 critically regulates cytokinesis in HCC through LLPS-mediated remodeling of midbody abscission microenvironment. These findings reveal novel insights into HCC hyperproliferation and suggest GAS2L3 as a potential therapeutic target.

Despite promising development as emerging "off-the-shelf" therapeutics against cancer, natural killer (NK) cells still faced considerable challenges in the solid tumor microenvironment (TME), including poor penetrance and immuno-suppression. Here, we employ spatial and single-cell transcriptomics to reveal a role for Nur77 in NK cell-mediated immunity against hepatocellular carcinoma (HCC). Orthogonal analysis of human and mouse HCC tumors indicate that the expression of NR4A1, encoding Nur77, is associated with NK cell proliferation, activation of the immunostimulatory AP-1 gene regulons, and better disease-free survival in HCC. Conditional ablation of Nr4a1 in NK cells perturbs their homeostasis and accelerates tumor progression in multiple tumor models. Conversely, the agonistic activation of Nur77 in NK cells ex-vivo or in-vivo enhances their anti-tumor functions. Mechanistically, downstream functional assays confirm that Nur77 activation attenuates CD36 expression in NK cells and confers resistance against oxLDL-mediated immunosuppression in the TME. Collectively, our findings highlight the potential of harnessing Nur77 agonism in improving NK cell-based immunotherapy against HCC.

Single-mode biosensor is prone to external interference, and detection of a single biomarker is insufficient for accurate diagnosis of liver cancer. Herein, we report a 7-(Diethylamino)coumarin-3-carboxylic acid (7DCCA) functionalized aptamers induced photoelectrochemical(PEC)/fluorescence(FL) dual-mode aptasensor integrated into a three-channel microfluidic chip for multiplexed detection of three liver cancer biomarkers: alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and Golgi protein 73 (GP73). A robust PEC sensing platform was established using I/S co-doped oxygen-vacancy bismuth oxybromide (ISBO-OVs) modified with gold nanoparticles (Au NPs), with 7DCCA-labeled aptamers (7DCCA-Apt) serving as the unified recognition probes. In the presence of target analytes, 7DCCA-Apts bind to their corresponding targets, dissociating from the complementary strand and releasing into the electrolyte to activate the FL signal; simultaneously, this dissociation reduces the insulating layer thickness and restores the photocurrent response. Furthermore, superoxide anions (•O2-) are generated through the dissolution of oxygen by photogenerated electrons, which reacts with luminol to produce chemiluminescence that as an internal light source to further amplify the PEC signal. The integrated microfluidic aptasensor exhibits a wide linear range (1.0 × 102 fg/mL - 1.0 × 102 ng/mL), low detection limits (31 - 37 fg/mL across the three biomarkers), excellent specificity, stability, and reproducibility for both modes. This strategy enables integrated detection of liver cancer biomarkers, holding great promise for early clinical point-of-care screening.

Chronic liver disease represents a major global public health challenge, and its malignant progression to hepatocellular carcinoma is the leading cause of death among affected patients. Gut microbiota dysbiosis is a critical driver of this process. As the central hub of the "gut-liver axis," the gut microbiota, when disrupted, compromises the integrity of the intestinal mucosal barrier, promoting the translocation of microbial metabolites, such as lipopolysaccharides and aberrant secondary bile acids, to the liver. In turn, key signaling pathways become activated, including TLR4/NF-κB, Wnt/β-catenin, and PI3K/Akt, sustaining persistent hepatic inflammation and oxidative stress. These pathological processes accelerate the progression from liver fibrosis to cirrhosis, promote genomic instability, and suppress tumor suppressor gene expression, paving the way for the malignant transformation of hepatocytes. Leveraging its holistic regulatory properties, characterized by multi-component, multi-target, and multi-pathway actions, Chinese medicine can intervene at multiple stages of this inflammation-to-cancer cascade by modulating both the structure and function of the gut microbiota. It does so first by enriching beneficial short-chain fatty acid-producing bacteria, such as Lactobacillus and members of the phylum Firmicutes, which helps restore the intestinal mucosal barrier, limit endotoxin translocation, and alleviate hepatic inflammation and fibrosis. In parallel, by normalizing bile acid metabolism and reestablishing gut microbial homeostasis, Chinese medicine counteracts the development of a tumor-permissive microenvironment marked by immune suppression and DNA damage in hepatocytes induced by microbial metabolites. At the same time, it enhances anti-tumor immune responses mediated by CD8+ T cells and other immune effectors. Drawing on evidence from multi-omics analyses and clinical studies, this review examines the core mechanisms and recent advances regarding how Chinese medicine monomers and formulations modulate the gut microbiota to impede the progression of chronic liver disease to HCC. It highlights gut microbiota dysbiosis as a key driver of hepatocarcinogenesis and highlights the therapeutic potential of targeted microbiota regulation by Chinese medicine, providing a conceptual foundation and strategic approaches for the precision prevention and treatment of hepatocellular carcinoma.

Secondary myelofibrosis (SMF) represents a late stage of polycythemia vera (PV) and essential thrombocythemia (ET), with overall survival (OS) currently defined by the myelofibrosis secondary to PV and ET prognostic model (MYSEC-PM). To identify additional myeloid neoplasm-associated cancer gene variants (CGVs) associated with SMF outcome, we evaluated next-generation sequencing panel testing in 644 patients within the MYSEC cohort. Overall, 429 (66.6%) patients reported at least 1 CGV, with ASXL1, TET2, and DNMT3A being the most frequently involved. Specific molecular profiles affected OS (P< .001): U2AF1, TP53, or SRSF2 variants (UTS; 9.3%; median OS, 4.1 years) and ASXL1 without UTS (25.3%; median OS, 8.4 years). By integrating these genetic signatures within the MYSEC-PM through penalized Cox regressions, we identified the following independent predictors (P< .0001 to .02): hemoglobin level <11 g/dL (1 point), circulating blasts ≥3% (2 points), platelet count <150 × 109/L (2 points), age (0.21 points/y), ASXL1 without UTS mutations (1 point), and any UTS mutations (3 points). Finally, we developed the MYSEC-molecular prognostic model (MYSEC-mPM) allocating 582 patients with SMF into 4 categories with different OS (P < .001): low (median OS, 18.0 years; score <14), intermediate-1 (8.8. years; score, 14-16), intermediate-2 (4.6 years; score, 17-18), and high risk (1.9 years; score ≥19). Additionally, in 381 patients with SMF and available cytogenetics, the MYSEC-mPM was implemented with complex/monosomal karyotype, generating the karyotype-enhanced MYSEC-kmPM. Our study shows that genomic and cytogenetic profiling improves survival prediction in SMF, outperforming the MYSEC-PM.

Survival has improved substantially for patients with Hodgkin lymphoma (HL), but long-term quality of life (QoL) remains incompletely understood. This was a Danish, nationwide, cross-sectional study of QoL among persons with a diagnosis of HL matched 1:10 to general population comparators. Questionnaires included the HeartQoL, the European Organization for Research and Treatment of Cancer Quality of Life Core-30 (QLQ-C30), the Short Form-36 (SF-36), and the EuroQoL Health Questionnaire (EQ-5D). Mean differences (MD) were estimated using linear regression adjusted for sex and age, and stratified by time since diagnosis (0-5, > 5-10, and > 10 years). Overall, 1777 patients with HL (42% of 4156 invited) and 6166 matched comparators (14% of 41 558 invited) responded, and median age was similar (HL: 59, comparators: 61). Most had classical HL (92%). HL groups had consistently and significantly lower QoL than their respective comparators, with 0-5, > 5-10, and > 10 years post-diagnosis MDs of -0.27, -0.28, and -0.24 for the HeartQoL, -7.4, -7.6, and -5.6 points for the QLQ-C30 summary score, -4.5, -4.9, and -4.2 points for the SF-36 physical component summary, and -0.05, -0.05, and -0.04 for the EQ-5D index. The relative difference between the HL group and comparators decreased from baseline to > 10 years post-diagnosis, but differences remained clinically important. The most pronounced symptoms were fatigue and dyspnea. To summarize, persons with HL experience reductions in QoL compared with the general population, even > 10 years post-diagnosis. The observed differences were clinically relevant within several domains and emphasize the need for a multidisciplinary approach to survivorship care.

The theory of immunosurveillance posits that T cells eliminate clones harboring nonself-antigens generated by somatic mutations. Although a role of immunosurveillance in cancer is supported by evidence, whether it affects precancerous expansions has not been well established. Here we studied the association between MHC-variant binding and risk of clonal hematopoiesis (CH), a blood precancer state, predicting MHC binding affinity toward CH hotspot variants in 380,000 UK Biobank participants and examining the relationship between predicted binding to each variant and its expansion risk. Despite the study being powered to detect subtle differences in selective pressure, we did not find associations between predicted binding and CH prevalence for any of the examined variants. In CH-affected individuals, we identified no relationship between predicted variant binding and clone size. Overall, we found no evidence that the MHC genotype affects which variants expand in CH, suggesting a limited role for immunosurveillance in shaping clonal expansions in the blood.

Targeted immunotherapies have revolutionized outcomes for lymphoid malignancies, but success in myeloid neoplasms is limited by the lack of amenable targets and immunologically hostile tumor microenvironment (TME). Myeloproliferative neoplasms (MPNs) are chronic myeloid blood cancers, a third of which are driven by mutations in calreticulin. Calreticulin mutant protein (mutCALR) binds and activates thrombopoietin receptor (TpoR) and results in the display of the mutCALR-TpoR complex on the extracellular membrane of disease-driving cells, thus exposing a therapeutic vulnerability. Here, we present a chimeric antigen receptor T cell (CAR T cell) therapy that specifically targets mutCALR+ cells, both in vitro and in vivo. The CAR T cell therapy selectively depleted mutCALR+ stem cells in samples from patients with myelofibrosis without affecting healthy stem cells and improved survival in mutCALR leukemia xenograft models. To mimic myelofibrotic marrow, we developed a human "chimeroid" model and showed no decrease in the potency of CAR T cell-mediated target cell killing even in a fibrotic TME. We also devised a method to boost the cell surface expression of mutCALR in CD34+ cells isolated from patients with accelerated/blast phase MPNs (defined as >10% blasts in the peripheral blood or bone marrow), enhancing CAR T cell targeting. This study presents a therapeutic strategy with potential to eradicate mutCALR-driven malignancies and highlights an innovative strategy to evaluate blood cancer-targeting immunotherapies in a relevant TME.

Chemotherapy resistance remains a critical challenge in the treatment of patients with cancer, including acute myeloid leukemia (AML). Although genetic alterations can contribute to resistance, the role of rapid-adaptive nongenetic mechanisms, particularly transcription dynamics, remains poorly understood. In this article, we demonstrate that short-term treatment of AML cells with the widely used chemotherapeutic cytarabine (AraC) leads to the rapid emergence of a cell population with significant RNA induction and increased AraC resistance in cell lines and primary patient samples. Mechanistically, transcriptomic and targeted high-resolution analysis of transcription dynamics using single-molecule RNA FISH revealed rapid induction of transcriptional dynamics and upregulation of key transcription factors (TF)-which we term "AraC rapid response TFs." Functionally, short-term pre- and cotreatment with RNA transcription inhibitors suppressed chemotherapy-induced RNA induction and prevented resistance acquisition in vitro and in vivo. Furthermore, CRISPR-mediated suppression of TFs PU.1 and GATA1 significantly attenuated AraC resistance. Our findings reveal a role of rapid-adaptive transcriptional dynamics in AML chemotherapy resistance. This study reveals a role of rapid-adaptive transcriptional dynamics in AML chemotherapy resistance, highlighting master TFs as key regulators. These insights offer a pharmacologically accessible approach to potentially alleviate the major clinical problem of chemotherapy resistance.

Benzene is a well-established environmental leukemogen, but how benzene-induced myelosuppression evolves into rapid malignant transformation remains unclear. To deconstruct this progression, Mll-Af9 chimeric mice were subjected to chronic benzene inhalation. Following exposure, mice exhibited prolonged hematotoxicity, but the initially suppressed white blood cells and CD45.2⁺ pre-leukemic cells progressively rebounded, significantly exceeding control levels by week 10. Serial colony-forming assays revealed suppressed clonogenic capacity at week 8, followed by a robust enhancement at week 10 that was predominantly driven by sustained colony-forming unit-granulocyte-macrophage progenitor (CFU-GM) expansion. These phenotypic shifts pinpointed the 8-10-week interval as the critical tipping point for the suppression-to-proliferation switch. Single-cell RNA sequencing identified granulocyte-macrophage progenitors (GMPs) as the principal cellular drivers of this transformation. During the suppressive phase, a distinct GMP subset displayed cell cycle arrest yet aberrantly co-activated myeloid differentiation (CEBPA) and pro-leukemic self-renewal (HOXA9/MEIS1) programs, accompanied by marked S100a8/S100a9 upregulation. This stress-adapted phenotype transcriptional state was associated with subsequent transcriptional reversal and clonal outgrowth that ultimately shortened overall survival. Clinical relevance was further supported by The Cancer Genome Atlas cohort, where elevated S100A8/S100A9 expression correlated with poorer survival in acute myeloid leukemia patients. Collectively, these findings delineate a stress-driven evolutionary pathway linking benzene-induced marrow suppression to early pre-leukemic adaptation and highlight S100a8/S100a9-associated transcriptional programs as potential early molecular features of benzene-related leukemogenic progression.

Relapsed/refractory B-cell acute lymphoblastic leukemia (B-ALL) remains a leading cause of cancer-related death in children and young adults. While CD19-directed chimeric antigen receptor T cell (CAR-T) therapy offers promise, high rates of long-term failure underscore the need to understand resistance mechanisms. Our studies found p53 inactivation promotes CAR-T resistance in human pre-B-ALL cell lines. Through genome-wide CRISPR/Cas9 screening of CAR-sensitive TP53-wildtype and CAR-resistant TP53-mutated CD19 + B-ALL cell lines, we found the Fatty Acid Transport Protein 2 (FATP2, encoded by SLC27A2) is a leukemia-intrinsic mechanism of CAR-T resistance in TP53-mutant B-ALL. High SLC27A2 expression in pediatric B-ALL patients correlate with worse survival outcomes following conventional chemotherapy. Using B-ALL cell lines and patient-derived xenografts, we show that FATP2-expressing TP53-mutant B-ALL resistance to CAR-T is dependent on exogenous lipid uptake to fuel fatty acid oxidation (FAO) and cell survival, which can be pharmacologically targeted through inhibition of neutral lipolysis and CPT1. These findings identify FATP2-mediated fatty acid uptake and downstream FAO as a potential target to improve existing CAR-T efficacy in human B-ALL.

Cancer cells adapt to treatment, leading to the emergence of clones that are more aggressive and resistant to anti-cancer therapies. We have a limited understanding of resistance mechanisms as we lack technologies to map cancer evolution under the selective pressure of treatment. To address this, we present a hierarchical, dynamic lineage-tracing method, FLARE (Following Lineage Adaptation and Resistance Evolution). We use FLARE to track the progression of acute myeloid leukemia (AML) cell lines treated with Cytarabine (AraC), a front-line treatment in AML, both in vitro and in vivo. We map distinct cellular lineages in both murine and human AML cell lines that are predisposed to AraC resistance. Using FLARE, we identify treatment-naïve populations responsible for seeding resistance that are characterized by upregulation of stemness markers and a cell adhesion-associated AraC-resistant lineage signature. We find that expression of this signature in pediatric AML is associated with the expansion of HSC-like malignant cells at relapse and significantly shorter overall survival. These findings underscore the role of pre-existing lineage states in driving relapse and establish FLARE as a platform for uncovering the evolving, heritable transcriptional programs that underlie tumor evolution.

Primary cutaneous T-cell lymphomas (CTCL) are a heterogeneous group of extranodal non-Hodgkin lymphomas. Outcomes for patients with advanced-stage disease are suboptimal, as few complete and durable responses are achieved with currently available therapeutic agents. However, improved understanding of oncogenic transcriptional programs in malignant T cells and their engagement with the tumor microenvironment (TME) may unveil therapeutic vulnerabilities. Therefore, we have compiled the largest available scRNA-seq CTCL atlas that includes >2 million skin and blood cells from 116 individual patients. We identified recurrent transcriptional programs in malignant T cells, a subset of which are associated with GATA-3 dependent transcriptional programs, especially in the setting of large cell transformation and advanced-stage disease. Many of the transcriptional programs identified are therapeutically targetable with clinically available agents, including HDAC, XPO1, CDK9, JAK/CSF1R, and IKZF1/IKZF3 antagonists. The CTCL TME is dominated by infiltrating and exhausted effector and cytotoxic T cells that are restrained by a robust infiltrate of regulatory T cells, transcriptionally polarized monocytes/macrophages, and cancer-associated fibroblasts. Collectively, these findings have significant implications for the rationale design of combinatorial strategies targeting immune checkpoints, including PD-1. We have identified transcriptional programs, driven by oncogenic transcription factors, and constituents of the TME as therapeutic vulnerabilities in CTCL, and hope that the CTCL atlas constructed will provide a valuable resource for future studies exploiting these therapeutic vulnerabilities.

Acute myeloid leukemia (AML) is a blood cancer with poor survival outcomes. Acute respiratory failure frequently occurs due to leukemia infiltration of the lungs. Underlying mechanisms remain unexplored and therapeutic interventions remain empiric. Here we map the AML lung microenvironment at spatial and single-cell resolution. We show that extensive remodeling is coupled with inflammation and impaired tissue integrity and function. Steroid treatment significantly reduces AML burden and lung infiltration, improving oxygenation and pulmonary function. As a mechanistic correlate, the S-type lectin Lgals9 is triggered by inflammation and mediates cell-cell interactions within infiltrated lungs. Also, the alarmin IL-33 and its receptor (IL-1RL1) are involved in cell-cell interactions within the leukemic lung microenvironment. Targeting either the Lgals9 or IL-33 axis significantly decreases overall AML burden and lung infiltration through effects on both the immune microenvironment and AML cells. Our studies delineate pulmonary infiltration phenotypes in acute leukemia, enabling new treatment strategies.

