肿瘤领域文献阅读汇编

Oligometastatic and oligoprogressive disease treated with stereotactic ablative radiotherapy (SABR) represents a clinically heterogeneous entity. Increasing evidence suggests that anatomical definitions alone may not adequately capture underlying biological diversity. This systematic review aimed to synthesize translational evidence exploring evolutionary dynamics, resistance mechanisms, and biomarker-driven stratification in patients treated with SABR. A systematic literature review was performed including prospective and retrospective studies evaluating translational biomarkers in oligometastatic or oligoprogressive settings treated with SABR. Studies assessing genomic, transcriptomic, circulating or immune-related biomarkers were included. Data were summarized qualitatively according to predefined translational domains: (i) evolutionary dynamics under systemic therapy pressure, (ii) baseline biological stratification, (iii) longitudinal circulating biomarkers, and (iv) systemic immune remodeling. Exploratory quantitative visual syntheses were performed using reported hazard ratios when conceptually comparable endpoints were available. 19 studies comprising 1,527 patients were included. Across tumor types and treatment contexts, translational analyses consistently indicated that anatomically defined oligometastatic states encompass biologically distinct subgroups with different risks of systemic progression. Studies evaluating oligoprogression under ongoing systemic therapy suggested a distinction between spatially constrained resistance and systemic molecular escape, supported by circulating tumor DNA and tissue- or plasma-based molecular profiling (including genomic and transcriptomic analyses). Baseline biological features, including adverse genomic signatures and circulating biomarkers, were associated with inferior progression outcomes despite metastasis-directed therapy. Longitudinal biomarkers provided early signals of treatment response and systemic control. Immune remodeling after SABR showed context-dependent effects, both systemic immune activation and treatment-related immunosuppression reported across studies.

Statins and ezetimibe are widely used and safe lipid-lowering therapies for the prevention of atherosclerotic cardiovascular disease. However, concerns persist, especially among the lay public, about the possibility of a long-term association with cancer incidence and cancer-related mortality. To evaluate the association between statin therapy, statin intensity, ezetimibe, and statin-ezetimibe combination therapy with cancer incidence and cancer-related mortality using a network meta-analysis of randomized controlled trials. We conducted a systematic review and Bayesian network meta-analysis of randomized controlled trials comparing statin monotherapy, statin-ezetimibe combination, or ezetimibe monotherapy with no lipid-lowering therapy (no-LLT), including head-to-head comparisons of statin intensities. PubMed/MEDLINE, Embase, and Cochrane were searched through February 2026. Treatment effects were estimated using random-effects Bayesian models and expressed as risk ratios (RRs) with 95% credible intervals (CrIs). A total of 69 randomized controlled trials, including 238,533 participants, were analyzed, of whom 199,286 received statins, 35,836 received statins plus ezetimibe, and 3,411 received ezetimibe alone. For cancer incidence, high-intensity statins (RR 0.95, 95% CrI 0.87-1.03), moderate-intensity statins (RR 1.01, 95% CrI 0.96-1.05), low-intensity statins (RR 1.06, 95% CrI 0.72-1.61), and moderate-intensity statin plus ezetimibe (RR 1.04, 95% CrI 0.96-1.14) were not associated with a reduction compared with no-LLT. For cancer mortality, no significant differences were observed with high-intensity statins (RR 0.94, 95% CrI 0.79-1.09), moderate-intensity statins (RR 1.00, 95% CrI 0.92-1.08), low-intensity statins (RR 1.70, 95% CrI 0.50-6.06), moderate-intensity statin plus ezetimibe (RR 1.12, 95% CrI 0.97-1.32), or ezetimibe alone (RR 1.32, 95% CrI 0.82-2.15), all compared with no-LLT. All head-to-head comparisons among active treatment nodes (statin intensities, statin-ezetimibe combination, and ezetimibe monotherapy) were similarly null, with point estimates close to unity and 95% CrIs crossing 1.0 for all comparisons. Node-splitting revealed no significant inconsistency between direct and indirect evidence for any evaluable comparison. Subtype analyses for lung, breast, gastrointestinal, genitourinary, hematological, skin, and central nervous system cancers showed no significant differences across treatment strategies. In this network meta-analysis of randomized trials, statin therapy across all intensities and ezetimibe, alone or in combination, were not associated with an increased risk of cancer incidence or cancer-related mortality, supporting their oncologic safety within the duration of follow-up available in randomized clinical trials.

This systematic review and meta-analysis included 34 studies encompassing 983 438 adult cancer survivors and evaluated the performance of both general-population and cancer-specific cardiovascular risk-prediction models across vascular events, cancer therapy-related cardiac dysfunction or heart failure, arrhythmias, and composite cardiovascular event outcomes. In total, 27 unique risk scores were assessed. Approximately one-third of model-outcome evaluations demonstrated acceptable-to-good discrimination, defined as an area under the curve of ≥0.70. Among general-population models, four risk scores demonstrated acceptable-to-good discrimination for vascular events, three models were acceptable for CTRCD/heart failure, and two models for arrhythmias. The New Zealand CVD score was the only composite outcome model with an AUC ≥0.70. Cancer-specific scores, such as HFA-ICOS and the Ezaz score, showed high specificity for selected outcomes and may support rule-in decisions, although their lower sensitivity limits use for ruling out risk of CTRCD/heart failure. In our subgroup analysis, model performance varied by cancer type, with more consistent discrimination in breast cancer cohorts and substantially poorer performance in survivors of haematologic malignancies, indicating limited transportability across cancer populations. In summary, these findings provide a comprehensive overview of available cardiovascular risk-prediction tools and may assist clinicians in selecting appropriate models for primary cardiovascular prevention in cancer survivors while highlighting the need for exposure-aware, externally validated models tailored to specific cancer populations.

Malignant ascites is a devastating complication of several advanced malignancies. Beyond serving as a poor prognostic indicator, the rapid accumulation of ascitic fluid imposes a significant symptom burden that can severely worsen quality of life. Although immunotherapy has revolutionized solid tumor oncology, these approaches have shown only limited benefit in the treatment of malignant ascites. Here, we summarize landmark clinical trials exploring immunotherapy as a potential treatment of malignant ascites. We then conceptualize malignant ascites as a functionally distinct immunologic compartment rather than a passive fluid reservoir, highlighting unique immunologic features that may underlie the challenges in translating immunotherapy to this context. By advancing our collective understanding of this unique immune microenvironment, it may become possible to develop rational combination strategies that provide symptomatic relief and meaningfully improve quality of life for patients with end-stage disease.

Cancer management remains fragmented across its continuum, from late-stage diagnosis and salvage therapies to non-personalized surveillance. Here, we present Oncoformer, a unified multimodal transformer model trained on the China Oncology Multimodal Prediction and Surveillance Study (COMPASS) cohort (3.67 million individuals, 17.7 million clinical visits) and validated on independent external cohorts, including the UK Biobank. Oncoformer integrates longitudinal electronic health records with chest X-ray imaging to address multiple clinical tasks: pan-cancer diagnosis (area under the receiver operating characteristic curve [AUROC] = 0.956), future cancer prediction up to 1 year before diagnosis (AUROC = 0.869), tumor stage inference (mean AUROC > 0.90), patient-specific treatment-response forecasting, and recurrence-free survival stratification across ten cancer types (all p < 0.01). Staging predictions were independently validated against postoperative pathological endpoints and shown to converge on core cancer genomic pathways. By translating routine clinical data into a dynamic view of cancer evolution, Oncoformer provides a framework for risk-informed cancer prediction and treatment stratification using routine clinical data.