The unfolded protein response (UPR) is a stress-adaptation pathway and therapeutic target in cancer, yet its pro-survival versus pro-death outcome is difficult to predict because the three ER sensors, PERK, IRE1α, and ATF6, are highly interconnected. Transcriptomic analyses identified sensor-specific gene signatures associated with patient survival across malignancies, and indicated that low IRE1α activity (low XBP1 signature or higher expression of RIDD targets) correlates with improved outcome. We developed SNUPR (single nuclei analysis of the unfolded protein response), an accessible flow cytometry approach that profiles all three branches in nuclear suspensions. SNUPR reveals marked heterogeneity of UPR activation across cancer cell lines that cannot be inferred from sensor expression. This heterogeneity is derived from differences in the strength and duration of PERK-mediated translational inhibition, which gates downstream translation-dependent IRE1α and ATF6 transcriptional programs. Finally, in multiple myeloma, we show that bortezomib-tolerant cells depend on IRE1α activity for survival, linking UPR state to proteasome-inhibitor resistance and positioning SNUPR to guide branch-selective targeting.

Chimeric antigen receptor (CAR)-T cells represent a recent clinically validated modality in cancer therapy and beyond. However, broad industrial implementation faces technological, logistical, regulatory and financial challenges. A major bottleneck is the ex vivo production process, where cytokines are essential and product-determining constituents. To better understand the complexity of cytokine utilization throughout the production process, we rigorously reviewed the existing literature, including extended manufacturing parameters from 292 available clinical reports. We found a progressive reduction of interleukin-2 exposure, driven by its association with unfavourable cell characteristics. Preclinically, this catalysed the evaluation of alternative cytokines with potential to preserve naive phenotypes, support cell expansion and enhance the efficacy of CAR-T cells. Here we illustrate the evolving use of cytokines, reveal non-standardized clinical CAR-T cell manufacturing parameters, and summarize preclinical and next-generation concepts, which may improve manufacturing efficiency, cost-effectiveness and therapeutic outcomes. Our observations may further guide the development of cytokine-armouring strategies to promote in-patient expansion and persistence, even as innovations such as shortened manufacturing and in vivo engineering techniques could reduce reliance on cytokines during ex vivo culture.

Vγ9Vδ2-T cells form a conserved T-cell subset known for its potent intrinsic antitumor activity and versatility in recognizing diverse cancer types independently of the major histocompatibility complex. Previously, we reported the preclinical activity of a bispecific T-cell engager (bsTCE) specific for both CD1d and the Vδ2-TCR that engaged both Vγ9Vδ2-T and type 1 natural killer T cells to CD1d-expressing hematological malignancies, including multiple myeloma (MM) and acute myeloid leukemia (AML). Here, we evaluated whether various standard-of-care drugs for patients with MM and AML/myelodysplastic syndromes (MDS) affected the in vitro antitumor activity of CD1d-Vδ2 bsTCE-activated Vγ9Vδ2-T cells. MM and AML/MDS standard-of-care drugs were tested for antagonistic, additive, or synergistic effects on CD1d-Vδ2 bsTCE-induced and Vγ9Vδ2-T cell-mediated tumor cell lysis. Based on observed synergy, the immunomodulatory drug (IMiD) lenalidomide was studied in more detail, exploring effects on Vγ9Vδ2-T cell proliferation, phenotype, cytokine profile, and cytolytic activity, as well as its mechanism of action, using healthy donor peripheral blood mononuclear cells (PBMC) and MDS patient PBMC and bone marrow samples. Lenalidomide exerted synergistic activity on CD1d-Vδ2 bsTCE-triggered Vγ9Vδ2-T cell antitumor activity and was found to substantially enhance CD1d-Vδ2 bsTCE-induced Vγ9Vδ2-T cell activation, degranulation, Th1-type cytokine secretion, and expansion. Stimulatory effects of lenalidomide on Vγ9Vδ2-T cell effector functions were mediated via enhanced intracellular CD28 phosphorylation, activation of Notch signaling, and interleukin-2 release. Importantly, lenalidomide facilitated otherwise absent proliferation of Vγ9Vδ2-T cells with oncolytic potential in response to CD1d-Vδ2 bsTCE exposure in MDS patient-derived bone marrow samples. Our findings provide a rationale to explore the combination of CD1d-Vδ2 bsTCE and the IMiD lenalidomide in patients with CD1d-expressing hematological malignancies.

A new analysis of tumor data from the MSK-IMPACT dataset revealed that the effects of cancer driver mutations differ based on their context, and that HLA alleles vary considerably by ancestry. Both findings have potential therapeutic implications.

Childhood cancer survival in low-income countries remains below 20%, with late recognition and delayed referral commonly reported. In Northwest Ethiopia, care providers often lack the knowledge, skills, and support systems for recognizing early warning signs of pediatric malignancies. Community caregivers often present only after symptoms become advanced. The University of Gondar Comprehensive Specialized Hospital piloted a quality improvement initiative aimed at achieving earlier diagnoses. This study reports whether a multilevel onsite training and mentorship model could improve early recognition, referral practices, and timely diagnosis of childhood cancer in Northwest Ethiopia. From January 2024 through September 2024, a quasi-experimental pre-post, mixed-methods study design was employed across three tiers of care: primary (e.g., health centers), secondary (general hospitals), and tertiary (specialized hospitals). Eighteen primary-level and 29 secondary-level clinicians completed intensive, on-site training (Ten and Seven days, respectively), while one thousand twenty health extension workers received pictorial outreach modules. A 6-month mentorship program combined monthly onsite visits with remote supervision. Mentees included general practitioners, nurses, health officers, and health extension workers mentored by a multidisciplinary team that included, for example, pediatric haemato-oncologists. Knowledge, attitude, and practice surveys and clinical chart reviews were conducted at baseline and 6 months post-implementation to evaluate patient-journey intervals. These quantitative assessments were integrated with qualitative interviews and focus groups grounded in the consolidated framework for implementation research. The intervention utilized specific curricula focusing on clinical recognition, referral protocols, and documentation. In this study, the participants had a 100% response rate. Median knowledge scores at primary and secondary levels rose from 54.6 (95% CI: 36.4, 63.6) to 90.90 (95% CI: 81.8, 100.0) and from 36.36 (95% CI: 27.3, 45.5) to 90.91 (95% CI: 61.4, 93.7), respectively. Median practice proficiency increased 87.5 (95% CI: 78.7, 100) to 100.00 (95% CI: 100.0, 100.0) at primary level and 68.75 (95% CI: 50.0, 87.5) to 93.8 (95% CI: 86.7, 100.0) at secondary level. The median interval from symptom onset to first health contact fell by 9.3% (from 27.0 days (95% CI: 16.0, 33.3) to 24.5 days (95% CI: 15.0, 32.0)), and the time from first contact to confirmed diagnosis decreased by 54.2%. Treatment initiation interval increased by 11.9%, reflecting ongoing infrastructural constraints. Qualitative findings underscored the roles of supportive leadership, diagnostic supply limitations, cultural beliefs, and referral coordination in shaping outcomes. This quasi-experimental pre-post design without a control group limits strong causal inference, especially in the presence of potential confounders like parallel public-health initiatives and seasonal variations in care-seeking. A context-tailored, multilevel training and mentorship model was associated with improved provider capacity and reduced diagnostic delays in Northwest Ethiopia. While the initiative demonstrated high fidelity and adaptability in conflict-affected settings, achieving timely treatment requires further investment in diagnostic infrastructure. These tools and protocols are well-positioned for national scale-up and integration into routine continuing medical education.

Post-translational modifications (PTMs) dynamically regulate protein function, with metabolite-driven PTMs linking metabolism to protein regulation1,2. We have previously discovered lysine lactylation, showing that lactate can directly modify proteins and influence cancer progression3,4. Recently, pyruvate, another glycolytic metabolite, was shown to directly modify STAT1 at lysine 201, thereby suppressing type I interferon signalling5. Yet, the enzyme governing this modification, its substrate landscape and potential roles beyond innate immunity remain entirely unexplored. Here we report the systematic characterization of lysine pyruvylation (Kpy). Through biochemical and proteomic approaches, we establish the widespread existence of this modification, identifying 88 Kpy sites in mammalian cells. We investigate the dynamic regulation of Kpy upon metabolic perturbations and find that Kpy fluctuates with changes in glycolytic flux and pyruvate levels. Furthermore, we identify sirtuin 3 (SIRT3) as responsible for removing Kpy, while histone acetyltransferase 1 (HAT1) and p300 (EP300) catalyse its addition. Finally, we explore the function of Kpy in transcriptional regulation. Overall, Kpy expands the repertoire of metabolite-driven PTMs and provides insights into how pyruvate directly modulates protein function.

While mutant-specific KRAS inhibitors are approved to treat cancer, a deeper understanding of intratumoral changes driven specifically by KRAS inhibition is needed to maximize therapeutic responses. Here, we used single-cell RNA-seq, flow cytometry, and spatial transcriptomics to distinguish mechanisms of tumor control after KRASG12C inhibition (KRAS(G12C)i) or MEK inhibition (MEKi). Despite both inhibiting the MAPK pathway, KRAS(G12C)i and MEKi drive the adaptation of distinct neoplastic cell fates affecting metabolism and cell cycle regulation, and additive tumor suppression is observed after co-administration. KRAS(G12C)i results in the emergence of a specific, cDC1-driven mature conventional dendritic cell (cDC) state. Co-culture of treated neoplastic cells with cDC1s is sufficient to upregulate maturation markers such as CCR7, intercellular communication analyses suggest activation is augmented through non-immune mediators. Both KRAS(G12C)i and MEKi increase infiltration of cytotoxic T cells, but MEKi, which also targets non-malignant cells, is associated with a reduced capacity for T-cell proliferation and degranulation, consistent with distinct adaptive immune activation mechanisms. We observe that combination treatment of KRAS(G12C)i with anti-PD-1 immunotherapy further expands effector T-cell states, increases clonal persistence, and induces pro-inflammatory macrophages associated with higher overall survival that were largely absent after KRAS(G12C)i alone. Furthermore, combination treatment enhances intercellular communication networks among non-PD-1+ expressing cells that can perpetuate cDC activation. Our findings delineate distinct tumor and immune responses to KRAS and MEK inhibition and identify molecular features of the responding tumor microenvironment that may be leveraged to improve therapeutic efficacy.

Lipid nanoparticles (LNPs), composed of ionizable lipids, phospholipids, cholesterol, and PEGylated lipids, have been successfully used in messenger RNA (mRNA) vaccine development. Despite substantial progress, endosomal entrapment after cellular internalization is still a critical bottleneck limiting the vaccine efficacy. While the efforts to optimize lipid pKa, spatial conformation, and LNP composition have been made, further fine-tuning of these parameters shows diminishing returns in improving effectiveness. Here, we propose a previously unreported parameter, programmable mechanical movement. LNPs are expected to destabilize endosomal membranes through conducting mechanical movements under specific inputs, achieving robust endosomal escape. Specifically, we demonstrate a light-emitting diode (LED)-driven movable lipid (i.e., phenylazothiazole lipid) capable of performing mechanical movements. We integrate the movable lipids into the BNT162b2 formulation from Pfizer-BioNTech. Upon LED irradiation, the movable lipids within LNPs function as molecular rotors, thereby facilitating endosomal membrane destabilization. This strategy has achieved exciting preclinical results in enhancing mRNA-LNP cancer vaccine efficacy.

Topoisomerase II (TOP2) poisons, such as etoposide, are potent antineoplastic drugs that also cause significant secondary toxicity to postmitotic cells. Proliferating mammalian cells express two TOP2 isoforms, TOP2A and TOP2B, whereas postmitotic cells only express TOP2B. Selectively targeting TOP2A, but not TOP2B, could thus prevent secondary toxicity in postmitotic cells. Here we report that the heat shock transcription factor, HSF1, facilitates the catalytic engagement of TOP2B on chromatin. Purified HSF1 stimulates the DNA cleavage and relaxation activity of TOP2B. Atomic force microscopy revealed that HSF1 enhances the binding of TOP2B across a range of DNA conformations. Intriguingly, HSF1 preferentially stimulates TOP2B over TOP2A, and knockdown or inhibition of HSF1 reduces the levels of catalytically engaged TOP2B but not TOP2A. Moreover, HSF1 inhibitors suppress the cytotoxicity of TOP2 poisons toward postmitotic cells without compromising their ability to kill cancer cells, revealing a strategy for minimizing the side-effects of TOP2 poison-based chemotherapy.

Cancer sonodynamic therapy (SDT) has attracted increasing attention in recent years due to its superior spatiotemporal control over chemotherapy and deeper penetration than photodynamic therapy (PDT). However, ultrasound (US) is difficult to be precisely focused on tumours. Hence, sonosensitizers that diffuse into healthy tissues may be accidently excited to generate reactive oxygen species (ROS), resulting in off-target toxicity. More importantly, current sonosensitizers exhibit high toxicity and are not biodegraded after SDT, potentially leading to systemic toxicity. Here, we developed a chitosan-oxalate hydrogel (Chi-OA) that is only stable in tumour for tumour-confined SDT. Without the need to load any traditional sonosensitizers, the hydrogel relied on the intrinsically present oxalate ions to generate ROS under US irradiation. Following intratumoural injection, the hydrogel was stable and exhibited high ROS generation yield in tumour microenvironment (TME) under US irradiation. In contrast, it degraded in healthy tissues and exhibited a lower ROS yield. This enabled SDT to be confined within the tumour and allowed the hydrogel to degrade and be cleared after SDT.

Tumor progression depends on coordinated adaptation of cancer cells and the tumor microenvironment to immune pressure, metabolic limitations, genomic instability, and biomechanical stress. While these adaptive responses have often been investigated through distinct experimental and conceptual frameworks, increasing evidence indicates that they converge on shared regulatory pathways. Calcium signaling is a fundamental regulator of cellular adaptation; however, its role in integrating tumor stress responses remains unclear. This review synthesizes emerging evidence identifying calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) as a key regulator linking calcium signaling to coordinated tumor and microenvironmental adaptation across mechanical, metabolic, replication, and immune stress responses. We discuss how CaMKK2 supports tumor ecosystem fitness through convergent tumor-intrinsic and microenvironmental programs and consider the therapeutic implications of targeting CaMKK2 and its signaling network, emphasizing context dependence, rational combination strategies, and biomarker-guided clinical translation.

Genomic instability is a hallmark of cancer, driving oncogenic mutations that enhance tumor aggressiveness and drug resistance. MYC, a master transcription factor that is deregulated in nearly all human tumors, paradoxically induces replication stress and associated DNA damage while also increasing expression of DNA repair factors and mediating resistance to DNA-damaging therapies. Emerging evidence supports a nontranscriptional role for MYC in preserving genomic integrity at sites of active transcription and protecting stalled replication forks under stress. Understanding how MYC's genotoxic and genoprotective functions diverge may reveal new therapeutic strategies for MYC-driven cancers. Here, we identify a noncanonical role of MYC in DNA damage response (DDR) through its association with DNA breaks. We show that phosphorylation at serine 62 (pS62-MYC) is crucial for the efficient recruitment of MYC to damage sites, its interaction with repair factors BRCA1 and RAD51, and effective DNA repair to support cell survival under stress. Proteomic profiling of the MYC interactome confirms a conserved interaction with components of the DDR pathway. These findings establish pS62-MYC as a key regulator of genomic stability and a potential therapeutic target in cancers.

Lipid-functionalized hybrid nanoparticles (hNPs) are promising for selective cancer delivery, due to their tunable membrane interactions. Yet, whether mimicking target membrane composition enhances recognition, and which determinants govern selectivity remains unresolved. Using coarse-grained molecular dynamics, umbrella sampling simulations, and lipid-specific decomposition analyses on 40 membrane-hNP systems, we examine how individual lipid species govern hNP interactions with mammalian-like and tumor-like bilayers. Our results showed cholesterol acts as the dominant stabilizer, generating free-energy minima and driving remodeling in both bilayers. Conversely, zwitterionic lipids showed weakened interactions, limited insertion, suppressed exchange, and entropic penalties. Tumor-like membranes amplify cholesterol's role in mediating hNP-membrane recognition, facilitating deeper insertion and lipid reorganization. Strikingly, composition-matched hNPs did not preferentially bind their corresponding membrane, whereas cholesterol-enriched formulations displayed increased affinity and selectivity. Thus, lipid-composition mimicry fails as a design principle for selective recognition. These findings provide a mechanistic basis for rational lipid selection, emphasizing complementarity over membrane mimicry.

From its early genesis, cancer is integrated with the surrounding tissue. Its very existence depends on surrounding normal tissue cells engaging with cancer cells to create an alternative tissue environment. This emerging abnormal structure becomes connected with the host organism via blood, lymphatic vessels, and neural connections. Through those connections, the cancer mass communicates and perturbs the entire organism altering various aspects of the steady-state body physiology. At early, asymptomatic stages, the induced changes within distant organs that harbor the potential to facilitate the spread of cancer are termed "premetastatic niche." Many processes involved with premetastatic changes hijack processes typical in other contexts such as development, injury, or infections, but their co-occurrence creates a new alternative physiology. The cancer to body connections not only have important consequences for the efficacy of cancer therapy but also enable cancer to evolve and adapt under the very pressure of those treatments. Furthermore, as cancer-induced changes are closely related to other physiological challenges, extrinsic perturbations such as diet, injury, and other inflammatory events have a strong impact on the tumor disease. As the disease progresses, the complex intersection of inflammatory, metabolic, and regenerative changes creates an escalating cascade of events causing cancer-related syndrome, such as cachexia, that threatens the homeostasis of the entire body and can, per se, be deadly. In this article, we will review the recent advances in the understanding of cancer as a systemic malady.