Immune-related cutaneous adverse events (ircAEs) complicate management in many cancer patients receiving checkpoint inhibitor (CPI) immunotherapy. A new approach to early prognosis and ircAE treatment monitoring is needed in order to improve patient adherence to CPI immunotherapy. Skin tape strips (STS) and plasma were collected prospectively from 22 oncology patients prior to starting CPI and after 6 weeks of CPI therapy. Patients were then followed for ircAE development for 6 months. Ten patients subsequently developed ircAEs. STS were analyzed for cytokines and lipids, while plasma was analyzed for cytokines. A significant increase in stratum corneum (SC) ceramides with short chain fatty acids was observed in patients who developed ircAEs (p < 0.05 compared to non-ircAE groups). This lipid abnormality was already present in the skin of future ircAE patients before starting CPI immunotherapy, and was further dysregulated on CPI therapy. Following CPI initiation, significant increases in skin (but not plasma) IL-8, IL-18, and IP-10 (CXCL10), IL-12, IFNa, MDC (CCL22), and TARC (CCL17) production were observed in patients who subsequently developed ircAEs (p < 0.05 compared to non-ircAE groups). STS analysis of patients developing ircAEs demonstrated SC lipid abnormalities prior to CPI treatment. When studied after 6 weeks of CPI therapy, these SC barrier lipid abnormalities were worsened. In addition, significant increases in proinflammatory cytokine levels were identified in the skin, but not in plasma. These findings point to mechanisms of ircAE development and may allow early identification of patients at risk of developing ircAEs for close monitoring and future prevention of treatment-limiting ircAEs.

Anti-Müllerian hormone (AMH) is widely used to assess gonadotoxicity related to anti-cancer treatments and to measure ovarian function in cancer survivors. The extent of AMH decline across malignancies and treatment regimens and the degree of subsequent recovery have not been systematically quantified. The primary objective of this review was to quantify the change in AMH following cancer treatment and to model longitudinal AMH trajectories in survivors. The secondary objective was to compare AMH trajectories in cancer survivors with those of age-matched non-cancer controls. Data on long-term ovarian reserve after cancer treatment remain limited. This study integrates recent evidence and age-matched control data to better distinguish treatment-related AMH decline from normal reproductive ageing. A systematic search was performed across CENTRAL, MEDLINE, Google Scholar, Embase, and MedRxiv from inception until 24 January 2024, with an updated search on 19 March 2026. Both MEDLINE and Embase were searched via the Ovid platform, whilst Emtree index terms were used for EMBASE. The search strategy combined both index terms and free-text terms: (gonadotoxicity OR premature menopause OR premature ovarian insufficiency (POI) OR premature ovarian failure (POF)) AND (AMH OR Antimullerian* OR Anti-Mullerian) AND cancer. Studies were included if they involved female cancer patients of any age, including paediatric, adolescent and young adult (AYA), and adult populations, who received chemotherapy and who reported either pre- and post-treatment AMH levels or AMH measurements at defined time points after treatment. Studies were excluded if they did not report quantitative AMH data, involved male patients or non-human subjects, or included studies unrelated to cancer treatment-induced gonadotoxicity. The primary outcome was percentage change in AMH from baseline to 12 and 24 months from the end of treatment. Secondary outcomes included long-term AMH trajectories in adult and paediatric cancer survivors compared to the age-related decline in healthy controls. Risk of bias was assessed using the Joanna Briggs Institute (JBI) study-design-specific critical appraisal tools, and certainty of evidence was evaluated using GRADE. Analyses were performed in GraphPad Prism v10.6. Across all four cohorts (breast, lymphoma, thyroid, and paediatric/AYA mixed cancers), AMH levels were higher at 12 and 24 months compared with 6 months from the end of treatment. With breast cancer (median age 37.8 years, n = 2259), AMH was reduced by 76.4% at 12 months (95% CI: -93.5 to -55.7; 24 studies; GRADE: moderate-certainty evidence). With lymphoma, ABVD treatment regimens (median age 24 years, n = 146) were associated with a smaller reduction in AMH at 12 months (-21.1%; 95% CI: -81.6 to +14.8; four studies; GRADE: low-certainty evidence), whereas non-ABVD regimens (median age 26.3 years, n = 188) were associated with a larger reduction (-90.1%; 95% CI: -96.1 to -68.3; six studies; GRADE: moderate-certainty evidence). With thyroid cancer (median age 32.5 years, n = 181), radioactive iodine treatment was associated with a 57.9% reduction in AMH at 12 months (95% CI: -65.2 to -36.4; 4 studies; GRADE: high-certainty evidence). In the paediatric and AYA mixed cancers cohort (median age 9, n = 176), AMH was reduced by 63.1% at 12 months (95% CI: 64.3-61.9; four studies; GRADE: very low-certainty evidence). Among the adult and paediatric survivors who retained ovarian function, AMH trajectories remained relatively stable and closely aligned with age-matched healthy controls. AMH levels consistently improved between 6 and 12-24 months from the end of cancer treatment. We provide generalizable estimates of AMH trajectories in female survivors of both childhood and young adult cancer, indicating that their long-term AMH declines are similar to those of age-matched controls. PROSPERO registration number: CRD420251000939.

The gut microbiota is a critical determinant of both therapeutic efficacy and immune-related toxicity during cancer immunotherapy with immune checkpoint inhibitors (ICIs). Fecal microbiota transplantation (FMT) has emerged as a strategy to introduce beneficial microbial functions, yet clinical outcomes remain variable. In this Review, we integrate evidence from recent clinical trials combining FMT with ICIs in a mechanism-based framework for understanding this variability and to guide safer clinical applications. We discuss how specific microbial functional programs can either buffer or lower thresholds for immune dysregulation and outline implications for donor selection, longitudinal monitoring, and trial design in oncology.

Timothy Yap, MBBS, PhD, vice president and head of clinical development in the Therapeutics Discovery Division and professor in the Department of Investigational Cancer Therapeutics (Phase I Program) at The University of Texas MD Anderson Cancer Center in Houston, spoke with Cancer Discovery's Suzanne Rose about how clinical trials have evolved, the role of AI, and the challenges facing today's drug developers. His research focuses on the first-in-human and combinatorial development of molecularly targeted agents, immunotherapies, antibody drug conjugates, and radioconjugates.

Children with cancer develop many short- and long-term side-effects of treatment1, but the amount of DNA damage associated with chemotherapy exposure is unclear2. Here we used mutational signatures to measure this damage using whole-genome-sequenced tumours from a multi-institutional cohort for which therapy dose and total exposure were uniformly collected3-5. Chemotherapy and radiotherapy were the only exogenous mutagens in relapsed childhood tumours and were often the dominant source of DNA alteration. Compared with treatment-naive tumours, post-therapy cancers carried nearly three times the number of private signatures, and two times the total burden of somatic mutations. Further, the mutagenic effects of different chemotherapies varied. Platinum-based therapies, for which we more than doubled the number of associated signatures, led to the highest number of variants in most patients. Using therapy exposure dates to track when therapy-associated mutations become detectable, we defined a minimum threshold for platinum-associated mutations to emerge. Remarkably, more than one-third of tumours treated with platinum drugs displayed detectable platinum signatures within one year. This work provides genomic evidence for the critical mutagenic effects of chemotherapy in childhood cancer, as a specific driver of tumour evolution. These data highlight opportunities for treatment de-escalation and the future possibility of tracking resistant clones before expansion.