Cell migration through spatially confined microenvironments occurs in many biological processes such as embryonic development, immune surveillance, and cancer metastasis. A major bottleneck during such migration is the nucleus, which acts not only as a rigid mechanical obstacle but also as a crucial mechanosensor that modulates downstream signaling pathways. However, it remains poorly understood how nuclear deformation and mechanosensation together regulate cell migration through confined spaces. Here, we propose a three-dimensional (3D) mechanochemical model of confined nuclear translocation that integrates nuclear deformation with deformation-induced calcium signaling and subsequent regulation of cytoskeletal contractility. We show that cells undergo adaptive nuclear deformation, including nuclear envelope elongation and 3D buckling, to efficiently navigate confinements of varying sizes. There exists a biphasic relation between nuclear velocity and confinement size, arising from the interplay between nuclear deformability and mechanosensitive feedback. We demonstrate that local nuclear envelope rupture can occur under large deformation, enabling nuclear translocation through extreme confinements, as observed in prior experiments. Furthermore, we elucidate the critical roles of chromatin organization in nuclear translocation. This work reveals key mechanochemical mechanisms driving confined cell migration and provides a theoretical framework for studying nuclear dynamics across physiological and pathological contexts.

A false-negative diagnosis of cancer can lead to a delay in effective treatment and a poorer prognosis. Here, we use the example of cutaneous melanoma to examine how many years of life are lost after a false-negative diagnosis compared to a primarily correct diagnosis. From 1996 to 2015, 9,063 patients are prospectively registered in the German Central Malignant Melanoma Registry and followed up until December 2023. A false-negative diagnosis is found in 206 (2.3%) patients. The median time to correct diagnosis is 24.0 months. The 10-year recurrence-free survival probabilities are 32.9% for false-negative diagnoses and 76.2% for correct diagnoses (p < 0.001). The 10-year melanoma-specific survival probabilities are 62.1% versus 85.0% (p < 0.001). On average, each person with an initial false-negative diagnosis loses 8.2 years of life compared to people with a correct diagnosis. This high number of years of life lost raises the question of whether similar results also apply to other types of cancer.

Intravenous administration of anti-CTLA4 with anti-PD1 provides durable tumour responses but causes severe treatment-related adverse events in patients with cancer1. Intratumoural administration at lower doses but high local concentrations could enhance antitumour efficacy while minimizing systemic exposure and toxicity. Here we report the randomized multicentre phase 1b NIVIPIT trial (ClinicalTrials.gov: NCT02857569 ), which enrolled 61 patients with untreated metastatic melanoma, randomly assigned 2:1 to receive intravenous nivolumab (anti-PD1; 1 mg kg-1) combined with either intratumoural ipilimumab (anti-CTLA4; 0.3 mg kg-1) or intravenous ipilimumab (3 mg kg-1). The primary end-point was met with significantly lower incidence of grade 3 or 4 treatment-related adverse events at 6 months in the intratumoural versus intravenous arm (22.6% versus 57.1%), equivalent to anti-PD1 monotherapy. RECIST (response evaluation criteria in solid tumours) best objective response rate reached 65.7% for anti-CTLA4 injected lesions and 50% for uninjected lesions, confirming the relationship between intratumoural exposure to anti-CTLA4 and efficacy. Baseline tumour immune profiling revealed that protumoural activated regulatory T (Treg) cells and M2 macrophages predict durable clinical benefit, regardless of the anti-CTLA4 administration route. A decrease in activated intratumoural Treg cells occurred only in patients who showed durable clinical benefit, who also presented high intratumoural Fcγ receptor (FcγR) expression. Our results provide a rationale for intratumoural anti-CTLA4 strategies in oligometastatic and early-stage cancers and indicate that high intratumoural activated Treg cell and FcγR+ M2 macrophage numbers are prerequisites for efficacy of combined anti-CTLA4 and anti-PD1.

Tumor masses often exhibit heterogeneity, including escape variant clones that lack antigen-presenting machinery and/or tumor antigens, which poses a major challenge to immunotherapy. Ferroptosis, a form of regulated cell death driven by iron-dependent lipid peroxidation, has been shown to effectively induce cell death in various tumor cells. Recent studies have reported that IFN-γ suppresses the expression of System Xc-, thereby enhancing the induction of ferroptosis. Based on this, we hypothesized that combining immunotherapy with ferroptosis inducers could enhance antitumor effects against both antigen-positive and antigen-negative tumor cells. We found that combining RSL3, a ferroptosis inducer, with MART-1-specific TCR-T cells eradicates a heterogeneous tumor model consisting of human melanoma cells and their β2 microglobulin knockout counterparts. In NOG mice, this combination therapy demonstrates a significant antitumor effect against tumors with heterogeneity. These findings suggest that integrating ferroptosis inducers with immunotherapy could overcome the limitations imposed by escape variant tumor clones, offering a promising strategy for cancer treatment.

Enhancer of zeste homolog 2 (EZH2) is a methyltransferase that tri-methylates histone H3K27 as the catalytic subunit of the polycomb repressive complex 2. However, inhibition of EZH2 methyltransferase showed only variable anti-cancer efficacy, suggesting that this approach is insufficient. Here, we demonstrate a methyltransferase-independent mechanism of EZH2 wherein EZH2 interacts with inosine monophosphate dehydrogenase 2 (IMPDH2) in the cytoplasm to promote guanosine-5'-triphosphate (GTP) synthesis. Mass spectrometry identified methyltransferase-independent interactions between the EED-binding domain of EZH2 and the CBS domain of IMPDH2. EZH2 knockdown impeded IMPDH2 and reduced GTP levels, ribosome biogenesis, and cancer progression-effects reversed by guanosine. IMPDH2 knockout antagonized EZH2's tumor-promoting effects in vivo, and increased cytosolic EZH2 and IMPDH2 expression was observed in human melanomas and associated with nucleolar enlargement. EZH2-IMPDH2 complexes were also observed across multiple cancers, wherein Sappanone A (SA), which inhibits EZH2-IMPDH2 interactions, was anti-tumorigenic. These findings reveal a methyltransferase-independent oncogenic mechanism of EZH2.

Lymphocyte activation gene 3 (LAG-3) is recognized as an important inhibitory receptor on T cells, and LAG-3-targeted therapy has been approved for advanced melanoma treatment. Recently, LAG-3 has been reported to be expressed on tumor cells. However, the potential effects and underlying mechanisms of tumor cell-derived LAG-3 in mediating tumor growth and immunosuppression remain unclear. This study aimed to evaluate the role of tumor cell-derived LAG-3 within the tumor microenvironment (TME) and the efficacy of anti-LAG-3 combination therapy against melanoma. The Cancer Genome Atlas, Cancer Cell Line Encyclopedia, single-nuclear RNA sequencing database and tissue microarray analyses were conducted to detect the LAG-3 gene and protein expressions. The biological functions and underlying mechanism of tumor cell-derived LAG-3 in tumor growth and immunosuppression were investigated using phospho-array, RNA-seq, co-immunoprecipitation, flow cytometry, co-culture systems, and various tumor-bearing mouse models. Our findings revealed that LAG-3 was present in a subpopulation of cancer cells and clinical melanoma specimens associated with poorer prognosis. High LAG-3 expression on tumor cells not only contributed to the proliferation and metastasis of melanoma cells, but also promoted the recruitment of myeloid-derived suppressor cells (MDSCs) and dysfunctional CD8+ T cells into the TME. Notably, depletion of MDSCs counteracted the pro-tumor activity of tumor cell-derived LAG-3. Mechanistically, LAG-3 overexpression triggered CXCL1 and CXCL8 production of melanoma cells by enhancing TRIM28-mediated IκBα ubiquitination and degradation, which in turn activated the NF-κB pathway and fostered tumor growth and an immunosuppressive microenvironment. Intriguingly, mice bearing LAG-3-overexpressing melanoma cells showed reduced survival but responded to anti-LAG-3 antibody therapy. Furthermore, inhibiting the CXCL1/8-CXCR2 axis sensitized LAG-3-overexpressing melanoma to anti-LAG-3 antibody. This study firstly identified LAG-3 as a pro-tumor factor and a predictor for anti-LAG-3 immunotherapy efficacy. Combined CXCL1/8-CXCR2 inhibitor and anti-LAG-3 treatment may represent a promising therapeutic strategy for LAG-3-driven tumor.

Extracellular vesicles (EVs) are emerging as promising vehicles for cancer immunotherapy, yet the molecular determinants of their immunogenicity remain poorly defined. While PD-L1 expression on cancer-derived EVs has been shown to suppress immune responses, its role on immunotherapeutic EVs remains unexplored. In this study, we investigated how eliminating PD-L1 from antigen-loaded bone marrow-derived dendritic cell (BMDC) EVs affects the efficacy of EV-based cancer vaccines. We generated and characterized ovalbumin (OVA)-loaded BMDC EVs from wild-type (WT) and PD-L1-/- C57BL/6 mice. EVs were administered intravenously into WT mice in immunization experiments and in therapeutic and prophylactic B16 OVA-secreting melanoma models. Immune responses were assessed by flow cytometry, ELISpot, and ELISA, and tumour growth was monitored. Proteomic analysis confirmed high EV purity and similar protein profiles between WT and PD-L1-/- EVs, with PD-L1 being the major difference. Functionally, PD-L1-/- EVs induced significantly stronger anti-tumour responses in vivo, particularly in the prophylactic setting. Mice treated with PD-L1-/- EVs showed increased CD8+ T cell tumour infiltration, enhanced IFNγ secretion, and higher tumour rejection rates compared to WT EVs (72.7% vs. 37.5%). Additionally, the frequency of tumour-infiltrating, antigen-specific CD8+ T cells was significantly higher in PD-L1-/- EV-treated mice. In summary, BMDC-derived EVs loaded with antigen are potent immune stimulators, and removal of immune checkpoint molecules such as PD-L1 further enhances their immunogenicity. These findings support the development of engineered EVs as improved platforms for cancer immunotherapy.

Messenger RNA (mRNA) vaccine was emerging as a promising treatment for tumor immunotherapy. Highly efficient antigen-presenting ability in tumor immunotherapy through splenic dendritic cell (DC)-targeting mRNA delivery system was very important, but it remained a great challenge. In this study, a library of gemini-like ionizable lipid (termed H-type ionizable lipid, HIL) was synthesized and used for constructing mRNA-encapsulated nanoparticles (mRNA/HNPs) for in vivo mRNA delivery. Structure-activity relationship (SAR) analysis indicated that the spleen-targeting transfection efficiencies of mRNA were strongly correlated with the apparent pK a values of mRNA/HNPs after intravenous injection. After formulation screening, the optimized mRNA/HNPs based on H18 lipid (mRNA/H18NPs) with an average particle size of 124.4 ± 2.4 nm and a multilamellar concentric nanostructure were successfully prepared. Interestingly, without any ligand modification, the mOVA/H18NPs exhibited splenic DC-targeting mRNA transfection, and markedly increased the amounts of IFN-γ+ CD8+ T cells and effector memory CD8+ T cells. Furthermore, in vivo results demonstrated that mRNA/H18NPs encapsulating antigen-encoding mRNA including ovalbumin (OVA) or tyrosinase-related protein 2 (Trp2) effectively activated antigen-specific CD8+ T cells and resulted in significant antitumor efficacy in both B16-OVA or B16F10 tumor-bearing mouse models following intravenous administration. Especially, different from the mOVA/MC3-LNPs group, the mOVA/H18NPs exhibited complete inhibition of tumor progression when it used as preventative cancer vaccines in B16-OVA tumor-bearing mouse model. These findings highlighted that mRNA/H18NPs offer a promising splenic DC-targeting delivery system for mRNA vaccines.

Tumor immune evasion is greatly impacted by immune checkpoint molecules that impede the activation of costimulatory signals. Inadequate costimulatory molecules restrict the immune system's capacity to generate strong responses to malignancies. This study presents a development of a dual-nanoplex cluster (NC) formulation that effectively overcomes these issues. The formulation comprises a nanoplex (NP) encapsulating CpG ODN 1826 (CpG NP), which stimulates TLR-9 and elevates the levels of CD80/86 costimulatory molecules on antigen-presenting cells, and a NP incorporating CTLA-4 siRNA (siCTLA-4 NP), which suppresses the production of the immunological checkpoint molecule CTLA-4 on T cells. The two NPs undergo a cycloaddition reaction involving azide groups (AZ) on the NPs and alkyne groups at both termini of an MMP-2-cleavable peptide (APA) to produce a NC. The NC formation process is modulated to fabricate a flexible and deformable architecture with optimized size that maximizes passive melanoma accumulation while maintaining systemic stability, exhibiting intratumoral cleavability and substantial tumor suppression efficacy. Ex-vivo study of cell composition in spleens and tumors, along with tumor-specific antigen stimulation assays on splenocytes, suggests that NC treatment can switch the immunosuppressive tumor microenvironment into an immunocompetent state. These findings demonstrate that our NC formulation, which integrates CTLA-4 suppression with DC stimulation, potentiates immune responses against tumor tissues, providing novel insights and potential applications in the field of cancer immunotherapy.

The metabolic differences between tumor cells and normal tissue cells offer potential targets for therapeutic intervention. For instance, compared to normal tissues, tumor cells exhibit a significantly higher dependency on methionine. However, current strategies for methionine restriction have limited clinical applicability due to systemic toxicity, poor patient compliance, and insufficient tumor targeting. In this study, we developed an engineered probiotic, Met-EcN, which is able to achieve targeted consumption of methionine within the tumor microenvironment. In the B16-F10 melanoma mouse model, the combination of Met-EcN and anti-PD-L1 antibody led to a 63 % increase in the tumor inhibition rate compared to anti-PD-L1 treatment alone, as well as a 50 % improvement in mouse survival rates. Additionally, this combination significantly enhanced T cell infiltration and activation. In the MC-38 colon cancer model, treatment with Met-EcN alongside anti-PD-L1 antibody inhibited tumor growth by 84.6 %, leading to complete regression of tumors in 80 % of the mice. The depletion of methionine levels by Met-EcN leads to a reduction in methylation levels within tumor cells, which facilitates the dissociation of cGAS protein from chromatin and activates the STING signaling pathway, thereby triggering an innate immune response. This study provides a novel therapeutic approach for overcoming tumor immune resistance.

Regnase-1 is a multifunctional regulatory protein involved in immune modulation and inflammatory processes. As an endoribonuclease, it degrades mRNAs encoding pro-inflammatory cytokines, including IL-1β and IL-6. This makes it an essential factor in controlling immune cell function. Its role extends to various cell types, including keratinocytes and melanocytes, and it plays a pivotal part in maintaining skin homeostasis. Dysregulation of Regnase-1 has been implicated in various dermatological disorders, making it a significant target for understanding immune-related skin diseases. This review provides a comprehensive analysis of Regnase-1's structure, function, and its diverse roles in skin biology. Specifically, it explores how Regnase-1 influences the pathogenesis of inflammatory skin diseases such as atopic dermatitis, psoriasis, and hidradenitis suppurativa, as well as its involvement in skin cancer development. By detailing Regnase-1's regulatory mechanisms, the review aims to highlight its potential as a therapeutic target and emphasize the need for further research on its specific functions across different skin diseases and immune responses. Regnase-1 plays a crucial role in maintaining skin immune homeostasis by controlling the degradation of pro-inflammatory cytokine mRNAs, such as IL-1β, IL-6, and IL-17. It regulates inflammatory pathways in keratinocytes, melanocytes, and immune cells, preventing excessive immune activation. In atopic dermatitis, Regnase-1 deficiency leads to heightened Th2 responses and increased chemokine levels, while in psoriasis, it modulates the IL-23/Th17 axis and STAT3 pathways, impacting keratinocyte proliferation and chronic inflammation. Furthermore, in cancer, Regnase-1 acts as a tumor suppressor in melanoma by inhibiting the Akt/mTOR pathway, but its deficiency can promote squamous cell carcinoma through increased COX-2 expression. These context-specific functions make Regnase-1 a promising target for therapies that could modulate multiple inflammatory pathways, either simultaneously or specifically, offering novel approaches to treat chronic inflammatory skin disorders and cancers.

Metabolic competition and nutrient restriction in the tumor microenvironment (TME) shape the immune infiltrate in tumors and subsequently tumor immunity. In this study we used the transgenic melanoma mouse model tg(Grm1)EPv, which spontaneously develops melanoma due to the ectopic expression of the metabotropic glutamate receptor 1 (Grm1) in melanocytes to investigate if aberrant glutamate metabolism drives tumor formation and affects immune cell function. We performed liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomic analyses and RNA sequencing on tumor-free and tumor-bearing tg(Grm1)EPv tissues to characterize metabolic alterations associated with tumor progression. Flow cytometry was used to examine changes in immune cell subsets within the TME. To assess the functional relevance of glutamate metabolism, we inhibited glutathione metabolism using L-buthionine-(S,R)-sulfoximine (BSO), an inhibitor of glutamate-cysteine ligase that depletes cellular glutathione levels. LC-MS/MS-based metabolomic analyses and RNA sequencing revealed changes in glutamate and glutamine metabolism, a glycolytic shift (Warburg effect), and reduced ATP levels in advanced tumors compared with tumor-free tissue, suggesting respiratory chain dysfunction. These metabolic changes in the TME are advantageous for the tumor cells and unfavorable for immune cells, such as dendritic cells (DC). Indeed, flow cytometry analysis of myeloid subsets during tumor progression showed a decline in tumor-infiltrating conventional type 2 DC and macrophages, alongside an increase in neutrophil and monocyte populations in advanced lesions. Interference with glutamate metabolism using BSO induced immunogenic cell death, namely ferroptosis, in an tg(Grm1)EPv-derived cell line in vitro. Therefore, we evaluated the combination of this inhibitor with immunotherapy as a promising new approach for the treatment of tumors in the tg(Grm1)EPv mouse model. We observed that tumor growth could be delayed in vivo when BSO was combined with a therapy regimen boosting DC numbers and activation. This inhibition of tumor growth was supported by the infiltration of activated T cells. Overall, our findings provide novel insights into the importance of combining metabolic intervention with immunotherapy for the treatment of patients with melanoma, particularly those bearing glutamate pathway-active or immunologically cold tumors. This knowledge can drive the design of novel therapeutic strategies for patients with cancer.