The MYC oncogene family constitutes a master regulatory hub in tumorigenesis, with functional complexity extending far beyond individual gene activities. Recent advances unveil cooperative yet context-dependent antagonism and dynamic interplay among MYC family members, fundamentally reshaping our understanding of lineage specific oncogenic programs. This review synthesizes emerging insights into MYC orchestrated tumor microenvironment remodeling, reciprocal regulation with noncoding RNAs, and the transcriptional and epigenetic governance of metabolic reprogramming. We delineate mechanisms by which MYC drives therapeutic resistance and critically evaluate current strategies targeting MYC or its downstream networks, encompassing direct MYC-MAX disruptors, upstream pathway inhibitors, synthetic lethality, and combinatorial regimens with immune checkpoint blockade or conventional chemotherapy. We further discuss MYC's prognostic significance across diverse cancer types, the critical gap between preclinical efficacy and clinical outcomes, and emerging combination strategies aimed at overcoming acquired drug resistance. By integrating these rapidly evolving biological dimensions, we posit MYC as a highly multidimensional regulatory node whose context-dependent functions present both formidable challenges and promising new opportunities for effective therapeutic intervention.

Macrophages are sentinel innate immune cells that arise from embryonic precursors and bone marrow monocytes, displaying a functional continuum that transcends the classical M1 (pro-inflammatory)/M2 (anti-inflammatory) dichotomy. Under homeostatic conditions, balanced M1/M2 polarization preserves tissue integrity by coordinating immune surveillance, efferocytosis, and tissue repair. When this equilibrium is disrupted, however, M1-skewed responses drive chronic inflammation and autoimmunity, whereas M2-skewed polarization facilitates tumor immune evasion and organ fibrosis. Although diverse therapeutic strategies-including reprogramming, depletion, blockade of monocyte recruitment, CAR-M cells, and nanomedicine-are being explored to restore homeostasis, clinical translation remains constrained by the lack of pathogenic subset-specific markers, insufficient predictive biomarkers for patient stratification, species divergence between mice and humans, and the temporal complexity of context-dependent intervention windows. In this Review, we systematically delineate macrophage plasticity and the molecular mechanisms underlying polarization imbalance, evaluate existing and emerging macrophage-directed interventions, and dissect these translational bottlenecks in depth, highlighting how single-cell multi-omics, humanized models, and dynamic biomarkers can overcome them. By providing a roadmap for precisely calibrating macrophage functional states and restoring M1/M2 balance, this framework will accelerate the development of precision immunotherapies aimed at re-establishing immune homeostasis across a broad spectrum of human pathologies.

Glucose Transporter 1 (GLUT1) is the quintessential facilitator of basal glucose uptake, indispensable for maintaining cellular energy homeostasis, particularly across the blood-brain barrier. Beyond physiological necessity, GLUT1 dysregulation drives a broad pathological spectrum. While genetic haploinsufficiency precipitates severe neurological energy crises like Glut1 deficiency syndrome (Glut1DS), oncogenic networks hyperactivate GLUT1 as the central executor of the Warburg effect to fuel malignant proliferation. Despite its immense therapeutic potential, severe on-target toxicity in normal tissues and adaptive metabolic plasticity remain critical roadblocks to systemic GLUT1 inhibition. This review comprehensively synthesizes GLUT1's multidimensional regulatory networks in health and disease, dissecting how its overexpression fundamentally remodels the tumor microenvironment (TME). We elucidate how GLUT1-driven "metabolic competition" fosters metabolic immune exclusion and drives therapeutic resistance. Furthermore, we map the paradigm shift from traditional systemic blockades to emerging precision interventions. Specifically, we highlight "Trojan horse" glycan-functionalized nanocarriers, targeted protein degradation technologies like PROTACs, and metabolically engineered CAR-T cells. By conceptualizing GLUT1 as the linchpin of the immunosuppressive ecosystem, this work provides a strategic roadmap for precision metabolic immuno-oncology, guiding the development of novel therapies that maximize durable efficacy while minimizing collateral physiological damage.

Macrophages orchestrate immune responses through remarkable phenotypic plasticity, which is intrinsically linked to their ability to reprogram intracellular metabolic pathways in response to microenvironmental cues. While recent advances have highlighted the role of aberrant macrophage metabolism in diverse diseases, a systematic synthesis integrating both intracellular and extracellular metabolic signals remains lacking. This review provides a comprehensive framework for understanding how core metabolic pathways-glycolysis, the TCA cycle, oxidative phosphorylation (OXPHOS), fatty acid oxidation (FAO), and amino acid metabolism-are rewired during macrophage polarization under the orchestration of upstream signaling cascades, including NF-κB, PI3K/AKT/mTOR, JAK-STAT, and MAPK. We examine how exogenous metabolites such as succinate, itaconate, lactate, and amino acids reciprocally regulate macrophage function and discuss tissue-specific metabolic signatures of macrophage subsets-including alveolar macrophages （AMs), Kupffer cells (KCs), and tumor-associated macrophages (TAMs)-in the context of obesity, Type 2 diabetes (T2D), metabolic dysfunction-associated steatotic liver disease (MASLD), infections, autoimmune disorders, and cancer. We further evaluate emerging therapeutic strategies targeting macrophage metabolism, summarizing preclinical and clinical advances across signaling pathways, metabolic nodes, cytokines, and cell-based therapies with detailed trial data. By integrating cell-intrinsic metabolic circuitry with extracellular signals, this review establishes a theoretical foundation for metabolism-targeted immunotherapies and identifies key knowledge gaps for future investigation.

Metastasis remains the leading cause of cancer-related mortality and is increasingly recognized as a consequence of dynamic interactions between tumor cell plasticity and a heterogeneous tumor microenvironment (TME). Rather than being genetically fixed, cancer cells exhibit phenotypic flexibility, enabling reversible transitions among epithelial, mesenchymal, and stem-like states in response to intrinsic programs and extrinsic microenvironmental cues, central to this adaptability. This review synthesizes emerging evidence that tumor progression is governed by reciprocal feedback loops between plastic tumor cells and distinct microenvironmental niches, including hypoxic cores, invasive margins, and perivascular regions. We highlight how stromal components, immune infiltrates, endothelial cells, and extracellular matrix (ECM) remodeling dynamically shape tumor cell states through biochemical and biophysical signals. Advances in single-cell and spatial transcriptomic technologies have revealed the spatial organization and reversibility of these plastic phenotypes, uncovering rare but clinically significant drug-tolerant persister populations. Importantly, we discuss plasticity-mediated therapy resistance as an adaptive, nongenetic process driven by transcriptional and epigenetic reprogramming, metabolic flexibility, ECM stiffening-induced mechanotransduction, and immune-checkpoint plasticity under therapeutic pressure. Together, these findings establish tumor plasticity and microenvironmental heterogeneity as an integrated, evolving system that fuels metastasis and limits durable treatment responses. Targeting this tumor-TME plasticity axis represents a promising strategy to disrupt metastatic progression and overcome therapeutic resistance.