Microneedle (MN) patches have emerged as a highly efficient platform for localized drug delivery, showing great promise in cancer therapy due to their ability to enable precise drug administration. However, conventional MN systems are limited by the low drug-loading capacity of their tips and primarily rely on biologically inert, nontherapeutic matrices for structural support, which restricts further gains in antitumor efficacy. Herein, we present a strategy turning toxicity into therapy by constructing palladium nanoparticle-loaded poly(vinyl alcohol)/polyethylenimine (PVA/PEI@Pd) hydrogel microneedles (PPPd-MNs), which exploit the intrinsic cytotoxicity of PEI for synergistic melanoma therapy. The PPPd-MNs efficiently catalyze the deprotection of a doxorubicin prodrug (P-DOX), enabling in situ generation of active doxorubicin (DOX). Notably, the PEI matrix serves a dual function: acting as a robust ligand to stabilize Pd catalysts and functioning as a therapeutic agent that disrupts cancer cell membranes. Both in vitro and in vivo experiments demonstrate that the combination of Pd-mediated bioorthogonal activation of DOX and PEI-induced membrane damage achieves a remarkable synergistic therapeutic outcome in a murine melanoma model, resulting in a tumor inhibition rate of up to 98%. This work repurposes the inherent cytotoxicity of the carrier material as an active therapeutic component, offering a novel paradigm for the design of high-performance bioorthogonal catalytic systems.

Immunodeficiency-associated primary central nervous system lymphoma (ID-PCNSL) represents a clinicopathologically distinct PCNSL subtype, for which large studies and prognostic models are lacking. To address this gap, the International PCNSL Collaborative Group conducted a retrospective multicenter study, integrating clinical, radiological, and pathological data from 308 ID-PCNSL cases, diagnosed at 23 participating sites in 7 countries. Preexisting immunodeficiency included administration of immunosuppressants for transplantation (41.2%) or autoimmunity (36.7%) and HIV infection (21.7%). All tumors were diffuse large B-cell lymphomas, with Epstein-Barr virus (EBV) detected in 79.2%. Immune reconstitution together with rituximab and methotrexate-based chemotherapy was associated with the highest response rates and prolonged progression-free survival, irrespective of immunodeficiency subtype and EBV status. Survival outcomes were highly variable, with a 54-month median overall survival. Multivariable Cox regression identified age (per year increment; hazard ratio [HR], 1.05 (95% confidence interval [CI], 1.02-1.07); P< .001), Karnofsky performance status (KPS) <70 (HR, 3.10; 95% CI, 1.67-5.87; P< .001), and EBV positivity (HR, 3.26; 95% CI, 1.47-7.33; P = .004) as prognostic factors for overall survival. A prognostic score was developed based on the sum of these adverse variables (age >60 years, KPS <70, EBV positivity). Stratification by this score yielded median survival times of 135, 29, and 3 months in patients with up to 1, 2, and 3 unfavorable markers (P< .0001). It allowed improved prognostic stratification of ID-PCNSL as compared with the Memorial Sloan Kettering Cancer Center and International Extranodal Lymphoma Study Group models developed for immunocompetent PCNSL. Collectively, this large international cohort defines clinicobiological features of ID-PCNSL and introduces a prognostic system with potential to guide future management.

Meningioma is a known adverse reaction of high-dose progestogens, but evidence regarding the risk associated with progestogens used as contraception is limited. To examine whether different progestogens used as hormonal contraception are associated with increased risk of meningiomas. This nested case-control study conducted over a 25-year study period from January 1, 2000, to December 31, 2024, is a Danish nationwide population-based register study, which included 3 million females aged 15 to 59 years with residence in Denmark. Meningioma cases were dentified, and for each case, 10 controls were matched on age, birthplace, and marital status and randomly selected from the cohort if considered eligible on the day of the case's meningioma diagnosis. Data were analyzed from July 10, 2025, to May 12, 2026. Use of progestogens was identified in the registers by date of dispensing or procedural records and grouped by route of administration and active substance. Exposure time was determined by redeemed daily doses or product duration. If a female switched to a different product or became pregnant, her exposure time was changed to the new exposure. The females included were allocated to their most recent use, defined as the exposure closest to the matching date. The main outcome was incident meningioma and was identified in the Danish National Cancer Register using validated International Statistical Classification of Diseases and Related Health Problems, Tenth Revision diagnoses and International Classification of Diseases for Oncology, Third Edition codes. A total of 1473 cases and 14 717 controls with a median age of 48 years (IQR, 42-53 years) were included in the nested cohort. For combined oral contraceptives, the estimated odds ratios (ORs) for the association between use of progestogens and meningioma were 1.61 (95% CI, 1.00-2.59) with cyproterone, 1.66 (95% CI, 1.31-2.10) with desogestrel, 1.58 (95% CI, 1.05-2.37) with drospirenone, 1.44 (95% CI, 1.17-1.77) with gestodene, 1.40 (95% CI, 1.12-1.76) with levonorgestrel, 1.38 (95% CI, 0.77-2.47) with norethisterone, and 1.04 (95% CI, 0.70-1.54) with norgestimate. For oral progestogen-only contraceptives, the ORs were 1.73 (95% CI, 1.17-2.56) with desogestrel and 0.95 (95% CI, 0.57-1.57) with norethisterone. For injectable medroxyprogesterone, the OR was 4.55 (95% CI, 2.19-9.45). For intrauterine devices (IUDs) with high-dose levonorgestrel, the OR was 1.58 (95% CI, 1.28-1.94), and for IUDs with low-dose levonorgestrel, the OR was 1.14 (95% CI, 0.59-2.22). Exposure within the past year was associated with the highest risk. In this case-control study conducted using data from the entire Danish population, recent use of the contraceptive progestogens cyproterone, desogestrel, drospirenone, gestodene, levonorgestrel, injectable medroxyprogesterone, and high-dose IUD was associated with increased risk of meningioma. These findings are considered relevant information for the treated women and the prescribing physicians.

In brain tumors, the immunosuppressive microenvironment leads to tumor aggressiveness and immunotherapy resistance. To identify factors that regulate the glioma immune landscape, we took advantage of the disease trajectory of the MAPK-driven glioma, pleomorphic xanthoastrocytoma (PXA), which evolves from an immune-rich low-grade tumor to an aggressive glioma. Using a multiomics approach that included single-nucleus RNA sequencing (snRNA-seq), spatial transcriptomics, and spatial proteomics, a cohort of 38 tumors, including 10 longitudinal pairs, was profiled. Tumor progression was associated with transition to a more proliferative progenitor state and increased hypoxia. Hypoxic reprogramming of the immune landscape included increased TGF-β signaling and altered chemokine signaling, including decreased CXCL14 and CXCL16-CXCR6 signaling important in immune cell recruitment and activation. The resulting immune landscape was spatially reorganized with reduced HLA Class II expression, reduced CD8+ and CD4⁺ T cells, and dominated by immunosuppressive myeloid cells. Hypoxia and chemokine signaling emerged as key components of the glioma-immune landscape that evolve with tumor progression. These findings identify potential therapeutic opportunities in PXA, and they argue that optimal immunomodulatory strategies in glioma will differ along the disease trajectory.

CAR-T therapy is effective in hematologic cancers but faces challenges in solid tumors due to antigen heterogeneity and an immunosuppressive tumor microenvironment (TME). Systemic CTLA-4 blockade enhances immunity but often causes severe adverse events. To overcome these limitations, we developed a dual-modular nanobody-based CAR-T platform targeting fibroblast activation protein (FAP) on cancer-associated fibroblasts and locally releasing an anti-CTLA-4 nanobody within the tumor stroma. FAP/CTLA-4 dual-module CAR-T cells were generated and assessed in vitro for antigen-specific cytotoxicity, cytokine release, and exhaustion. Antitumor efficacy was evaluated in xenograft models, measuring tumor growth, survival, and T-cell infiltration (Ethics Approval Number: 202001011). One patient with refractory glioblastoma received intrathecal infusion; clinical response, cerebrospinal fluid (CSF) cytokines (Ethics Approval Number 2022-0553-01), and safety were monitored. Tumor and immune microenvironment changes were analyzed via transcriptomic sequencing and multiplex immunofluorescence staining. In vitro, engineered CAR-T cells showed potent cytotoxicity, cytokine production, and reduced exhaustion. In vivo, they induced tumor regression, prolonged survival, and increased T-cell infiltration. In the glioblastoma patient, intrathecal administration resulted in disease stabilization, elevated CSF cytokines, and a favorable safety profile. Transcriptomic sequencing and multiplex immunofluorescence staining indicated TME remodeling toward an immunologically active state. FAP/CTLA-4 DMN CAR-T overcomes the immunosuppressive solid tumor microenvironment through localized immunomodulation, demonstrating promising efficacy in preclinical models and a patient with refractory glioblastoma.

Acetylcholine was identified over a century ago with early studies focusing on its release from neuronal systems; while a growing interest in non-neuronal systems has emerged, little is known about acetylcholine release from glioblastoma cells, and its mechanism remains unexplored. Here, we quantified the real-time acetylcholine release from live, single U-87 MG glioblastoma cells and investigated its mechanistic framework through the combination of nanoelectrodes with an interface between two immiscible electrolyte solutions and nano-resolved positioning using scanning electrochemical microscopy. Complementary to conventional solid electrodes, this measuring technique enabled quantitative and selective detection of redox-inactive acetylcholine. We hypothesized that U-87 MG glioblastoma cells may release acetylcholine, considering they express cholinergic markers and exhibited a ∼3-fold increase in intracellular calcium activity following KCl stimulation. We achieved the highly sensitive local measurement of the acetylcholine release by positioning the electrode ∼200 nm away from the glioblastoma cell surface. The release of acetylcholine from U-87 MG glioblastoma cells in the substantially reduced calcium environment or with the treatment of vesamicol, an inhibitor of vesicular acetylcholine transporter, only decreased slightly from the control condition. This suggests the presence of a possible nonvesicular mechanism of acetylcholine release in cancer cells, differing from neurons. The unprecedented findings on the acetylcholine release mechanism in U-87 MG glioblastoma cells presented here provide new insights in understanding the role of acetylcholine in glioblastoma disease progression.

Diffuse intrinsic pontine glioma (DIPG) is a highly aggressive and fatal pediatric brainstem tumor, characterized by rapid progression and extremely poor prognosis. Its infiltrative growth within the pons, combined with the critical function and inaccessibility of the brainstem, makes surgical resection different and significantly limits therapeutic options. Moreover, the molecular and cellular heterogeneity of DIPG complicates therapeutic development, and despite decades of research, the current standard of care remains limited to palliative radiotherapy. In recent years, tumor immunology in DIPG has gained attention, as its immunosuppressive microenvironment, resulting in an immunologically 'cold' tumor, presents both a challenge and an opportunity for advancing immunotherapeutic strategies. In this review, we focus on outlining what is known about the developmental biology, pathological hallmarks, and immunosuppressive tumor microenvironment of DIPG, including immune infiltration and function of both immunosuppressive cells and effector NK and T cells. We also summarize recent advances in the therapeutic landscape of DIPG, including targeted therapies (epigenetic modulators, receptor tyrosine kinase inhibitors, cell cycle and DNA damage response inhibitors, metabolic immune modulators), immunotherapies (adoptive cell therapy, immune checkpoint inhibitors, vaccine therapy and oncolytic virotherapy), and novel delivery approaches currently under preclinical and clinical investigation. Emerging trends highlight the potential of combination therapies to enhance treatment efficacy. A deeper understanding of DIPG biology and tumor microenvironment is critical for advancing more effective therapies for this aggressive disease among these vulnerable populations.

In cancer cells, pyruvate generated through aerobic glycolysis is preferentially converted into lactate. Here, we examined the impact of aerobic glycolysis on innate immune regulation within the tumor microenvironment. We identified lactate as a potent suppressor of stimulator of interferon genes (STING)-mediated innate immune signaling. Lactate directly bound the cyclic GMP-AMP (cGAMP)-binding domain of STING, thereby inhibiting cGAMP binding, STING activation, and interferon regulatory factor 3 (IRF3)-dependent cytokine expression. Mechanistically, activation of epidermal growth factor receptor (EGFR) promoted phosphorylation of lactate dehydrogenase A (LDHA) by pyruvate kinase M2 (PKM2) at serine 161 (S161). This increased LDHA activity and lactate production, which suppressed immune cell infiltration and promoted tumor growth. Pharmacological PKM2 inhibition relieved lactate-mediated STING suppression, reduced tumor immune evasion, and synergized with anti-PD-1 therapy. In human glioblastoma, elevated LDHA S161 phosphorylation correlated with reduced STING activation, diminished cytotoxic immune cell infiltration, and poor survival. Thus, oncogenic signaling directs PKM2-generated pyruvate toward LDHA-mediated lactate production, which directly inhibits STING to silence innate immune activation.

Pediatric high-grade gliomas (pHGG) are the leading cause of childhood cancer-related deaths. Those arising in the midline harboring a lysine to methionine substitution at position 27 in histone 3 (H3K27M), termed diffuse midline glioma (or DIPG when occurring in the pons), are particularly deadly and in need of additional therapeutic options. Our group and others have found upregulation of the RAS/MAPK pathway across HGGs, including DIPG; however, RAS is notoriously difficult to target therapeutically, with no approved drugs that can target non-mutant RAS proteins. We leveraged a pan-RAS-cleaving biologic (RRSP-DTB) to define RAS dependency in DIPG and employed proteomic profiling of patient-derived tumors to identify enriched cell-surface receptors for tumor-selective targeting. RRSP was re-engineered to engage the lead candidate receptor, tumor endothelial marker 8 (TEM8/ANTXR1), generating a targeted RAS-degrading biologic evaluated in vitro and in orthotopic DIPG xenograft models. Pan-RAS cleavage revealed DIPG to be critically dependent on RAS/MAPK signaling compared to other RAS/MAPK HGGs. Targeting RRSP to TEM8 enabled efficient intracellular delivery, resulting in complete RAS ablation and apoptotic cell death in patient-derived DIPG cells at low picomolar concentrations. In vivo, intracerebroventricular delivery reduced leptomeningeal disease burden (p = 0.04), while convection-enhanced delivery extended survival in orthotopic DIPG models (median survival 68 vs. 57 days; p = 0.016). We demonstrate RAS-dependency in DIPG targetable by RRSP-DTT-TEM8, a novel, first-in-class pan-RAS biotherapeutic, supporting its potential as a therapeutic strategy for this otherwise untreatable disease.

TEAD transcription factors enable the oncogenic activity of deregulated Hippo signaling and are a promising therapeutic target in oncology. Targeting the TEAD lipid pocket is an established path to inhibit the oncogenic activities of cofactors YAP and TAZ. Here we present two pan-TEAD inhibitors, GNE-8025 and its in vivo brain-penetrant derivative GNE-2181, that covalently bind the lipid pocket at a conserved cysteine. Both small molecules show growth inhibition of YAP-driven tumor cells in vitro and in vivo. Moreover, we show that GNE-8025 increases the activity of a broad range of MAPK pathway inhibitors in vitro as well as the KRASG12C inhibitor Divarasib both in vitro and in vivo. In addition, GNE-2181 inhibits growth of an intracranial tumor model in vivo. Altogether we present a next-generation class of TEAD inhibitors representing a significant advancement towards potent, specific, and effective Hippo-targeting cancer therapies.

Precision oncology aims to tailor treatment according to tumor-specific molecular alterations, but the success of aberration-guided therapies has been limited in clinical trials. Here, we develop an integrated whole-genome and transcriptome workflow to systematically distinguish functionally credible, predictive driver aberrations from non-functional alterations across all classes of genomic events. We applied the integrated omics workflow to 335 patients with ovarian high-grade serous carcinoma (HGSC) enrolled in the observational DECIDER study. Tumor samples were collected from multiple cancer sites as part of the standard cancer care. DNA and RNA were extracted together from snap-frozen tumor samples and sent to whole-genome and transcriptome sequencing. Sequencing data were processed with the Anduril 2 pipeline for detection and validation of short somatic changes and with the HMW toolkit and the nf-core/rnafusion pipeline for assessment of structural changes. Aberration-specific drug sensitivity was tested in patient-derived organoids with a drug screen combining targeted agents and chemotherapy. Using an agnostic integrated omics analysis, we identified clinically relevant ESCAT Tier II-III alterations in more than 40% of the patients, even though 58% of all nominally pathogenic variants proved to be false positives. Credible aberrations were predominantly clonal, detected across anatomical sites, and preserved from diagnosis to relapse, indicating early establishment during tumor evolution. The most recurrent actionable event was NF1 deficiency, which was associated with a robust transcriptional footprint and marked sensitivity to KRAS- and MEK-inhibition in patient-derived organoids. Notably, integrated DNA-RNA analysis enabled discrimination of treatment-guiding aberrations from false-positive findings that would otherwise misinform treatment selection and confound clinical trial outcomes. Our findings provide a strategy for more reliable biomarker detection in precision oncology, inform biomarker-guided clinical trial design, and reveal unexploited therapeutic vulnerabilities in HGSC.