Sirtuins (SIRT1-SIRT7) are nicotinamide adenine dinucleotide (NAD+) dependent deacylases that serves as metabolic sensors, coupling cellular energy status to chromatin structure, mitochondrial function, and stress responses. Dysregulated SIRT activity has been extensively studied in aging, metabolic syndrome, cardiovascular disease, neurodegeneration, cancer, and immune disorders. However, robust human evidence and SIRT-targeted therapies are lacking. Transgenic mouse models serve as key platforms to study gene function and guide therapeutic development. This review synthesizes evidence from Sirt1-7 transgenic mouse models regarding the core cellular processes governed by SIRTs: metabolism, genome integrity, stress resistance, immunity, and autophagy, and illustrates their operation across different organ systems. By comparing global, tissue-specific, and inducible knockout (KO) and overexpression (OE) models of cardiovascular, respiratory, digestive, nervous, endocrine, urogenital, musculoskeletal, malignant, and immune diseases, we identified central regulatory SIRTs (SIRT1, SIRT3, and SIRT6), context-dependent modifiers (SIRT2, SIRT4, SIRT5, and SIRT7), and their organ- and cell type-specific functions. We also summarize representative small-molecule SIRT activators, inhibitors, and degraders, covering both clinical and preclinical studies, and highlight where contradictions and knowledge gaps remain. Together, these analyses help clarify which aspects of SIRT modulation are most promising and under what isoform, tissue, and disease contexts they should be pursued for the development of SIRT‑targeted therapies in human disease.

Mitochondria are essential for cellular homeostasis, integrating various signals to control key cellular functions such as metabolism, apoptosis, inflammation, cell proliferation and redox balance. Given their multifaceted functions, it is not surprising that mitochondrial dysfunction has been implicated as a key contributor to the pathogenesis of numerous human diseases. Consequently, preserving mitochondrial integrity and functionality is vital for overall organismal health. Mitochondrial health is safeguarded by a sophisticated and tightly regulated network of quality control systems. These include mitochondrial proteostasis, which ensures proper protein folding and degradation; mitochondrial biogenesis, which governs the synthesis of new mitochondria; mitochondrial dynamics, encompassing fusion and fission processes; and mitophagy, the selective autophagic removal of damaged mitochondria. Additionally, these core systems are intricately connected to other crucial mitochondrial processes, such as the maintenance of mitochondrial DNA integrity, the regulation of cristae architecture, and the control of mitochondrial permeability transition, all of which are indispensable for optimal mitochondrial performance. Preclinical and clinical studies consistently demonstrate a strong link between impairments in these quality control mechanisms and both aging and the development of a wide spectrum of diseases. These include cancer, metabolic disorders, cardiovascular conditions, neurodegenerative diseases and autoimmune pathologies. In this review, we explore the different facets of mitochondrial quality control and discuss their implications in disease progression and aging. Furthermore, we highlight recent advances in interventions and therapies aimed at modulating mitochondrial quality control, providing an overview of their potential to mitigate disease burden and promote healthy aging.

Exhausted CD8+ T cells (Tex) within the tumor microenvironment (TME) represents a critical barrier limiting anti-tumor immune responses. Tex cells are characterized by upregulated inhibitory immune checkpoint receptors, reduced cytotoxicity, and functional heterogeneity. Their genomic features and regulatory networks remain poorly defined, and only a minority of patients respond to immune checkpoint blockade (ICB) therapy. Single-cell RNA sequencing (scRNA-seq), through high-resolution transcriptomic profiling, has revealed diverse Tex subpopulations, identified subpopulation-specific marker genes and regulatory pathways. Spatial transcriptomics has further mapped the spatial distribution of Tex and their interaction networks with immune cells, tumor cells, and stromal cells, elucidating the impact of spatial heterogeneity on Tex functionality. Current studies indicate that the exhausted state of Tex is dynamic and modifiable, with functional differences among subpopulations closely associated with tumor progression and therapeutic response. However, the genomic characteristics, epigenetic regulation, and spatial interaction mechanisms of Tex require further exploration. This review summarizes recent advances in high-resolution omics technologies for precisely dissecting Tex heterogeneity, functional features, and interactions with other cells. It emphasizes the central value of optimizing Tex-targeted tumor immunotherapy strategies, providing theoretical foundations and directional guidance for developing more effective anti-tumor immunotherapies.

Aging is characterized by progressive physiological decline and accumulation of senescent cells that drive chronic "inflammaging" through the senescence-associated secretory phenotype (SASP). The complement system, traditionally viewed as a systemic extracellular defense mechanism, is now recognized as an essential intracellular network (the complosome). This review synthesizes current research on how intracellular C3 (intC3), and intracellular C5 (intC5) in certain contexts engage in extensive crosstalk with the mTOR, NF-κB, and AMPK pathways to modulate core cellular processes. We detail how intC3 intersects with multiple canonical hallmarks of aging-including cellular senescence, mitochondrial dysfunction, proteostasis loss, genomic instability, epigenetic alterations, altered intercellular communication, stem cell exhaustion, and deregulated nutrient sensing. Furthermore, we examine organ-specific consequences of intC3 dysregulation across the aging brain, liver, eye, kidney, vasculature, lung, and immune system, as well as cancer. Finally, we discuss therapeutic strategies-including complement inhibitors, senolytics, senomorphics, RNA interference, proteolysis targeting chimeras (PROTACs), and nanotechnology-enabled delivery-and outline critical gaps in compartment-specific tools, conditional knockouts, and longitudinal models. Precision strategies that preserve systemic complement function while selectively modulating intracellular pools will be essential to translate complosome biology into durable interventions for age-related diseases.

Metastatic microenvironments vary widely not only in their biochemical composition but also in their mechanical properties. Here, we examined how the mechanical rigidity of the metastatic niche affects metastases seeding and the local efficacy of antitumor immunosurveillance. Cancer cells stiffened in response to increasing environmental rigidity, a biophysical change that mechanically sensitized them to killing by cytotoxic lymphocytes. In immunodeficient mice, rigidity sensing by cancer cells yielded robust bone colonization, accompanied by marked stiffening of the cancer cells themselves. Conversely, in immunocompetent hosts, stiffer cancer cells were selectively eliminated, and bone metastasis was suppressed. In patients, metastatic cell stiffness was associated directly with environmental rigidity and inversely with immune infiltration. Expression of Spp1, encoding the secreted glycoprotein osteopontin, defined a subset of cancer cells that expanded in the bone, and deletion of Spp1 limited environmentally induced cancer cell stiffening, bone colonization, and immune vulnerability. Thus, environmental mechanosensing regulates both metastases seeding and antitumor immunity, providing an immunological basis for metastatic site selection.

The development of simple, efficient, and actively targeted drug delivery systems remains a significant challenge for clinical translation. Here, we propose a carrier-free nanodrug platform based on polyphenol-modified prodrugs with self-assembly and ligand-adsorption capabilities. Specifically, two gallic acid-modified Cabazitaxel (CTX) prodrugs were synthesized: a reduction-responsive prodrug (CSG) and a non-responsive control (CCG). These prodrugs spontaneously assembled into nanocomplexes, and transferrin (Tf) was non-covalently adsorbed onto their surfaces via polyphenol-protein interactions, forming Tf-functionalized nanocomplexes (Tf NCs). This ligand-anchoring strategy endowed the system with Tf receptor (TfR)-mediated active targeting and enhanced colloidal stability. Notably, the CSG@Tf NCs exhibited a cascade targeting effect: prolonged circulation, tumor-specific accumulation via TfR recognition, and glutathione (GSH)-triggered CTX release in the tumor microenvironment. In vitro and in vivo studies confirmed that CSG@Tf NCs significantly improved antitumor efficacy while reducing systemic toxicity. This work presents a novel, carrier-free, and dual-functional strategy for constructing targeted prodrug nanomedicines with strong translational potential.