The clinical efficacy and immune modulation of neoadjuvant immunochemotherapy for epithelial ovarian cancer (EOC) remain uncertain. To clarify these effects, the NAIVE trial (NCT04815408), a prospective phase II study, evaluated the efficacy of neoadjuvant platinum-based chemotherapy with tislelizumab (NACI) in comparison with chemotherapy alone (NAC) in patients with FIGO IIIC-IV EOC. The primary endpoint was the 1-year progression-free survival (PFS) rate; the secondary endpoints included PFS, R0 resection, clinical and pathological responses, and safety. Between April 2021 and July 2024, 25 patients were included in the final analysis. After a median follow-up period of 30.7 months, NACI was associated with numerically prolonged progression-free survival (27.2 vs. 21.8 months; HR = 0.44, 95% CI 0.15-1.26; P = 0.127), with higher 1-year (92.3% vs. 83.3%) and 2-year (62.3% vs. 31.8%) PFS rates than NAC. NACI also yielded superior tumor responses, including higher ORR (69.2% vs. 58.3%), more R0 resections, and increased CRS3 rates. No unexpected safety signals emerged, and immune-related adverse events were manageable. Immune profiling techniques, such as single-cell RNA sequencing and CyTOF, identified distinct signatures. NACI responders demonstrated heightened CD8+ T-cell-mediated toxicity and metabolic fitness, whereas nonresponders exhibited exhaustion phenotypes. Furthermore, the response correlated with enrichment of the CXCL13+Th1-GC B-cell-tertiary lymphoid structure (TLS) axis, accompanied by increased TLS activity and differentiation to IgG-producing plasma cells. Overall, NACI yielded PFS trends with acceptable safety, but without statistical significance. The mechanistic analyses highlight the role of the CXCL13+Th1-GC B-cell-TLS axis as a potential biomarker and driver of therapeutic response.

Elevated neutrophil-to-lymphocyte ratio (NLR) has been associated with platinum resistance and poor outcomes in high-grade serous ovarian cancer (HGSOC), but the biological basis of this association remains unclear. We retrospectively analyzed 434 patients with HGSOC treated with platinum-based chemotherapy and found that elevated NLR was an independent predictor of platinum resistance. To further investigate the cellular basis of this association, we performed immunohistochemistry, proteomic profiling, and single-cell RNA sequencing on patient samples. These analyses identified a distinct subpopulation of V-domain immunoglobulin suppressor of T cell activation-positive neutrophils (VISTA⁺Neus) enriched in resistant tumors. High VISTA⁺Neus density was linked to shorter progression-free survival and showed greater predictive value for resistance than total neutrophils. Spatial and multicolor immunohistochemistry analyses further showed that VISTA⁺Neus were associated with reduced CD8⁺T-cell infiltration and cytotoxic features. Functional validation in an immunocompetent mouse model showed that anti-VISTA plus cisplatin reduced tumor growth, decreased neutrophil abundance, and increased CD8⁺T-cell infiltration and granzyme B expression. CD8⁺ T-cell depletion markedly attenuated the therapeutic benefit of the combination. These findings support VISTA⁺Neus as a potential immunosuppressive neutrophil subset associated with platinum resistance and CD8⁺T-cell suppression in HGSOC, and support further investigation of VISTA-targeted strategies and the potential biomarker value of VISTA⁺Neus.

Patients with advanced ovarian cancer who fasted before and after chemotherapy treatments had lower insulin levels and better outcomes compared with those who ate normally, according to results from a recent study.

Low-grade serous carcinoma of the ovary (LGSOC) is relatively resistant to chemotherapy. Given its biological parallels to hormone receptor-positive breast cancer, including responsiveness to anti-estrogen therapies, we conducted a pilot phase II study to assess clinical benefit of neoadjuvant fulvestrant and abemaciclib in women with advanced, unresectable LGSOC (NCT03531645). Imaging assessments were performed every 8 weeks until resectable. The primary endpoint was clinical benefit rate (CBR). Exploratory objectives included evaluation of safety profile, accessing biomarkers related to clinical benefit, and description of tumor phenotypic changes. Fifteen patients were enrolled and evaluable for efficacy. CBR was 100%, with 1/15 patients (7%) achieving complete response (CR), 8/15 patients (53%) achieving partial response (PR), and 6/15 (40%) exhibiting stable disease (SD). Interval cytoreductive surgery (ICS) was performed in 9 patients (60%), with 7 (78%) achieving complete or optimal resection. Fourteen tumor samples underwent transcriptomic and proteomic profiling. Tumors from long-term survivors showed significantly higher baseline expression of cell-cycle-related programs and estrogen signaling-related genes, both suppressed upon treatment. Neoadjuvant fulvestrant and abemaciclib was well tolerated, achieved high response rates, and enabled optimal surgical resection in most patients. Tumor proliferative activity and estrogen signaling dependency may predict therapy response.

Residual disease after cytoreductive surgery is a dominant, surgeon-modifiable determinant of outcome in advanced ovarian cancer, yet white-light inspection cannot reliably identify microscopic implants, plaque-like deposits, or therapy-altered fibrotic foci across complex peritoneal surfaces. Intraoperative molecular imaging aims to close this visibility gap by translating tumor-associated biology into real-time signal that can prompt additional resection, direct sampling, and support intraoperative decision-making. Across probe classes, folate receptor-α-targeted agents represent the most clinically mature approach, while activatable tracers and multimodal platforms are expanding capabilities beyond superficial visualization. However, improved detection alone does not establish patient benefit. We therefore frame molecular guidance as a workflow intervention and organize evidence along a clinically oriented hierarchy-lesion detection, decision impact, and patient outcomes-highlighting the need for standardized acquisition/quantification and pathology-linked validation. We propose pragmatic, trial-compatible definitions for pathology-confirmed completeness and prespecified molecular field clearance, and discuss how AI-assisted interpretation, multimodality, and radionuclide-enabled strategies could strengthen reproducibility and extend management toward microscopic residual disease.

The Hippo pathway is a key regulator of development, regeneration, tissue homeostasis, and organ size, and its dysregulation promotes tumorigenesis. However, the precise mechanisms of its regulation in both normal physiology and cancer remain incompletely understood. Here, we identify STK38 and STK38L (also known as NDR1 and NDR2), previously proposed as redundant kinases of LATS, as negative regulators of the Hippo pathway. STK38/L inhibit LATS by competitively binding to MOB1 and disrupting the LATS-MOB1 complex, a process independent of their kinase activity. This inhibitory mechanism is evolutionarily conserved, as the Drosophila ortholog Tricornered similarly impairs Warts-Mats complex formation, resulting in enlarged fly wing size. Pathologically, STK38L is highly expressed in ovarian cancer and required for ovarian tumor growth, and its amplification correlates with YAP activation and increased tumor sensitivity to TEAD inhibitors. Taken together, our study reveals a conserved role of STK38/L in Hippo pathway regulation, providing new insights into Hippo-dependent growth control and cancer development.

While immune checkpoint inhibitors (ICIs) have failed to improve outcomes in unselected ovarian cancer populations, objective responses are observed in a minority of ovarian clear cell carcinoma (OCCC) cases, implying biological heterogeneity and a yet-undefined immunologically responsive subset within this histotype. We performed immunohistochemical profiling of tumor-infiltrating immune cells and analyzed transcriptomic data from human OCCC cohorts. Functional studies were conducted using an immunocompetent syngeneic OCCC mouse model to assess the effects of IL-17 on tumor cell inflammatory signaling, immune microenvironment remodeling, and responsiveness to immune checkpoint blockade, including single-cell RNA sequencing of tumor-infiltrating T cells. OCCC exhibited an immune-sparse tumor microenvironment with relative enrichment of CD4⁺ T cells. RORC expression was elevated in OCCC but showed intertumoral heterogeneity. In the transcriptome data (n = 180), an IL17Ahigh subset (5%), enriched within the RORChigh fraction, exhibited a T cell-inflamed gene expression profile independent of microsatellite instability and tumor mutational burden, yet was not associated with survival. Mechanistically, IL-17 directly activated NF-κB-dependent inflammatory programs in OCCC tumor cells, inducing cytokines and chemokines involved in T-cell recruitment and activation. In the syngeneic model, IL-17 exposure increased intratumoral CD4⁺ and CD8⁺ T-cell infiltration and activation. Single-cell profiling further revealed expansion of Th17/Tfh-like CD4⁺ T cells and cytotoxic, non-terminally exhausted CD8⁺ T cells. Consistent with these changes, anti-PD-L1 therapy improved survival in Th17-biased partial chimera mice. IL-17-responsive, tumor cell-intrinsic inflammatory programming remodels the tumor immune microenvironment toward an immunotherapy-permissive state. These findings establish IL-17-responsive tumor cell inflammatory programming as a mechanistic axis shaping immune checkpoint sensitivity and provide a rationale for biomarker-guided immunotherapy strategies.

This exploratory study aimed to evaluate the clinical feasibility and added diagnostic value of the heterodimeric agent 68 Ga-NOTA-3P-TATE-RGD, targeting both SSTR2 and integrin α v β 3 , in patients with radioactive-iodine-negative differentiated thyroid cancer (DTC), through a comparison with 18 F-FDG PET/CT. Twenty-two patients with radioactive-iodine-negative DTC underwent both 68 Ga-NOTA-3P-TATE-RGD and 18 F-FDG PET/CT within 1 week. Diagnostic performance and semiquantitative parameters were compared between the two agents. Based on the collected gene mutation data, a preliminary analysis was conducted on differences in PET uptake among different gene mutation types. A total of 265 and 295 lesions were detected on 68 Ga-NOTA-3P-TATE-RGD and 18 F-FDG PET/CT, respectively. 68 Ga-NOTA-3P-TATE-RGD detected 45 exclusive lesions (12.71%), predominantly in bone (6 lesions) and brain (5 lesions), while 18 F-FDG identified 75 exclusive lesions in lymph nodes (24 lesions) and lungs (51 lesions). The dual-targeted agent exhibited superior total specificity (92.3% vs 70.6%) and significantly higher tumor-background ratio for bone metastases [3.4 (2.6, 4.0) vs 2.3 (1.6, 2.9), P = 0.008]. For PET comparison of different gene mutations, 68 Ga-NOTA-3P-TATE-RGD showed highest uptake in TERT promoter-mutant lesions ( P < 0.01), while 18 F-FDG had maximal uptake in BRAF-mutant lesions ( P < 0.01). 68 Ga-NOTA-3P-TATE-RGD PET/CT is feasible for radioactive-iodine-negative DTC, providing complementary diagnostic information to 18 F-FDG with enhanced detection of bone and brain metastases. The intense uptake observed in lesions with TERT-associated aggressive subtypes supports potential utility for specific disease identification.

In differentiated thyroid cancer (DTC), the incidence of bone metastases of papillary thyroid cancer (PTC) is relatively low, with primary sites including the spine, pelvis, and ribs. While metastasis to the phalangeal bones is particularly rare. Here we reported a case of right ring figure metastasis in a 53-year-old woman with PTC. The right ring figure lesion was confirmed to be metastatic PTC by pathologic examination and immunohistochemistry.

A 55-year-old man with high-risk papillary thyroid cancer underwent a second round of radioiodine therapy due to lung and lymph node metastases. A follow-up whole-body scan showed a new focal area of radioiodine uptake in the right lower limb, which was absent in previous scan and remained despite skin decontamination. Upon examination, a posttraumatic scab was found on the right leg from a fall 5 days prior. SPECT/CT imaging and scab healing during follow-up effectively ruled out bone and skin metastases. This case underscores the need to recognize false-positive radioiodine uptake in differentiated thyroid cancer for improved diagnostic accuracy and patient care.

Dysregulation of Polycomb Repressive Complex 2 (PRC2) contributes to cancer. Of its catalytic subunits, Enhancer of Zeste (EZH) 1 and EZH2, EZH2 mutations are extensively studied, but the role of EZH1 in cancer remains largely unexplored. Here, we investigate the thyroid cancer-associated EZH1Q571R mutation and uncover a mechanism that extends beyond catalytic gain of function. Using biochemical, single-molecule, epigenomic, and transcriptomic analyses, we demonstrated that EZH1Q571R significantly enhances chromatin compaction and stimulates PRC2-EZH1 catalytic activity, thereby rewiring PRC2-chromatin interactions. This altered engagement enables PRC2 activity within H3K36me2-marked chromatin typically refractory to H3K27 methylation, leading to widespread epigenetic and transcriptional reprogramming. Notably, enhanced chromatin compaction is observed with EZH1Q571R but not with the corresponding EZH2Q570R mutation, indicating an EZH1-specific mechanism. Functionally, EZH1Q571R accelerated tumor growth in vivo and induced pronounced micronuclei formation ex vivo, reflecting aberrant chromatin compaction and genome instability. Together, our study demonstrates that EZH1Q571R promotes a permissive epigenomic landscape for the progression of follicular thyroid cancer.

Oncocytic (Hürthle cell) carcinoma of the thyroid (OCT) is characterized by widespread loss of heterozygosity (LOH), mitochondrial accumulation, and recurrent mitochondrial DNA mutations leading to impairment of complex I. Here, we establish and characterize a novel OCT cell line, UT946, which displays severe mitochondrial electron transport chain dysfunction and a Warburg metabolic phenotype. Using a series of cytoplasmic hybrids, we establish that the complex I defect in UT946 stems from a nuclear-encoded loss-of-function mutation in the complex I subunit NDUFS1. To our surprise, the mutation in NDUFS1 was inherited as a recessive germline allele that underwent LOH in the tumor to expose functional loss of complex I. A reanalysis of 91 OCT tumor genomes revealed that LOH-driven exposure of recessive germline mutations in complex I subunits was a recurrent mechanism underlying complex I inactivation in OCT. These findings unveil a previously unidentified germline-driven mechanism of complex I loss and metabolic reprogramming in cancer and provide further evidence of the selective pressure for complex I impairment in OCT.

Lymph node (LN) metastases are a major driver of mortality across solid cancers, including thyroid carcinomas, which are known for high rates of nodal colonization. To elucidate the determinants of nodal spread, we isolated tumor-infiltrating leukocytes from primary thyroid tumors and matched metastatic LNs for single-cell RNA sequencing with validation by multiplex immunohistochemistry. Comparing the microenvironmental alterations between primary tumors and their LNs, we found that thyrocytes and tumor-associated macrophages down-regulate the expression of multiple inflammatory cytokine receptors, including TNFRSF12A and CX3CR1, upon LN colonization. LNs were associated with the induction of regulatory T cells to suppress T cell-mediated cytotoxicity compared to matched primary tumors. Notably, tumor-infiltrating lymphocytes within LNs demonstrated increased expression of activation markers, including interleukin-7 receptor (IL7R). High LN expression of IL7R was significantly correlated with improved outcomes and can serve as a biomarker in this heterogeneous disease. Our findings on the dynamic equilibrium within LN metastases may offer conserved mechanisms for nodal colonization across solid tumors.

Engineered T-cell therapies, such as chimeric antigen receptor (CAR)- and T-cell receptor (TCR)-based approaches, have transformed outcomes in hematologic malignancies, yet their efficacy in solid tumors remains limited by tumor antigen escape, immunosuppressive microenvironments, and insufficient activation of CAR or TCR signaling. To overcome these barriers, we developed an intercellular adhesion molecule 1 (ICAM1)-specific chimeric costimulatory receptor (ICCR) engineered for expression in T cells to augment their activation. ICAM1 is broadly expressed across solid tumors and is further upregulated by IFNγ released during early T-cell engagement, creating a feed-forward loop that reinforces tumor recognition. ICCR engagement with ICAM1 triggered NF-κB signaling independently of TCR-peptide (p)/MHC engagement; however, full T-cell activation and cytotoxic function remained dependent on intact TCR signaling. In primary T cells, ICCR increased proliferation, cytokine production, and cytotoxicity, resulting in improved tumor control in two anaplastic thyroid cancer xenograft models treated with allogeneic or autologous ICCR-T cells. Mechanistically, ICCR strengthened tumor cell engagement, promoted selection and expansion of tumor-specific TCR clonotypes, and amplified downstream signaling pathways. These findings identify ICCR as a strategy that leverages an immune synapse-mimetic mechanism to enhance the function of low-activity tumor-specific TCRs and improve T-cell responses in solid tumor microenvironments.

We screened 424 natural compounds, including many marketed as dietary supplements, to assess their antioxidant and thyroidal effects. In rat PCCL3 thyroid follicular cells, we performed parallel primary screens for antioxidant response element (ARE) activation, thyroglobulin (Tg) promoter activity, sodium/iodide symporter (NIS) promoter activity and iodide uptake. Hits were tested in downstream analyses, including ARE activation in Nrf2-knockout cells, an Nrf1 stabilization reporter, endogenous Nqo1, Tg and NIS mRNA and protein levels, and relationships between these readouts. Of 424 compounds, 69 induced ARE activity in wild-type cells; most also activated the ARE in Nrf2-knockout cells, and only 8 showed a profile consistent with predominantly Nrf2-dependent ARE activation. Several Nrf2-independent ARE inducers also stabilized an Nrf1-based reporter. Thirty compounds activated the Tg promoter and 26 significantly modulated iodide uptake, with only partial concordance between promoter activity, mRNA/protein expression and iodide uptake. Notable hits included genistein and apigenin, which showed balanced ARE and Tg induction together with NIS upregulation, supporting further evaluation in hypothyroidism and redifferentiation; luteolin, chrysin and diosmetin, which enhanced NIS expression and/or iodide uptake, highlighting potential as redifferentiation or adjunctive agents in thyroid cancer; emodin and gamma-oryzanol, which increased iodide uptake, suggesting utility in hypothyroidism or iodine deficiency; and bavachin, which showed the strongest iodide uptake induction without markedly increasing NIS protein, warranting mechanistic follow-up. Thyroid-relevant bioactivity was enriched among dietary supplements. Many compounds displayed complex Nrf2-dependent and -independent actions that can confer indication-specific thyroid benefits or risks, emphasizing the need for thyroid-focused evaluation of antioxidant and natural supplements.