Organic near-infrared II (NIR-II) phototheranostic agents hold great promise for tumor imaging and phototherapy. However, their development is fundamentally challenged by the coupled and competing optical pathways in conventional designs: rigid planar fluorophores that excel in the molecular state often suffer from severe aggregation-caused quenching (ACQ) in nanoparticles, leading to drastic losses in both fluorescence and reactive oxygen species (ROS) generation. To decouple these optical functions, we report a modular copolymer-based anti-quenching strategy by integrating a flexible segment with a hydrogen-bond-enforced rigid segment. The key to decoupling lies in the tunable segment ratio, which allows composition-dependent modulation with reduced trade-offs of NIR-II fluorescence, ROS production, and photothermal conversion. As the rigid-segment fraction increases in the copolymer series (F8R2, F5R5, and F2R8), NIR-II fluorescence and ROS generation are markedly enhanced, while the photothermal conversion efficiency is only slightly compromised. Remarkably, the F2R8 nanoparticles (80% rigid segment) retain >60% of their NIR-II fluorescence and high ROS productivity in the aggregated state, starkly contrasting the >90% quenching observed in conventional ACQ-type small molecules. This effective partial decoupling of optical functions and suppression of ACQ enable high-contrast NIR-II fluorescence imaging-guided combined phototherapy in vivo. Our work provides a general design paradigm based on copolymer modularity for developing high-performance quenching-resistant organic phototheranostics.

Messenger RNA (mRNA) technology has emerged as a cornerstone in vaccine development and therapeutic applications, offering key benefits such as high potency, rapid scalability, and cost-effectiveness. The success of COVID-19 mRNA vaccines has underscored their efficacy and safety. However, residual byproducts generated during mRNA synthesis, such as unincorporated caps, nucleoside triphosphates (NTPs), DNA templates, enzymes, abortive transcripts, and double-stranded RNA (dsRNA), pose significant challenges to the clinical application of the RNA therapy. Among these, dsRNA is particularly problematic as it can activate various innate immune responses, suppress mRNA translation and potentially compromise the therapeutic efficacy of mRNA. Therefore, effectively removing dsRNA from in vitro synthesized mRNA is essential before its used in preclinical or clinical settings. In this review article, we provide a comprehensive overview of current mRNA development pipelines and ongoing clinical trials, and recent advances in mRNA purification techniques. Specifically, we focus on strategies for dsRNA removal, which can be broadly categorized into two approaches: (1) separating or removing dsRNA from in vitro transcription (IVT) mRNA products using methods such as RP-HPLC chromatography and cellulose-based purification; and (2) minimizing dsRNA formation during IVT by employing engineered RNA polymerase mutants, chaotropic agents, and magnetic beads, as well as modifying/optimizing DNA templates or RNA molecules to reduce dsRNA generation. We also discuss the advantages and limitations of these purification methods, the factors influencing the selection of purification strategies, and explore potential future directions for improving dsRNA purification technologies and their applications in mRNA-based therapeutics.

Immunotherapy is now an established and efficient treatment option for many cancer patients. However, a proportion of patients still experience poor outcomes due to treatment resistance. Thus, a clear understanding of key mechanisms of resistance is needed for the development of new treatments. Here, we employ mouse models to explore an immunotherapeutic approach based on anti-MARCO (αMARCO) and anti-PD-L1 (αPD-L1) antibody-mediated targeting of tumor-associated macrophages (TAM). We demonstrate that effective immunotherapy relies on a functional endosomal pattern recognition machinery. We determine that endosomal Toll-like receptors (TLR), specifically TLR9, precondition macrophages to respond to αMARCO treatment by regulating the transcription of inflammasome components. Absence of TLRs renders TAMs unresponsive to treatment while retaining an immunosuppressive phenotype. Thus, we uncover the intracellular TLR signalling as a feature of immunotherapy efficacy, required to sensitise TAMs to treatment, and indicate that TLR targeting could be exploited to improve immunotherapeutic outcomes.

Natural killer (NK) cells act as the host's first line of immunosurveillance, and their maturation and effector functions are finely regulated by multiple factors. Although histone deacetylase (HDAC) inhibition has been reported to impair NK cell viability and/or antitumor effects, the specific HDAC member involved and the underlying mechanisms remain unclear. We analyzed the expression of HDAC family members during NK cell activation and in tumor-infiltrating NK cells. Genetic approaches, including Hdac6 knockout, knockdown, and overexpression, were used to evaluate its role in NK cell proliferation, maturation, and effector function. Mechanistic studies examined protein acetylation status, chaperone activity, and IL-15-STAT5 signaling. Functional relevance was assessed using NK cell adoptive transfer models and interleukin (IL)-15 combination treatment in tumor settings. HDAC6 was the most significantly upregulated HDAC member on NK cell activation, whereas its expression was reduced in tumor-infiltrating NK cells and correlated with poor patient survival. Hdac6 deficiency markedly impaired NK cell proliferation, maturation, and effector functions, while Hdac6 overexpression showed a promoting effect. Mechanistically, loss of Hdac6 led to the hyperacetylation of HSP90, which in turn destroyed its chaperone activity for phosphorylated STAT5 and blocked the transduction of the IL-15 signaling pathway indispensable for NK cell maturation and effector function. Translationally, HDAC6 overexpression not only obviously improved tumor suppression of transferred NK cells, but also synergized with IL-15 treatment to further boost NK cell antitumor efficacy. HDAC6 was identified as a critical regulator of NK cell biology through strengthening the IL-15/STAT5 pathway and may serve as a potential target for improving NK cell function.

Conventional cancer therapy has largely been organized around maximal cytotoxic elimination, yet intense tumor killing can impose evolutionary selection pressures that promote resistance, clonal escape, and relapse. Here, we propose therapeutic super-competitor cells (TSCs) as a programmable, evolution-aware extension of educative cancer care rather than as an established clinical platform. In this framework, TSCs are engineered cells designed to home to or be locally positioned within tumor-associated niches, sense malignant ecological cues, transiently acquire bounded competitive fitness, restrict malignant access to space, metabolites, and stromal support, and then undergo monitored clearance or benign differentiation. TSCs are therefore not defined by direct target-cell cytolysis, although competitive pressure, metabolic restriction, or immune remodeling may secondarily reduce tumor viability. This Review integrates concepts from cancer ecology, tumor microenvironment biology, developmental cell competition, immunotherapy, and synthetic biology while distinguishing established principles from speculative engineering applications, evaluating feasible early disease contexts, and outlining measurable endpoints for ecological replacement, including spatial occupancy, clonal displacement, resource gradients, persistence kinetics, and responsiveness to safety triggers. We further discuss organoid and tumor-on-chip competition assays, spatial and lineage tracking, response biomarkers, therapeutic-window definition, layered containment, and ethical/regulatory safeguards. By reframing engineered cell therapy as a controllable ecological intervention rather than simply a vehicle for maximal killing, the TSC framework provides a testable but still hypothetical direction for educative cancer care and evolution-aware oncology.

Circulating tumour DNA (ctDNA) has emerged as one of the most promising biomarkers in oncology, with potential applications from detection of minimal residual disease (MRD) and diagnosis to prognosis, treatment guidance, and response monitoring. This personal view critically examines recurring methodological and statistical pitfalls in ctDNA studies with longitudinal measurements. We highlight important issues with lead time definition and demonstrate that performance metrics like sensitivity and specificity are inherently time-dependent. Furthermore, we show why immortal time bias arises when classifying patients incorrectly as always positive, based on becoming positive during surveillance in MRD related studies. With a combination of toy data examples, simulations, and re-analysis of published data we provide practice-oriented guidance to support the design, analysis, and reporting of ctDNA trials. This viewpoint paper aims to sensitise the community and raise awareness of pitfalls in longitudinal ctDNA analysis, rather than offer definitive solutions-more evidence is needed first.

Developing radiosensitizing agents to amplify tumor-eradicating effects on primary, regional recurrence, and distant metastases plays a transformative role in modern cancer care. Here, we report the de novo design of biocatalytic artificial metalloenzymes with an IrMn-cluster-based redox center (IMM) to achieve radiosensitized systemic antitumor responses for preventing malignant tumor metastasis and recurrence. Notably, our findings indicate that Mn-organic ligands substantially enrich the electron density of Ir clusters, thereby optimizing their interaction with oxygen species and markedly enhancing the production of both reactive oxygen species and molecular oxygen. When combined with radiotherapy, the IMM effectively amplifies DNA damage and induces pronounced apoptosis by alleviating intratumoral hypoxia. This shift reprograms the tumor microenvironment, enhancing radiosensitivity and facilitating the infiltration and activation of intratumoral CD8⁺ T cells and dendritic cells. Moreover, when integrated with anti-PD-1 therapy, this coordinated therapeutic regimen elicits potent systemic immune responses and durable antitumor memory, effectively suppressing tumor recurrence and metastasis while markedly improving therapeutic efficacy and long-term survival. We anticipate that this conceptual design could offer a promising and translationally relevant nanomedicine platform for radiotherapies.

Clinical translation of nanomedicines remains constrained by a persistent gap between static preclinical assays and the dynamic shear environment of the human vasculature. This review argues that fluid shear stress is an underappreciated contributor to clinical attrition, acting alongside well-established factors including biomarker-target mismatch, enhanced permeability and retention heterogeneity, preclinical-clinical discordance in tumor biology, and manufacturing variability. Circulating nanocarriers traverse a mechanical environment in which wall shear stress spans from 0.1 dyn/cm2 in hepatic sinusoids to over 1000 dyn/cm2 in stenotic regions. Emerging quantitative studies indicate that these forces can modulate carrier stability, protein-corona composition, and drug release in ways that static in vitro assays cannot resolve, with the cholesterol-rigidified bilayer of Doxil and the shear-activated platforms of Korin and colleagues providing representative positive control and mechanistic evidence. We synthesize recent progress at the interface of computational fluid dynamics and machine learning. Physics-informed neural networks, deep operator networks, and graph neural networks feature prominently in this synthesis and underpin an integrated framework for mechanistically informed pharmacokinetic prediction. Reported benchmarks show surrogate models reconstructing hemodynamic fields with under 5% error relative to high-fidelity CFD while reducing data requirements 5- to 10-fold, and convolutional surrogates trained on approximately 1800 patient-specific simulations predicting time-averaged wall shear stress with 2.5% mean absolute error. These surrogate models integrate hemodynamic descriptors (TAWSS, OSI, and RRT) with distributional particle features (D10, D50, and D90). We outline the evidentiary and methodological path toward biomechanically informed nanomedicine development aligned with emerging regulatory frameworks, while acknowledging that routine regulatory acceptance of computational shear modeling remains a future milestone rather than current practice.

Traditional cancer drug discovery encounters challenges, including lengthy synthesis durations, high costs, and a 90% failure rate in clinical trials, primarily due to inadequate chemical design and drug properties. Artificial intelligence (AI) provides powerful computational tools to overcome these issues by speeding up target identification, predicting properties, and optimizing leads. This review emphasizes the influence of new AI platforms like AlphaFold3, molecular interactions are structurally optimized (MISATO), and ZairaChem on the discovery of oncology drugs. We specifically examine how AI reconciles chemical design with pharmacological feasibility. In addition to evaluating these advancements, we meticulously evaluate methodological challenges, including dataset bias, overfitting, insufficient external validation, and reproducibility issues. Furthermore, the development of complex and targeted modalities, such as antibody-drug conjugates (ADCs), aptamer-drug conjugates (Ap‑DCs), and proteolysis-targeting chimeras (PROTACs), is being explored for cancer treatment using AI. Following a detailed review of regulatory and clinical translation issues, this review presents practical tips for improving model validation, data sharing, and incorporation into medicinal chemistry workflows. By examining successes and persistent limitations, this review article offers a strategic roadmap for leveraging AI to provide clinically translatable cancer therapies with enhanced chemical and pharmacological balance.

Although macrophages are a powerful cell-based platform for cancer immunotherapy, their antitumor functions, such as phagocytosis and inflammatory responses, are limited by the immunosuppressive tumor microenvironment. Here, we show that decorating cancer cell membranes with bacteria-derived pathogen-associated molecular patterns (PAMPs) initiates phagocytosis and inflammatory responses of macrophages toward cancer cells involving various pattern-recognition receptor signaling pathways. Bacteria-derived PAMPs were formulated into membrane-decorating nanoparticles, and these nanoparticles reprogrammed immunosuppressive macrophages into inflammatory phenotypes. Cancer cell membrane-attached PAMP nanoparticles maintained their immunostimulatory responses, stimulating macrophages' antitumor functions. The fraction of phagocytic macrophages significantly increased when coincubated with membrane-decorated cancer cells, along with an increased secretion of inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Transcriptomic gene ontology analysis revealed that the response of macrophages to PAMP-decorated cancer cells resembled their response to bacteria, involving signaling pathways including inflammatory response and innate immune response. In a mouse model, locally injected membrane-decorating PAMP nanoparticles suppressed tumor growth. The therapeutic effect was more pronounced in combination with the chemotherapeutic drug doxorubicin. Median survival days significantly increased in both the PAMP nanoparticle and the PAMP nanoparticle plus doxorubicin combination group with complete remission cases, compared to the doxorubicin group. Our findings provide insights into the use of macrophages as a cancer immunotherapy modality.

Conventional type 1 dendritic cells (cDC1s) play a pivotal role in initiating CD8+ T-cell-mediated antitumor immunity, and their intratumoral abundance strongly correlates with immunotherapy efficacy. Here, we demonstrate that orally administered chenodeoxycholic acid (CDCA), a clinically approved metabolic modulator, accumulates in the tumor interstitial fluid (TIF) of tumor-bearing mice as its metabolite taurochenodeoxycholic acid (TCDCA), thereby potentiating the antitumor functions of cDC1s and CD8+ T cells through a gut microbiota-independent mechanism. Mechanistic studies revealed that CDCA mediates this immunomodulatory effect via TGR5 signaling, which enhances cDC1 functionality and subsequently promotes the proliferation and activation of tumor-specific CD8+ T cells. Clinical correlation analysis of human tumor specimens revealed a positive association between Takeda G protein-coupled receptor 5 (TGR5) expression and HLA-A/B/C/E/F/H levels, with elevated TGR5 expression significantly predicting improved overall patient survival. Moreover, CDCA exhibits synergistic antitumor effects when combined with anti-PD-1 immunotherapy and a poly I:C adjuvant, leading to robust tumor growth suppression and enhanced antitumor-immune responses. Overall, our study elucidates a novel immunoregulatory mechanism through which CDCA modulates tumor progression, highlighting CDCA and its bile acid derivatives as promising therapeutic candidates for cancer immunotherapy, providing a potential strategy to overcome resistance in cold tumors by revitalizing the cDC1-CD8+ T-cell axis.