Oral squamous cell carcinoma (OSCC) is one of the most common cancers in the head and neck. Immunotherapy has emerged as a promising treatment option for metastatic OSCC because of its potential clinical benefits; however, its effectiveness is limited by the immunosuppressive tumor microenvironment (TME), short-lived responses, and poor infiltration. To overcome these issues, we developed a strategy that combines photothermal therapy (PTT) with chimeric antigen receptor (CAR)-T cell immunotherapy. The prepared conjugated polymer nanoparticles (CPNPs) serve as efficient near-infrared-II (NIR-II) photothermal agents, enabling localized PTT and triggering strong immunogenic cell death (ICD) to activate T cells. Moreover, we engineered a lymph node-targeting nanosystem (ApoA1@PNPs) to improve in vivo production of CAR-T cells. Mucin 1 (MUC1)-specific CAR-T cells were designed to enhance tumor antigen recognition. This combined approach helps CAR-T cells reach primary tumor sites more effectively and induces long-lasting systemic immunity. By addressing the main limitations of traditional CAR-T therapy in OSCC, our integrated PTT/CAR-T strategy offers a potential therapeutic approach with significant clinical potential. This dual method aims to improve patient outcomes by achieving better tumor control.

Oral squamous cell carcinoma (OSCC) lacks effective low-toxicity treatments. Chemodynamic therapy (CDT) offers a tumor-specific approach by converting hydrogen peroxide into toxic radicals. However, its efficacy is limited by insufficient H2O2, high glutathione (GSH) levels that neutralize the radicals, and reliance on a single cell death pathway. Herein, we report a precisely programmable catalytic platform consisting of Ru single atoms anchored on a CuTi layered double hydroxide (Ru CuTi-LDH) nanozyme. The Ru sites not endow the nanozyme with superoxide dismutase (SOD)-like activity and enable precise control over its catalytic functions, which also include peroxidase (POD), catalase (CAT), and glutathione peroxidase (GPx). Together, these features orchestrate a precise cascade reaction to amplify therapeutic efficacy for OSCC. Light-triggered superoxide radicals (•O2-) are converted to H2O2 by Ru sites, fueling Fenton-like reactions at Cu centers that generate cytotoxic hydroxyl radicals (•OH). Meanwhile, Ru CuTi-LDH depletes GSH and generates O2 to alleviate tumor hypoxia. This chemical reprogramming amplifies oxidative damage and sensitizes tumor cells to cuproptosis. Additionally, endoplasmic reticulum (ER) stress triggered by the cascade activates paraptosis, establishing three distinct cell death pathways simultaneously. This approach achieved 84.7% tumor inhibition and prolonged survival in an orthotopic OSCC model. This work presents a chemical strategy that addresses fundamental CDT limitations through cascade catalysis with atomic-level tunability.

Microplastics (MPs, <5 mm) and nanoplastics (NPs, <1 μm) are pervasive pollutants increasingly recognized as emerging threats to human health. While their systemic impacts on the gastrointestinal, respiratory, reproductive, and immune systems are well documented, their relevance to oral health has received limited attention. The oral cavity represents both a primary site of exposure, via ingestion, inhalation, and contact with dental and personal care products, and a sensitive biological interface where local toxicity may arise. This review synthesizes evidence on oral sources of MPs/NPs, including toothbrushes, toothpastes, orthodontic appliances, restorative composites, prostheses, implants, and impression materials. We highlight potential links to oral diseases such as gingivitis, periodontitis, peri-implantitis, denture stomatitis, oral cancer, and xerostomia. Mechanistic studies demonstrate that MPs/NPs trigger oxidative stress, inflammatory signaling, immune dysregulation, microbiome disturbances, and endocrine disruption, with implications for both local pathology and systemic dissemination. Once crossing oral and gastrointestinal barriers, these particles can accumulate in distant organs, exacerbating chronic inflammatory and metabolic disorders. We conclude by outlining key research gaps, emphasizing the need for advanced detection methods, sustainable dental materials, and translational studies to clarify clinical relevance. Collectively, this review underscores the oral cavity as a critical but underexplored interface for plastic particle exposure.

Head and neck cancer (HNC) presents significant clinical challenges due to its complex tumor biology, heterogeneous microenvironment, and limited therapeutic outcomes. Hydrogels, an innovative biopolymeric material, have gained considerable attention in both basic and translational oncology owing to their biocompatibility, biodegradability, tunable physicochemical properties, and ability to recapitulate the native tissue environment. This review provides a comprehensive overview of HNC epidemiology, molecular and cellular mechanisms, current and emerging therapeutic approaches, and the defining features of the tumor microenvironment. We then highlight diverse hydrogel systems under development, detailing their structural characteristics and biomedical applications. Hydrogel-based platforms have advanced tumor modeling by enabling three-dimensional (3D) culture systems that more accurately reflect in vivo conditions compared to conventional two-dimensional (2D) models. Cutting-edge technologies, including 3D bioprinting and tumor-on-a-chip microfluidic systems, offer new opportunities for precision modeling and drug testing in HNC. In addition, hydrogels are being engineered for therapeutic applications, such as localized and sustained drug delivery, immunomodulation, and tissue regeneration following tumor resection. Finally, we critically examine the challenges and limitations of current hydrogel-based approaches and outline future directions for their integration into HNC research and clinical practice.

Oral squamous cell carcinoma (OSCC) remains a therapeutic challenge due to its aggressive metabolism-driven progression and immunosuppressive tumor microenvironment (TME). Through proteomic and transcriptomic analyses, we identified HNRNPC as a novel driver of OSCC progression via aberrant glycolysis and TGF-β-mediated differentiation of ITGA1+ myofibroblastic cancer-associated fibroblasts (myCAFs), which directly induce CD8+ T cell exhaustion. Given the lack of strategies that simultaneously target both tumor and stromal compartments, we engineered a theranostic solution: Near-infrared-II (NIR-II) imaging-guided aggregation-induced emission nanoparticles (AIE NPs) dually conjugated with antibodies against HNRNPC and ITGA1 (NPs-H-I). Studies using molecular experiments, single-nucleus sequencing (snRNA-seq), allograft models, and clinical specimens confirmed that HNRNPC-high OSCCs are enriched with ITGA1+ myCAFs and exhausted CD8+ T cells, correlating with poor patient survival. The developed NPs-H-I demonstrated specific accumulation in tumors, enabling precise NIR-II imaging. Upon 808 nm laser irradiation, NPs-H-I elicited potent photodynamic therapy (PDT), selectively eliminating HNRNPC-high tumor cells and ITGA1+ myCAFs, which robustly inhibited tumor growth and restored anti-tumor immunity in murine models. Our work not only delineates the HNRNPC-ITGA1 axis as a novel metabolic-immune checkpoint in OSCC but also establishes a versatile and translatable precision medicine platform capable of disrupting this pathway for effective combination therapy.

Oral cancer and oral potentially malignant disorders (OPMDs) remain a significant challenge in diagnosis and therapy, primarily due to inherent limitations in early detection, targeted treatment, and postoperative rehabilitation. Conventional diagnostic and therapeutic modalities often lack sufficient sensitivity, specificity, and effectiveness in restoring oral function. Biomaterials including nanoparticles, hydrogels, and scaffolds, offer versatile solutions by virtue of their tuneable properties, biocompatibility, and versatility in drug delivery and tissue engineering. However, their clinical translation is limited by the need for personalisation and lingering efficacy concerns. Artificial intelligence (AI) has emerged as a transformative approach to advance the design, optimisation, and application of biomaterials in oral oncology. By integrating machine learning (ML) and data-driven modelling, AI enhances diagnostic accuracy through biosensing and radiomic analysis, guides the rational design of drug carriers and dosing regimens, and facilitates computer-aided scaffold fabrication for maxillofacial reconstruction. This review summarises recent advances at the intersection of AI and biomaterials in the context of oral cancer and OPMDs, highlighting innovations in early detection, targeted therapy, and postoperative repair. It also discusses current barriers, including data quality, model generalizability, and regulatory oversight, and outlines future directions for interdisciplinary research. When properly integrated, AI-enabled biomaterials hold considerable potential to deliver more precise, efficient, and patient-tailored solutions for oral cancer management.

Oral potentially malignant disorder (OPMD) remains a critical clinical challenge for cancer interception. However, the progression of OPMD toward oral squamous cell carcinoma (OSCC) is driven by profound cellular heterogeneity and dynamic microenvironmental remodeling. The mechanisms underlying these processes are not yet fully understood. Recent advancements in single-cell and spatial omics have facilitated high-resolution decoding of the precancerous landscape, unveiling multistep epithelial cell plasticity, fibroblast heterogeneity with extracellular matrix remodeling, immune suppression, and inflammatory reprogramming, as well as coupled metabolic and redox alterations that govern malignant transformation. These insights into the cellular mechanisms have led to a paradigm shift in the understanding of OPMD, reclassifying it as an ecosystem-level disease rather than a purely epithelial pathology. Nanomedicine is a potent platform for translating mechanistic knowledge into precision diagnostics and interventions at the precancerous stage. Nanomaterial-based strategies have been demonstrated to facilitate several critical processes, including enabling early lesion visualization and risk stratification, immune microenvironment reactivation, anti-fibrotic and anti-inflammatory remodeling, and targeted regulation of metabolic and oxidative stress pathways. A mounting body of evidence from preclinical and clinical studies lends support to the notion that nanotechnology-assisted early detection, microenvironmental reprogramming, and the interception of malignant transformation across oral and other precancerous conditions are indeed feasible. This review integrates single-cell-resolved mechanisms of OPMD progression with state-of-the-art nanomedicine-based diagnostic and therapeutic strategies, highlighting convergent biological axes and translational opportunities. By integrating single-cell biology with nanotechnology-driven precision medicine, this work is expected to improve the development of a nanomedicine framework for early cancer detection and treatment and outline future directions and challenges toward clinical implementation.

The combination of multikinase inhibitors with PD-1 blockade therapy has emerged as a promising strategy to overcome resistance to PD-1 blockade monotherapy across multiple cancer types, including gastric cancer. In this study, we report that the multikinase inhibitor lenvatinib selectively reduced the number of CD206+CD163+ immunosuppressive macrophages in the tumor microenvironment (TME) and increased antitumor immunity. Longitudinal immunoprofiling was conducted with paired (pre- and posttreatment) tumor samples from patients with advanced gastric cancer who received first- or second-line combination therapy with lenvatinib and pembrolizumab in the EPOC1706 clinical trial. Patients with abundant CD206+CD163+ immunosuppressive macrophage infiltration exhibited favorable responses to combination therapy, accompanied by a significant posttreatment reduction in these cells. Although this immunosuppressive macrophage infiltration was associated with resistance to PD-1 blockade monotherapy, it predicted a response to combination treatment: 8 of the 9 patients with >440 CD206+CD163+ immunosuppressive macrophages/mm2 responded, whereas none of the 8 patients who received monotherapy responded. Mechanistically, lenvatinib inhibited platelet-derived growth factor receptor α (PDGFRα)/fibroblast growth factor receptor (FGFR)-dependent p38 mitogen-activated protein kinase (MAPK) and AKT signaling pathways in F4/80highCD11bint immunosuppressive macrophages, triggering endoplasmic reticulum stress and an unresolved unfolded protein response, resulting in their apoptosis. Furthermore, in multiple animal models, the therapeutic efficacy of the combination was observed in tumors with abundant immunosuppressive macrophages with activated PDGFR/FGFR-AKT/p38 MAPK signaling. Therefore, we propose that the abundance of immunosuppressive macrophages in the TME could serve as a predictive biomarker for patient stratification to guide rational anti-PD-1-based combination therapy in gastric cancer, enabling mechanism-based combination cancer immunotherapy.

Gastric cancer with peritoneal metastasis has a poor prognosis and limited treatment options. Claudin 18.2 (CLDN18.2) CAR T therapy has shown activity in advanced gastric cancer, but its role in peritoneal disease remains insufficiently characterized. Here, we report three patients with gastric cancer and peritoneal metastasis from a phase I trial of satri-cel (NCT03874897), all with high CLDN18.2 expression. All three patients experienced clinical and radiologic benefit after treatment and achieved durable peritoneal disease control. After CLDN18.2-targeted CAR T-cell therapy, one patient underwent conversion surgery 8 months after infusion. After resection of subsequent bilateral ovarian metastases, no further disease progression was observed, corresponding to long-term control of peritoneal metastases. The other two patients achieved overall survival of 44 and 35 months, respectively. Although circulating CAR T-cells became nearly undetectable in peripheral blood by around day 30 after infusion, CAR T-cell infiltration was still detected in resected gastric or ovarian tumor tissues months later. These findings support substantial and durable activity of CLDN18.2 CAR T-cell therapy in gastric cancer with peritoneal metastasis.

Claudin 18.2 (CLDN18.2) is an attractive therapeutic target in gastric cancer (GC). Zolbetuximab, a CLDN18.2-targeted monoclonal antibody, has been approved in combination with chemotherapy as a first-line treatment option for patients with HER2-negative and CLDN18.2-positive gastric adenocarcinoma. This review summarizes current advances in CLDN18.2-targeted therapies for GC, with a focus on the clinical development of monoclonal antibodies (mAbs), particularly zolbetuximab and emerging next-generation antibodies, as well as mAb-based combination strategies. This review also summarizes CLDN18.2-targeted antibody-drug conjugates (ADCs), bispecific antibodies (BsAbs), and chimeric antigen receptor (CAR)-T cell therapy, highlighting emerging therapeutic strategies. Furthermore, major challenges and considerations in clinical and translational research are outlined. In conclusion, precision targeting of CLDN18.2 represents a promising approach in GC therapy, shifting from conventional chemotherapy to biomarker-guided strategies. Continued development of next-generation agents and rational combination therapies may enhance outcomes and expand benefit.

Perfluorooctanoic Acid (PFOA) has gained our attention for it is resistant to natural degradation and can migrate over long distances. We have already found that PFASs were related to gastric cancer in previous work. However, the systematic evidence on the gastric toxicity of PFOA and its mechanisms remains scarce. In this study, network toxicology pinpointed the peroxisome proliferator-activated receptor (PPAR) pathway as the pivotal driver between PFOA exposure and gastric cancer progression. In the cell experiments, PPAR inhibitor (10 μM) and ferroptosis inhibitor (2 μM) protect gastric cells from ferroptosis induced by PFOA (200 ng L-1). In the animal experiments, we set 3 PFOA concentrations (34.85 ng kg-1 day-1,1mg kg-1 day-1, 2.5 mg kg-1 day-1). Based on differential metabolic rate, the environmental dosage of PFOA in human was transferred to the lowest mice dosage. The RNA-seq analysis identified PPAR-γ as a differentially expressed gene. Compared with the normal group, the PFOA-L group of PPAR-γ decreased by 31.6%, the PFOA-M group of PPAR-γ decreased by 14.4%, and the PFOA-H group decreased by 46.8%. By organoid experiments, 5 days PFOA induced PPAR-γ downregulation and the increase of cancer marker P53. We elucidate that PFOA disrupts the PPAR-γ axis to induce gastric cancer progression, highlighting an overlooked gastric liability in PFOA health risk assessments.

The stomach is home to numerous nuclear receptor transcription factors (NRs) that can respond to food, toxins, and other ingested agents. Conversely, signals secreted from other organs (e.g., hormones) can engage gastric NRs to modulate gastric physiology. Thus, there is a rich potential interface between external and internal signals that gastric NRs might respond to and interpret. Here, we seek to comprehensively review the role of NRs in gastric homeostasis and disease pathogenesis. NRs are evolutionarily conserved proteins that regulate gene transcription by interpreting hormonal and environmental signals. We explore NR roles in normal stomach development, cell fate determination, and responses to dietary compounds and xenobiotics. The last topic is of particular recent importance 1) because NRs stimulated by ingested agents might directly regulate gastric physiology like the relative activity of acid-secreting and stem cells and 2) because the stomach is one of the first organs to encounter dietary compounds and pollutants. Additionally, we review the emerging yet understudied field of gastro-endocrinology, exploring how systemic endocrine circuits influence the stomach's function. We also discuss how NRs contribute to pathological conditions like precancerous lesions and cancer. Additionally, we summarize known agonists, antagonists, and coregulatory proteins, highlighting potential therapeutic targets. Understanding NR roles could pave the way for a better understanding of dietary and environmental toxin exposure and also lead to innovative treatments for gastric disorders, including gastritis, gastric intestinal metaplasia, and gastric cancer.

Exposed constantly to environmental challenges, the stomach undergoes highly reversible cycles of regeneration and recovery. Although abnormal activation of regeneration is known to be associated with cancer, the mechanisms underlying tissue restoration remain unclear. Our single-cell gene expression and chromatin analysis defined cell state dynamics during regeneration and recovery, identifying the Brahma-related gene 1(BRG1)/BRM-associated factor (BAF) chromatin remodeling complex during recovery. Strikingly, deletion of AT-rich interaction domain 1A (Arid1a), a subunit of the BAF complex and the second most frequently mutated gene in gastric cancer, impaired recovery across multiple murine injury models, resulting in a persistent regenerative state. Integrative analyses combining single-cell multiome and chromatin immunoprecipitation sequencing (ChIP-seq) demonstrated that the BAF complex recruits lineage-specific transcription factors such as MIST1 and estrogen related receptor gamma (ERRγ) to regulate enhancers of recovery genes. Notably, deletion of Trp53 in the unresolved regenerative state caused by Arid1a loss is sufficient to drive cancer development and invasion, revealing the epigenetic mechanisms bridging gastric regeneration, recovery, and cancer.

Systemic chemotherapy for gastric cancer is frequently compromised by debilitating, dose-dependent toxicities, necessitating innovative adjunctive strategies that balance therapeutic efficacy with systemic safety. While natural products represent a promising source of candidates, their clinical translation is frequently hindered by intrinsic pharmacokinetic barriers. To address this, we developed the oral resident binary intestinal therapy (ORBIT) system, an excipient-free nanocapsule platform engineered for prolonged gut residency. Unlike conventional pharmaceutical formulations that depend on synthetic excipients, ORBIT is an excipient-free, self-assembled nanocapsule composed of a high-density dopamine-functionalized hyaluronic acid (hDAHA) shell encapsulating a berberine (BBR) core. This unique architecture provides a protective shield for BBR against premature degradation while utilizing the adhesive properties of the hDAHA shell to enhance gastrointestinal retention. ORBIT enables sustained, pH-responsive drug release, providing robust in situ shielding of the intestinal mucosa against oxaliplatin-induced injury and preserving epithelial barrier integrity. In preclinical gastric cancer models, the ORBIT regimen synergistically potentiates oxaliplatin-mediated tumor suppression while dramatically ameliorating systemic toxicity. Mechanistically, ORBIT administration profoundly remodels the microbiome-immune axis, specifically enriching beneficial commensals such as Akkermansia to drive increased intratumoral infiltration of CD8⁺ T cells. This study establishes a new oral nanotherapeutic paradigm for chemo-immunotherapy, presenting a compelling and clinically translatable strategy to optimize gastric cancer treatment outcomes.

Clear cell renal cell carcinoma (ccRCC) is largely driven by the transcription factor hypoxia-inducible factor 2α (HIF-2α)1. Here we show that monotherapy with casdatifan-an orally bioavailable, potent and selective HIF-2α inhibitor2-produces meaningful, durable antitumour activity with manageable safety in individuals with refractory metastatic ccRCC. Dose-expansion data from the ARC-20 study ( NCT05536141 ) are presented, including for the 100 mg once daily (QD) cohort (n = 32) and the total cohort (n = 127). Treatment discontinuation from casdatifan-related adverse events was infrequent (3%), and class-effect toxicities included anaemia and hypoxia. The confirmed objective response rates (ORRs) were 35% (95% confidence intervals (CI) = 19-55%; 100 mg QD) and 31% (95% CI = 23-40%; total); median progression-free survival (PFS) was not estimable (95% CI = 5.7-not estimable; 100 mg QD) and 12.2 months (9.4-20.6; total). Greater maximal reductions in serum erythropoietin were associated with improved clinical outcomes, including a higher ORR (P = 0.001), lower rates of progressive disease (P = 0.003) and longer PFS (P = 0.006). Erythropoietin expression was restricted to cancer cells and was significantly higher at the mRNA level in patients with clinical benefit. Concordantly, HIF-2α protein expression and HIF-2α expression signature were associated with prolonged PFS. Overall, our findings show that casdatifan achieves meaningful, durable responses with manageable safety. These data establish a link between on-target HIF-2α pathway modulation, tumour biology and clinical efficacy.

Immune checkpoint inhibitor-based combinations represent the standard of care for advanced renal cell carcinoma. The OMNIVORE trial investigated a response-adaptive strategy, including treatment discontinuation in early nivolumab responders (Arm A) and salvage ipilimumab addition in non-responders (Arm B). Here we report outcomes with extended follow-up. OMNIVORE was a phase II response-adaptive trial in which patients received induction nivolumab monotherapy. Patients achieving a confirmed complete or partial response discontinued nivolumab and entered observation (Arm A), while patients with stable disease or progressive disease received two doses of ipilimumab added to ongoing nivolumab (Arm B). This analysis characterizes long-term overall survival across the full study cohort and durability of response among patients who discontinued nivolumab following an early objective response. Of 83 patients who initiated treatment, 12 (14%) were allocated to Arm A and 57 (69%) to Arm B, with a median follow-up among living patients of 59.4 months (range 29.1-85.4 months) in Arm A and 31.4 months (range 4.6-62.6 months) in Arm B. The 3-year overall survival rate from nivolumab initiation was 64% (95% CI 51% to 74%) in the overall cohort, 83% (95% CI 48% to 96%) in Arm A, and 63% (95% CI 47% to 76%) in Arm B. Of the 12 Arm A patients, 6 (50%) remained off nivolumab at 1 year following treatment discontinuation, of whom 5 maintained responses beyond 43 months off therapy, with all remaining alive at last known follow-up, with overall survival ranging from 48.8 to 85.4 months from nivolumab initiation. Of the six patients who resumed treatment, only one achieved a durable complete response remaining on treatment at 83.2 months from nivolumab initiation. All 57 Arm B patients discontinued treatment with a median treatment duration of 3.7 months (range 1-24.8 months) and a median progression-free survival from nivolumab plus ipilimumab initiation of 4.6 months (95% CI 2.7 to 6.5 months). With extended follow-up, a meaningful subset of patients achieving an early objective response to nivolumab maintained prolonged treatment-free survival following a short course of treatment. Salvage ipilimumab in nivolumab non-responders demonstrated limited benefit, reinforcing that upfront concurrent dual checkpoint blockade remains the preferred approach. NCT03203473.

Radiotherapy-based metastasis-directed therapy (MDT) has emerged as a treatment strategy for oligometastatic clear cell renal cell carcinoma (ccRCC). However, optimal integration of MDT with immune checkpoint inhibition (ICI) is unclear, especially in light of the M1 no evidence of disease subgroup analysis of KEYNOTE-564. We undertook an exploratory cohort study of two previously reported trials evaluating either MDT+ICI (NCT02855203) or MDT (NCT03575611) for oligometastatic ccRCC. The objective was primarily to compare RECIST-defined progression-free survival (PFS) and secondarily to compare peripheral immune populations. Among the 150 patients included in the analysis (MDT+ICI: 30; MDT: 120), the MDT+ICI cohort had more metastases (median 3 vs 1) and was slightly younger (median 62 vs 66) than MDT. After a median follow-up time of 34 mo, there was evidence for longer PFS after MDT+ICI vs MDT that did not reach statistical significance (hazard ratio, 0.57; 95% confidence interval: 0.32-1.02; p = 0.058). Interaction testing demonstrated greater PFS benefit with MDT+ICI among patients with previous receipt of systemic therapy. Immediate systemic induction of activated CD8+ T cells (ICOS+) was more common after MDT+ICI, as were decreases in less functional CD8+ T cell subsets. Taken together, this study provides evidence that adding maintenance ICI to MDT improves clinical outcomes for patients with oligometastatic ccRCC. MDT+ICI-evoked immunomodulatory signals are promising and support the observed superiority in clinical outcomes. The A Randomized Trial of Maintenance Systemic Therapy After Radiation for Oligometastatic Renal Cell Carcinoma (ASTROs) trial (NCT06004336) has been initiated to test the hypotheses generated by the present study.

Screening for kidney cancer has been identified as a research priority by several independent initiatives. Previous work revealed that the relatively low prevalence of kidney cancer in untargeted asymptomatic individuals hinders the cost-effectiveness and clinical utility of population screening. We, therefore, undertake a comprehensive literature review summarizing novel screening concepts beyond whole population screening, including targeting high-risk populations (to increase disease prevalence), screening for kidney cancer in combination with other abdominal conditions (to reduce costs), or combining both of these approaches. Risk-stratified screening may be undertaken using phenotypic and/or genetic risk scores. Phenotypic risk scores for kidney cancer generally achieve reasonable discrimination (area under the receiver operating characteristic curve between 0.67 and 0.71 in a mixed sex cohort), with the best performing model being based on age, sex, body mass index, smoking status, hypertension diagnosis, and systolic and diastolic blood pressure. Adding genetic to phenotypic data is costly and provides only a negligible gain in discrimination (an increase in the area under the receiver operating characteristic curve of 0.007 from 0.716 to 0.723), meaning it is unlikely to represent a clinically useful strategy. Alternatively, screening for kidney cancer may be combined with other conditions to maximize efficiency, including multicancer early detection tests and combined screening using ultrasonography (concurrently with abdominal aortic aneurysm screening) or abdominal computed tomography (concurrently with lung cancer screening). The latter strategy is most promising and has the highest public acceptability. In summary, we highlight benefits and harms of different screening strategies beyond traditional population screening and offer a comprehensive and contemporary summary of recent advances in this field and future research avenues.

Clear cell renal cell carcinoma (ccRCC) is the most common and aggressive subtype of kidney cancer. Loss of von Hippel-Lindau (VHL) and the consequent activation of hypoxia-inducible factor-α (HIFα, especially HIF2α) plays an essential role in ccRCC initiation and progression. The VHL-HIF2α axis as the main driver for ccRCC may present specific opportunities to control the disease by cotargeting HIF2α with belzutifan and another vulnerability. This study elucidates the synthetic essentiality of TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) in VHL-deficient ccRCC. Upregulated in ccRCC in a VHL-HIF2α-dependent manner, TRAIL is selectively essential in ccRCC cells, promoting cell proliferation by activating the p38 MAPK pathway and facilitating G1/S phase transition. Depletion of endogenous TRAIL or inhibition of HIF2α with belzutifan sensitizes ccRCC cell and tumor models to recombinant TRAIL, presenting a promising avenue for combination therapy in ccRCC.

Sarcomatoid renal cell carcinoma (sRCC), a lethal variant arising through sarcomatoid dedifferentiation of epithelioid RCC (eRCC), poses significant clinical challenges due to the lack of molecular biomarkers and limited therapeutic options, with a median survival time for patients of less than 12 months. To dissect the molecular basis of sarcomatoid dedifferentiation and develop novel therapeutic strategies for sRCC. Guided by the coprogenitor theory and tumor plasticity principles, we developed a differentiation-induction strategy targeting sarcomatoid dedifferentiation. Integrated transcriptomic and proteomic profiling revealed PLOD2 as a candidate therapeutic target. Spatial profiling of clinical sRCC specimens revealed selective PLOD2 overexpression in sarcomatoid components. Functional assays, including genetic ablation and pharmacological inhibition, were performed in sRCC cell lines and xenograft models to validate the role of PLOD2 in conferring sarcomatoid dedifferentiation plasticity and sarcomatoid morphology. Therapeutic significance was evaluated by PLOD2 intervention alone or in combination with conventional therapies (doxorubicin, gemcitabine, IFN-α, and axitinib). Compared with neighboring epithelioid RCC and adjacent normal components, PLOD2 expression was markedly upregulated in sarcomatoid regions. PLOD2 fuelled sarcomatoid dedifferentiation plasticity by activating cancer stemness, dedifferentiation, and EMT, partially via downstream activation of DCLK1, a cancer stem cell marker. PLOD2 ablation reversed sarcomatoid phenotypes, restored epithelioid differentiation, and sensitized tumors to conventional RCC therapies. Notably, minoxidil, an FDA-approved PLOD2 inhibitor, effectively suppressed sarcomatoid features and synergized with standard treatments in preclinical models. Targeting PLOD2-mediated tumor plasticity represents a novel differentiation-inducing strategy to reprogram sRCC into therapy-responsive epithelioid states. The repurposing potential of minoxidil provides an immediate translatable strategy to improve clinical outcomes in this treatment-resistant malignancy.

Accurate and comparable human papillomavirus (HPV) testing is essential for vaccine research, surveillance, and cervical cancer screening. Since 2008, the WHO HPV Laboratory Network has conducted global HPV proficiency testing traceable to International Standards. Here, we present a retrospective analysis of global HPV genotyping and screening proficiency panel results collected between 2008 and 2024. The HPV genotyping panel consists of 43 blinded samples including HPV6/11, all oncogenic and vaccine-targeted HPV types. A screening panel, launched in 2022, includes 13 blinded samples for cervical screening. Laboratories worldwide test panels using their methods, and results are evaluated at the International HPV Reference Center. Proficiency is defined as absence of false positives and detection of HPV16/18 at 10 IU/µL, and other oncogenic types at 100 IU/µL (genotyping) or 1000 IU/µL (screening). Genotyping panel participation increased from 54 laboratories (2008) to > 130 (2021). Proficiency rose from ~25% to > 80% by 2024, with > 99% correct detection for most genotypes. Optional low-copy challenges (1 IU/µL HPV16/18) were detected by > 95% of laboratories by 2024. Screening panel participation increased from 84 laboratories (2022) to 132 (2024), with proficiency improving from 77% to 95% and > 96% datasets free of false positives. Annual HPV proficiency testing provides global laboratory quality assurance supporting cervical cancer elimination.

Human papillomavirus (HPV) infection is a leading cause of cervical cancer and other malignancies, underscoring the urgent need for accurate and rapid early diagnostic strategies. Herein, we report a dual-mode colorimetric and electron spin resonance (ESR) method for the qualitative detection of HPV-16, based on the integration of the CRISPR/Cas12a system with a metal-organic framework (MOF). A peroxidase-mimicking iron-based MOF, designated MIL-101(Fe), was conjugated to magnetic beads via a single-stranded DNA linker to serve as a signal probe. Upon recognition of the target nucleic acid, MIL-101(Fe) catalyzes the decomposition of hydrogen peroxide to generate hydroxyl radicals, which oxidize a chromogenic substrate to produce a visible color change. Meanwhile, the generated radicals are captured by a spin trap and detected by ESR spectroscopy. The assay enables sensitive and rapid detection of HPV-16, with clear discrimination even in mixtures containing both HPV-16 and HPV-18. Importantly, when evaluated with clinical specimens, the method achieved 100% sensitivity and specificity. Overall, this work provides a feasible and promising strategy for ultrasensitive nucleic acid detection and offers a new avenue for advancing CRISPR-based multimodal diagnostic platforms toward practical applications.

High-risk human papillomaviruses (HPV) are the main etiological agents of cervical cancer. The viral early promoter regulates the expression of E6/E7 oncoproteins, and its transcriptional activity is positively or negatively regulated by host transcription factors (TFs) that are able to bind to the viral long control region (LCR). In this study, we assessed the impact of a TF library on the early transcriptional activity of high-risk HPV-16 and -18, and among these TF, we selected and investigated the impact of GATA3, as well as its potential role as a prognostic biomarker for the development of cervical cancer. Luciferase reporter assays demonstrated that GATA3 negatively influences the transcriptional activity of HPV-16 and -18, downregulating E6 and E7 mRNA levels, with in silico and in vivo assays indicating that this effect is due to GATA3 direct binding to the viral LCR. Subsequently, we evaluated GATA3 levels in normal and HPV-immortalized epithelial raft cultures and cervical cancer samples by immunohistochemistry, also accessing the TF presence in pre-neoplastic intraepithelial cervical lesions (CIN) by immunofluorescence assays, further correlating GATA3 protein expression with the presence of HPV E6/E7 mRNA. This approach revealed an apparent inverse correlation between GATA3 expression and the grade of CIN lesions, as well as with the presence of viral oncogene transcripts. When accessing GATA3 expression in cancer samples, we observed a significant correlation between the absence of GATA3 and higher stages of cancer. Thus, our data indicates that the loss of GATA3 expression contributes to high-risk HPV-mediated cervical carcinogenesis, with the TF possibility acting as a protective factor whose absence enables sustained viral oncogene expression and disease progression in the cervical tissue.

Oncogenic viruses cause approximately 20% of cancers globally burden, with Epstein-Barr virus and high-risk Human papillomavirus recognized as major contributors to epithelial malignancies. Increasing evidence suggests that EBV-HPV co-infection may enhance tumour progression through overlapping molecular and immunological mechanisms, particularly in cervical, oropharyngeal, and nasopharyngeal cancers. This review critically summarizes current evidence regarding the cooperative role of EBV and HPV in carcinogenesis while distinguishing viral co-presence from biologically active co-infection. EBV latent proteins, including LMP1, LMP2A, and EBNA1, activate oncogenic signalling pathways such as NF-κB, PI3K/Akt, and JAK/STAT, whereas HPV oncoproteins E6 and E7 disrupt p53 and retinoblastoma (Rb) tumour suppressor pathways. Together, these alterations may promote genomic instability, chronic inflammation, immune evasion, epigenetic dysregulation, and epithelial-mesenchymal transition (EMT), thereby enhancing invasive and metastatic potential. Epidemiological studies report higher frequencies of EBV-HPV co-detection in advanced lesions and aggressive tumours; however, causal synergy remains insufficiently validated because of methodological heterogeneity and variability in viral detection techniques, including PCR, in situ hybridization, and immunohistochemistry. Emerging technologies such as spatial transcriptomics and single-cell profiling may improve characterization of biologically meaningful co-infection. In addition, circulating viral DNA, viral microRNAs, and HPV genotyping are being explored as biomarkers for disease monitoring and prognosis. Therapeutic strategies targeting viral oncogenes, immune checkpoints, and gene-editing technologies also represent promising investigational approaches. Overall, EBV-HPV co-infection represents a biologically plausible but incompletely understood contributor to tumour aggressiveness, emphasizing the need for standardized diagnostics, longitudinal studies, and functional experimental models.