Clinical translation of nanomedicines is greatly hindered by insufficient understanding of their in vivo process, yet a key challenge lies in quantifying the encapsulated versus free drug forms in tissues and cells. Herein, we present a facile, versatile anti-PEG single-chain variable-fragment antibody (PEG-scFv)-based method enabling quantitative measurement of both forms in various biofluids (e.g., interstitial fluid, cytoplasm). By this method, we map the in vivo process of PEGylated liposomal doxorubicin (sLip/Dox) at unprecedented resolution. In the bloodstream, doxorubicin remains largely encapsulated in liposomes (>99%). In liver as the main organ for drug elimination, less drug was distributed in the interstitium (>80% encapsulated) but more in liver cells (mainly in Kupffer cells) released in a time-dependent manner, accompanying doxorubicin transferred to hepatocytes most in free form by 12 h postinjection. After extravasation into tumors, there was a limited access of sLip/Dox to tumor cells, confining most of the drug in the interstitium mainly being encapsulated (more than 75%), and the internalized fraction underwent a gradual release process in both tumor-associated macrophages and tumor cells. These findings revealed that for sLip/Dox, which primarily underwent drug release intracellularly, cellular internalization rates could be the key factor in determining its in vivo performance. Given widespread PEGylation on developing nanomedicines and the cost-effectiveness of scFv production, PEG-scFv offers a broadly applicable tool for dissecting in vivo processes of nanomedicines to establish dose-effect relationships like small-molecule drugs, further to guide rational nanotherapeutic design.

Cancer vaccines have underperformed clinically, largely because antigen-specific CTL priming and expansion are constrained by insufficient costimulatory activation and increased DC-mediated coinhibition. Here, we present a personalized cancer vaccine platform, STAR, which engineers stimulator of interferon gene (STING)-activating autologous tumor-derived microvesicles (TMVs) to concurrently amplify costimulatory signaling and relieve DC coinhibition during antigen cross-presentation. TMVs provide broad tumor antigen cargo and intrinsic STING activity; loading manganese phosphate nanoparticles (MnPs) potentiates cGAS-STING signaling, drives DC maturation, and enhances antigen cross-presentation. Importantly, concurrent delivery of PD-L1-targeting siRNA (siPD-L1) effectively inhibits DC-associated PD-L1, freeing CD80 to engage CD28 and limiting PD-1/PD-L1 signaling and thereby significantly enhancing antigen-specific CD8+ T cells. In mouse models, STAR vaccination elicited durable prophylactic immunity and achieved substantial therapeutic efficacy against melanoma and colorectal tumors without additional immunotherapies. Notably, patient-specific STAR vaccines derived from surgically resected tumors effectively prevented postoperative recurrence and metastasis in aggressive melanoma and triple-negative breast cancer models, thereby highlighting their translational potential. Overall, the STAR vaccine platform offers a versatile and clinically applicable approach to enhancing personalized cancer immunotherapy across diverse cancer types.

Tumor radioresistance is a leading cause of radiotherapy failure. This study presents a facile strategy for enhancing tumor radiosensitivity using a covalent organic framework (COF) constructed within cells. Upon cellular internalization, acetamide and iodoacetal precursors undergo hydrolysis to form amine and iodoaldehyde monomers that spontaneously polymerize and crystallize within the lysosomes. This intracellularly generated iodine-rich COF (in situ UM-113) possesses an ordered structure that effectively facilitates radiation energy deposition, producing localized reactive oxygen species and amplifying DNA damage. In vitro and in vivo, in situ UM-113 suppresses tumor growth by triggering both ferroptosis and mitotic catastrophe. This strategy bypasses the complex synthesis and purification steps required for conventional nanomedicine. Thus, in situ COF construction is a promising strategy for enhancing cancer radiotherapy, offering a modular platform for the intracellular synthesis of functional bionanomaterials.

T cell exhaustion (TCE), a hallmark of chronic infections and cancer, is characterized by progressive loss of effector function, sustained expression of inhibitory receptors, and stable transcriptional/epigenetic reprogramming. Within the tumor microenvironment (TME), exhausted CD8+ T cells fail to eliminate malignant cells, contributing to immune evasion and resistance to immunotherapy. Although checkpoint blockade has provided clinical benefit, outcomes remain variable, underscoring the need to better understand the temporal and mechanistic basis of exhaustion. Current modeling efforts have yielded valuable insights; however, they often focus on isolated aspects of tumor-immune interactions. Deterministic models such as ordinary, partial, and delay differential equations capture population dynamics, but omit stochastic variation and single-cell heterogeneity. Stochastic and agent-based models address randomness and spatial structure at a greater computational cost. Hybrid and multiscale approaches increasingly integrate these methods, but few explicitly capture the progressive, time-series nature of TCE as revealed by recent epigenetic and transcriptomics studies. This review analyzes various mathematical and computational frameworks including deterministic, stochastic, and hybrid approaches that have been applied to study TCE in viral and cancer contexts. We distinguish between TCE-specific models that directly represent exhaustion dynamics and TCE-relevant frameworks that model tumor-immune interactions, spatial tumor microenvironment features, and pharmacological interventions that could be adapted to optimize future TCE models. By comparing strengths and limitations across frameworks, we identify key gaps including limited integration of temporal resolution, lack of multiscale intracellular regulation, and scarce validation with longitudinal experimental data. We also highlight how TCE-relevant models can support pharmacological and translational questions, including dose optimization, pharmacokinetic /pharmacodynamic (PK/PD) integration, and mechanisms of immunotherapy failure. Future models that adopt hybrid, time-resolved, and multiscale designs linking intracellular regulatory networks, population-level signaling, and spatially heterogeneous TME features, calibrated with time-series omics data, would be invaluable in addressing these gaps. Such frameworks would provide mechanistic insights into exhaustion trajectories, supporting advances in immunotherapy design and clinical outcomes.

With increasing awareness of health-oriented antioxidant diets, plant-derived antioxidants (PDAs) such as polyphenolics, terpenoids, vitamins, alkaloids and saponins, are promising alternatives to synthetic antioxidants (SAs). PDAs boost human immunity and health by scavenging reactive oxygen species, activating endogenous antioxidant defenses, and reducing oxidative cell damage. As a result, they are widely used in preventing and treating oxidative stress-related chronic diseases, including neurodegenerative diseases (NDDs), viral diseases, cardiovascular diseases (CVDs), and diabetes, as well as certain cancers. Moreover, due to their antioxidant, coloring and preservative effects, PDAs are utilized in food industry to extend shelf-life, improve food quality and inhibit pathogenic microorganisms. However, obtaining high-quality PDAs remains challenging because of their low natural abundance, complex impurities, poor stability, and prone to contamination and deterioration during isolation. This review highlights recent advances in the classification, sources, antioxidation, biosynthesis and regulation, as well as extraction and refining of PDAs. Their functional applications are summarized in two aspects: (i) as food additives (e.g. antioxidants, colorants and preservatives) to reduce potential risk of SAs and enhance food value-added; and (ii) as biomedicines (with anti-inflammatory, anti-NDDs, antiviral, anticancer, anti-CVDs and anti-diabetic effects), providing new strategies for chronic-disease prevention through multi-target regulation of oxidative-stress pathways. Integration of synthetic biology, green purification and nano-delivery is envisioned to overcome industrial challenges of PDAs, offering new insights for their precise design and versatile application in the healthcare and food industries.

Extracellular vesicles (EVs) are nanoscale, membrane-bound particles that transport diverse biomolecules-including proteins, nucleic acids, lipids, and metabolites-between cells, thereby orchestrating key processes in cancer progression. Tumor-derived EVs modulate angiogenesis, epithelial-to-mesenchymal transition, extracellular matrix remodeling, fibroblast activation, and immune evasion, shaping the tumor microenvironment, and driving metastasis as well as therapy resistance. With their stability in biofluids and cargo reflective of cellular origin, EVs have emerged as powerful non-invasive biomarkers for early detection, disease monitoring, and prognosis across multiple cancer types. Recent advances in enrichment, characterization, and molecular profiling technologies-ranging from ultracentrifugation and microfluidics to proteomics, RNA sequencing, and surface-enhanced Raman spectroscopy, have greatly expanded the diagnostic potential of EVs. Integration with machine learning further enhances sensitivity, specificity, and tumor classification, while multiomic and multiplex platforms enable high-throughput and clinically relevant applications. This review highlights the multifaceted roles of EVs in cancer biology, catalogs emerging biomarkers, and compares state-of-the-art detection technologies, offering a comprehensive reference for advancing EV-based diagnostics and precision oncology.