Photodynamic therapy (PDT) is effective for localized cancers, but its reliance on external light limits treatment depth and uniformity. Herein, we report a programmable supramolecular chemiluminescence (CL)-induced PDT platform that generates reactive oxygen species (ROS) in the absence of light irradiation and displays intrinsic cancer selectivity. This platform is constructed through the supramolecular interaction between enzyme-responsive chemiluminescent spiroadamantyl phenoxy-1,2-dioxetane probes (ADOP, ADOE, and ADOG) and luminescent iridium(III) complexes bearing a trimethyl-β-cyclodextrin (TMCD) host [Ir(N∧C)2(bpy-TMCD)](Cl) (HN∧C = 2-phenylpyridine (Hppy) (1a), 2-phenylquinoline (Hpq) (2a), and 2-(1-naphthyl)benzothiazole (Hbsn) (3a)). The chemiluminescent probes exhibited CL upon reaction with alkaline phosphatase (ALP), porcine liver esterase (PLE), and β-galactosidase (β-gal); while the iridium(III) complexes showed intense phosphorescence with efficient singlet oxygen production, distinct intracellular localization, and tunable (photo)cytotoxicity. In aqueous solutions, mixing the chemiluminescent probes with TMCD-tagged iridium(III) complexes afforded supramolecular adducts that, upon enzyme activation, underwent chemiluminescence resonance energy transfer (CRET) from the dioxetane donors to the iridium(III) acceptors. Remarkably, the ALP-responsive conjugate formed from probe ADOP and complex 3a (adduct ADOP-3a) selectively generated ROS and elicited potent cytotoxicity in cancerous HeLa cells and spheroids under light-free conditions, with apoptosis as the predominant cell death pathway, while remaining noncytotoxic in normal HEK293 cells. This modular platform couples supramolecular host-guest assembly, enzyme-specific activation, and CRET-induced ROS generation to overcome the limitations of conventional PDT, enabling cancer-selective, light-free therapy.

Cervical cancer remains a leading cause of cancer-related mortality among women worldwide, with mortality disproportionately concentrated in low-and middle-income countries (LMICs), where screening infrastructure is limited. Although persistent high-risk human papillomavirus (hrHPV) infection drives nearly all cervical cancers through a prolonged preinvasive window, making the disease both preventable and detectable, current screening modalities face biological and operational limitations that constrain their global impact. Cytology-based methods suffer from moderate sensitivity and subjective interpretations. HPV DNA testing is highly sensitive, it lacks specificity for transforming infections and shifts the diagnostic burden toward triage and colposcopy. Meanwhile, visual inspection methods are accessible, they offer limited reproducibility. In this review, we adopt a target-centric diagnostic framework that organizes cervical cancer screening not by detection platform but by the biological class of the marker being interrogated. We first examined the molecular pathogenesis of cervical carcinogenesis, including HPV genotype-specific biology, viral integration dynamics, oncogene-driven transformation, and epigenetic consolidation, to establish the biological rationale for each biomarker category. We then systematically evaluated conventional screening modalities and their limitations before reviewing emerging diagnostic technologies across four target domains: HPV-derived markers (DNA, mRNA, capsid proteins), host cell-cycle regulators (p16INK4a, Ki-67), non-protein biomarkers (biothiols, volatile metabolites), and marker-free platforms (Raman/FT-IR spectroscopy, AI-assisted cytopathology). Particular emphasis is placed on point-of-care (POC) deployability, noninvasive sampling strategies, and the integration of multilayered risk stratification into scalable screening pathways. By mapping diagnostic innovation onto the biological continuum of cervical carcinogenesis, this review provides a conceptual foundation for developing next-generation screening approaches that align molecular precision with global accessibility, thereby supporting progress toward the World Health Organization goal of cervical cancer elimination.

MicroRNA-21 (miR-21) is a key bladder cancer diagnostic biomarker, but its ultra-low abundance and matrix interference in clinical samples impede accurate detection. Herein, we developed a sensitive electrochemical biosensor integrating CHA-HCR cascade amplification with oxygen vacancy (OVac)-rich ultra-thin core-shell Au@MnO2 nanozymes for clinical miR-21 detection. The Au@MnO2 exhibited excellent oxidase-mimicking activity: δ-MnO2 shell OVac form efficient electron transfer pathways (reducing environmental sensitivity), while the Au core accelerates electron transfer, synergistically enhancing signals. miR-21 triggers CHA amplification; its products induce HCR to form long-chain DNA that captured by nanozymes, achieving efficient signal amplification via the masked active sites. Under optimal conditions, the biosensor establishes a detection range of 10 fM-10 nM (LOD: 1.32 fM) with superior selectivity and stability. Notably, it accurately detects miR-21 in urine samples, with results highly consistent with PCR (nucleic acid detection gold standard), confirming clinical practicality. This biosensor provides a reliable supplementary platform for bladder cancer diagnosis.

The PURE-01 trial evaluated three cycles of pembrolizumab 200 mg followed by radical cystectomy (RC) in patients with cT2-3b N0 M0 muscle-invasive bladder cancer (MIBC) ineligible for or refusing cisplatin. A total of 155 patients were treated. We report survival outcomes after median follow-up of >60 mo and associations with pathological response rates and biomarker levels. In the intention-to-treat population, the 5-yr event-free survival rate was 68% and the 5-yr overall survival (OS) rate was 77%. In the RC cohort, the 5-yr recurrence-free survival (RFS) rate was 77%. Pathological response categories were significantly associated with survival, with a 5-yr OS rate of ∼90% in the group achieving a complete or major response (p < 0.001). The 5-yr cumulative risk of recurrence was 19%, with three of 31 relapses occurring beyond 5 yr after RC. Eight patients refused RC and underwent transurethral resection; seven remain alive and disease-free. Transcriptome-wide profiling of pretreatment tissue was available for 102 patients. Stratification by genomic subtyping classifier showed that the claudin-low subtype (n = 14) had the highest 5-yr RFS rate, with a 5-yr OS rate of 93%. At long-term follow-up, PURE-01 demonstrates sustained survival outcomes and confirms the prognostic relevance of pathological response and molecular subtypes. These results support further investigation of single-agent immunotherapy as a potential de-escalation strategy in selected patients.

Rapid and multiplexed detection of urinary microRNAs (miRNAs) is crucial for the noninvasive diagnosis and monitoring of bladder cancer. In this study, we develop a dual-channel fluorescence platform based on filler-mediated quantum dot-DNA nanospheres for simultaneous detection of miRNA-21 and miRNA-96. The nanospheres are constructed via a streptavidin-biotin cross-linked bidirectional hybridization chain reaction and compacted by phenyl-lactic acid, enabling high-density encapsulation of distinct QDs. Upon target miRNA recognition, a strand-displacement cascade triggers nanosphere disassembly and QDs release, restoring well-resolved fluorescence at 500 and 624 nm. This enzyme-free, extraction-free assay is completed in 40 min. It achieves attomolar-level sensitivity, with detection limits of 30 aM for miRNA-21 and 40 aM for miRNA-96, while maintaining high selectivity against mismatched and nontarget miRNAs. Clinical validation using urine samples from 45 patients with bladder cancer, 22 patients with other urological diseases, and 20 healthy controls indicated strong concordance with quantitative polymerase chain reaction. Integrated dual-miRNA analysis significantly enhanced diagnostic performance, yielding 91.0% sensitivity, 80.0% specificity, and an area under the curve of 0.91. The platform provides a rapid tool for noninvasive miRNA profiling, with potential to distinguish bladder cancer from healthy individuals and other urological diseases, thereby improving bladder-cancer screening, stratification, and recurrence monitoring.

Sonodynamic therapy (SDT) represents a promising methodology that employs sonosensitizers in conjunction with low-intensity ultrasound for the eradication of malignant tumors, featuring precise treatment capabilities, deep tissue penetrability, and minimal side effects. Conventional sonosensitizers often face challenges such as aggregation-caused quenching (ACQ), which hampers the efficiency of reactive oxygen species (ROS) generation. In this study, we report a novel benzothiadiazole-based sonosensitizer derivative, BBTPA, exhibiting aggregation-induced emission (AIE) characteristics. By co-assembling BBTPA with the ROS-responsive polymer PMD and the amphiphilic polymer DSPE-PEG2000, we engineered BBTPA nanoparticles (NPBBTPA). Upon ultrasound exposure, NPBBTPA produces ROS efficiently, inducing mitochondrial damage and triggering pyroptotic cell death. Moreover, NPBBTPA induces immunogenic cell death (ICD) under ultrasound stimulation, thus enhancing antitumor immune responses. This study extends the utility of AIE-based sonodynamic agents in efficient cancer therapy, holding promising prospects for bladder cancer treatment.

Peptide-drug conjugates (PDC) have recently garnered substantial attention as promising strategies for targeted tumor therapy. However, the clinical efficacy of PDC is limited by poor membrane permeability and inherent lysosomal sequestration. Herein, we report an in-situ cascade assembled peptide-drug conjugate (ISCA-PDC) accomplished by integrating CXCR4-targeting cyclic peptide (Cyclo(DTyr-NMe-DOrn-Arg-2Nal-Gly)), a lysosomal-triggered assembled peptide linker (VEALYL) decorated with pH-sensitive moiety (cis-aconitic anhydride, CAA), and cytotoxic payload (camptothecin, CPT), to improve the tumor membrane permeability and realize lysosomal destabilization, ultimately enhancing the chemotherapy of bladder cancer. In the acidic microenvironment (pH 6.5) of the tumor, ISCA-PDC could first self-assemble into nanoparticles (NPs-PDC) after the hydrolysis of CAA and quickly enter the lysosome via CXCR4-mediated endocytosis to improve tumor membrane permeability. Following transformation into nanofibers (NFs-PDC) within lysosomes (pH 5.0), the permeability of the lysosomes was markedly enhanced, resulting in cathepsin B-induced apoptosis and CPT release. In addition, ISCA-PDC exhibited highly potent antitumor efficacy, which extended the overall survival of tumor recurrence model mice and led to the eradication and regression of T24-luc orthotopic xenograft mice. The concept of in-situ cascade-assembled PDC can be extended by conjugation with other chemotherapeutic agents, suggesting a generalizable strategy for nanotherapeutic enhancement in solid tumors.

Esophageal squamous cell carcinoma (ESCC) exhibits heterogeneous responses to chemoimmunotherapy, with only a minority achieving durable benefit, necessitating dynamic precision monitoring. Through longitudinal plasma metabolomics of 541 serial samples from 252 ESCORT-1st trial patients receiving chemoimmunotherapy plus 3 independent cohorts of 288 samples, we established an integrated risk assessment framework spanning the entire therapeutic continuum: (i) a baseline predictor for initial responders based on metabolite signatures; (ii) an on-treatment predictor in prognosticating long-term responders among initial ones based on treatment-induced metabolic shift patterns; and (iii) a real-time model based on dual alteration of sphingolipid and glycerophospholipid dynamically stratifying progression risk. Meanwhile, 2 dietary metabolites, garlic-derived S-allyl-L-cysteine and cruciferous vegetable-derived indole-3-carbinol, were confirmed to improve outcomes by promoting NK-cell infiltration and reversing CD8+ T-cell exhaustion. In conclusion, we provide the first metabolomic roadmap for precision chemoimmunotherapy in ESCC, unifying baseline prediction, longitudinal surveillance, and dietary modulation into a clinically actionable paradigm. We established a large-scale plasma metabolomic database from patients with ESCC undergoing chemoimmunotherapy, defining the first noninvasive, comprehensive, and precise monitoring framework for treatment response prediction and risk assessment. Our findings reveal clinically actionable dietary metabolites that may serve as readily accessible adjuvants to enhance chemoimmunotherapy efficacy.

Neoadjuvant immunotherapy improves outcomes in esophageal squamous cell carcinoma (ESCC), yet ∼70% of patients fail to respond. Pretreatment biopsies and plasma provide critical opportunities for biomarker discovery. In this study, we performed plasma proteomic profiling and identified soluble glycoprotein nonmetastatic melanoma protein B (sGPNMB) as the most elevated circulating protein in nonresponders. Mechanistically, tumor cell-derived sGPNMB suppressed CD8+ T-cell receptor signaling via the SDC4-CD148 axis to induce functional exhaustion, with secretion being required for its immunosuppressive activity. Cancer-associated fibroblast-epithelial (CAF-Epi) niches promoted SOX2 upregulation in tumor cells, transcriptionally activating GPNMB expression. In humanized patient-derived xenograft models, circulating GPNMB levels predicted response to PD-1 blockade, and GPNMB inhibition synergized with therapy. Across retrospective cohorts and a prospective clinical trial, a multimodal model combining plasma GPNMB levels, CAF-Epi niche detection, and clinical-pathologic features achieved robust predictive accuracy for immunotherapy response and survival. These findings establish a spatial-circulating biomarker framework for precision ESCC immunotherapy. Tumor-derived soluble GPNMB, transcriptionally activated by SOX2 within CAF-Epi niches, drives CD8+ T-cell exhaustion and resistance to PD-1 blockade in ESCC. Integrating circulating GPNMB levels with CAF-Epi niche features and clinical-pathologic factors, we develop and validate a clinically scalable multimodal model for predicting immunotherapy response.

The facultative intracellular bacterium Fusobacterium nucleatum (Fn) promotes tumorigenesis and progression in esophageal squamous cell carcinoma (ESCC). The intracellular survival strategy of Fn and whether Fn can spread through cell‒cell contact in intratumoral tissues and, if so, the underlying mechanisms and implications are currently unknown. Here, we report that Fn accumulates in macrophages from ESCC tumors and paracancerous normal tissues. We further revealed that Fn-induced macrophage mitophagy through the PINK1-Parkin-independent pathway decreases excessive mitochondrial ROS production to promote survival. Furthermore, Fn drives a biphasic metabolic switch between glycolysis and oxidative phosphorylation in macrophages to support the bioenergetic demands of survival. Notably, Fn can be carried by macrophages to tumor sites, where it promotes tumor metastasis via the CCL2-CCR2 axis in ESCC. Treatment with a mitochondrial division inhibitor (mdivi-1) reduced the intracellular Fn concentration and inhibited Fn-positive tumor metastasis in mice. This study highlights the crucial interactions between Fn and host macrophages that influence tumor progression. These findings indicate that mitophagy inhibitors or mitophagy machinery targeting may serve as efficient therapeutic strategies to treat Fn-positive tumors.

Osteosarcoma (OSA) is the most common primary malignant bone tumor in children and adolescents, while no further improvement in survival has been achieved in the past few decades. Microwave ablation (MWA) is an emerging thermal therapy to ablate malignant bone tumors that induces antitumor immunity, while intrinsic mechanisms remain elusive. This study uncovered that MWA treatment promoted interleukin (IL)-18 expression and enhanced the dendritic cell (DC) functions, as well as CD8+ T cells infiltration. Combining IL-18 and anti-programmed cell death protein 1 (PD-1) displayed the additive tumor suppression effects. MWA treated OSA was co-cultured with bone marrow dendritic cells (BMDC) in vitro. RNA-sequencing was performed to analyze the intrinsic mechanism of BMDC after stimulated with MWA treated OSA. Mouse model of primary osteosarcoma of the femur was established for in vivo immune response evaluation, and this model was used to measure the additive tumor eradication of IL-18 and anti-PD-1 combination therapy. MWA promoted the activation, antigen uptake and antigen cross-presentation of BMDCs. Mouse models demonstrated that MWA treatment could elevate the proportion of CD8+ T cells and CD11c+DCs both in tumors and draining lymph nodes. The tumor suppression was mediated through the release of IL-18 by MWA-treated OSA and interaction with the IL-18 receptor on DCs, which was reversed after anti-IL-18 antibodies application. Besides, IL-18 cytokine exerted the tumor eradication effect and showed the additive role against OSA when combining anti-PD-1 therapy. Our study demonstrated a critical role of IL-18-mediated signaling in the induction of protective immune responses against OSA, which provided a novel mechanism to the explanation of MWA-induced immune activation, and IL-18 cytokine treatment could be considered a feasible, effective therapeutic approach for OSA in the clinic.

Sarcomas are rare, diverse malignancies with limited therapeutic options and poor clinical outcomes. Preclinical models that preserve tumour biology are urgently needed to advance mechanistic understanding and functional precision oncology. We established and comprehensively characterized 29 early-passage patient-derived sarcoma cell (PDC) cultures from 19 patients, representing 11 sarcoma subtypes. Multi-region and multi-site sampling enabled generation of PDCs from spatially distinct areas of individual tumours and from matched primary, recurrent and metastatic lesions. PDCs underwent genomic, transcriptomic and proteomic profiling alongside extracellular vesicle (EV) biomarker evaluation and phenotypic and drug-response assays. Copy-number variant (CNV) analysis revealed recurrent alterations affecting key regulators of cell cycle control and growth signalling. Bulk RNA-sequencing captured substantial inter- and intra-subtype heterogeneity. Proteomic and EV analyses recapitulated subtype- and site-specific differences. A functional drug screen of 38 clinically relevant and investigational agents identified both shared and divergent therapeutic vulnerabilities in the different PDCs. These results demonstrate that early-passage sarcoma PDCs can be regarded as biologically faithful and experimentally tractable models that capture lineage identity, tumour evolution and functional heterogeneity. Integrated multi-omic and functional profiling reveals therapeutic vulnerabilities not evident from genomic data only, supporting PDCs as valuable platforms for translational sarcoma research. A PDC biobank that addresses a critical gap in sarcoma research. Investigation of intra-patient and inter-patient heterogeneity. Comprehensive characterization shows truthful recapitulation of sarcoma features. Functional assays highlight the importance of functional models in cancer research.

Fertility-preserving treatments for young women with endometrioid endometrial carcinoma (EEC) have limited effectiveness, with only ∼50% of patients responding to first-line progesterone therapy. The absence of reliable methods to predict clinical response during the six-month treatment window creates substantial clinical uncertainty. Here, we developed EEC patient-derived tumor-like cell clusters (PTCs). Utilizing label-free hyperspectral stimulated Raman scattering microscopy in PTCs, we identified cholesteryl ester as an independent biomarker for progesterone response, achieving 86.2% predictive accuracy in a retrospective clinical cohort. Strikingly, progesterone-insensitive patients receiving progesterone-statin combination therapy exhibited a significantly higher 6-month complete response rate compared to those treated with progesterone alone (66.67% vs. 7.69%; HR = 13.00, 95% confidence interval (CI): 2.80-60.28, P < 0.001), including one documented case of post-treatment conception culminating in live birth. Mechanistically, fibroblast-specific cholesteryl ester accumulation was strongly associated with progesterone insensitivity. These results present a biomarker-guided strategy to optimize personalized fertility-preserving therapies in EEC.