Whole Slide Image (WSI) analysis, with its ability to reveal detailed tissue structures in magnified views, plays a crucial role in cancer diagnosis and prognosis. Due to their giga-sized nature, WSIs require substantial storage and computational resources for processing and training predictive models. With the rapid increase in WSIs used in clinics and hospitals, there is a growing need for a continual learning system that can efficiently process and adapt existing models to new tasks without retraining or fine-tuning on previous tasks. Such a system must balance resource efficiency with high performance. In this study, we introduce COSFormer, a Transformer-based continual learning framework tailored for multi-task WSI analysis. COSFormer is designed to learn sequentially from new tasks wile avoiding the need to revisit full historical datasets. We evaluate COSFormer on a sequence of seven WSI datasets covering seven organs and six WSI-related tasks under both class-incremental and task-incremental settings. The results demonstrate COSFormer's superior generalizability and effectiveness compared to existing continual learning frameworks, establishing it as a robust solution for continual WSI analysis in clinical applications. The code is released at https://github.com/QuIIL/COSFormer.

The clinical translation of stimulator of interferon genes (STING)agonist-based cancer immunotherapy is limited by uncontrolled and sustained STING activation, which leads to immune exhaustion and systemic toxicity. In this forum, we critically analyze recent advances, including stimuli-responsive, biomimetic, pulsatile, intracellular-triggered release, and synergistic systems that enable precise spatiotemporal STING activation, as well as associated gaps in clinical translation. Finally, we present futuristic perspectives on next-generation STING-based cancer immunotherapy strategies.

The cGAS-STING pathway serves as a central hub for DNA-triggered innate immune activation in tumors. Nevertheless, the clinical translation of stimulator of interferon genes (STING) agonists remains hindered by challenges such as rapid in vivo degradation, inefficient cytosolic delivery, and the risk of systemic inflammation. Inorganic nanomaterials, leveraging their high specific surface area, tunable size and morphology, and unique surface chemical properties, provide an ideal platform for the precise delivery and spatiotemporally controlled release of STING agonists. Furthermore, these materials can induce the release of double-stranded DNA (dsDNA) to activate STING while also synergistically potentiating STING activation through the induction of immunogenic cell death or the release of specific metal ions. This review highlights that the application of inorganic nanomaterials in STING pathway activation extends beyond simple agonist delivery to encompass precise spatiotemporal control and modulation of signal intensity. We systematically outline design strategies for inorganic platforms that facilitate agonist protection and stimuli-responsive release. Furthermore, we discuss their synergistic integration with therapeutic modalities, including radiosensitization, reactive oxygen species (ROS) induced DNA damage, and immunogenic cell death (e.g., pyroptosis and ferroptosis), ultimately contributing to the establishment of a cGAS-STING amplification immune circuit. By achieving synergistic integration of delivery, activation, and potentiation, inorganic nanomaterials enhance both the efficacy and safety of STING-targeted immunotherapy, realizing the integrated functional advantage of "carrier-adjuvant-inducer" and offering a new paradigm for cancer immunotherapy.

The tumor microenvironment (TME) and its complex, dynamic interactions play a pivotal role in cancer development, progression and therapy response. However, faithful recapitulation of the diverse cellular and structural components of the TME in vitro remains a major challenge in cancer research. Traditional 2D cancer cell cultures fail to preserve TME interactions and tissue organization that critically impacts tumor behavior, while advanced 3D systems, including organoids, 3D-bioprinted structures and microfluidic platforms capture only selected aspects of TME complexity and host physiology. Recent advances in the culture of primary tumor specimens with minimal disruption to tissue architecture has led to a rapidly evolving set of model systems that benefit TME research. These ex vivo cultures (EVCs), as we collectively refer to them here, are established from fresh primary tumor tissue and preserve the native tumor architecture, extracellular matrix composition, immune and stromal compartments and their multilayered crosstalk within a physiologically relevant context. This review outlines the historical evolution of 2D and 3D model systems in oncology, followed by a comprehensive overview of current EVC methodologies. Furthermore, we address their applications in fundamental cancer research, personalized medicine and drug discovery, while highlighting their advances and challenges for the future.

Apoptosis is a highly conserved process that eliminates unwanted or damaged cells in both physiological and pathological conditions. Dysregulation of apoptosis leads to developmental abnormalities and various diseases, such as neurodegeneration and cancer. Drosophila inhibitor of apoptosis 1 (Diap1) plays a crucial role in cell survival by inhibiting caspases and preventing apoptosis. However, under stress conditions, the prodeath proteins Rpr, Hid, and Grim (RHG) induce apoptosis by antagonizing Diap1. Despite being a key component of the apoptotic pathway, the mechanism that controls the stability of Diap1 remains unknown. Here, we find that loss of hdac3 results in the activation of apoptosis, which is completely blocked by expressing Diap1. Although Hdac3 localizes in both the cell cytoplasm and nucleus, only the cytoplasmic Hdac3 is able to suppress apoptosis induced by hdac3 deficiency, RHG overexpression, or x-ray irradiation. This finding indicates that Hdac3 exerts an antiapoptotic role independent of its canonical epigenetic functions. Loss of hdac3 decreases Diap1 protein, which is rescued by introducing cytoplasmic Hdac3. The deacetylase activity is necessary for Hdac3 to suppress apoptosis. Mechanistically, Hdac3 interacts with Diap1 to remove the acetyl group from K315 on Diap1, thereby increasing its stability. Compared with the wild-type Diap1, the acetyl-deficient mutant Diap1-K315R exhibits stronger stability and antiapoptotic activity. Last, RHG proteins compete with Hdac3 for Diap1 interaction, directing Diap1 toward degradation and triggering apoptosis. Together, these findings not only reveal the involvement of Diap1 acetylation modification in apoptosis regulation but also clarify the role of Hdac3 in apoptosis.

The rapid growth of quantum computing is driven by promises of performing complex calculations with unprecedented speed; however, current use cases have been limited by quantum hardware and the difficulty of identifying problems that classical computers cannot easily address. Within these constraints, biological problems including drug discovery, protein folding, and precision medicine present an opportunity to understand how current quantum hardware can make advances. In immunology, accurate prediction of cancer neoantigens remains a major challenge, limited by small, noisy datasets and the inability of classical models to generalize. In approaching the problem, we explore multiple noise mitigation techniques, including Pauli twirling and dynamical decoupling, in conjunction with controlled shot-based sampling to stabilize training on real hardware and in a warm start hybrid approach. With these approaches, we demonstrate the use of Quantum Convolutional Neural Networks (QCNNs) for both MHC binding and immunogenicity prediction, including a quantum hardware experiment involving 46 qubits that achieved a 6% increase in classification accuracy with fewer training samples compared to classical approaches. Building on these models, we introduce Quantum Convolutional HLA Immunogenic Peptide Prediction (Q-CHIPP), a combinatorial framework integrating MHC binding and T-cell recognition. It targets HLA-A*02:01-restricted 9-mer peptides and, more accurately, identifies those peptides known to be immunogenic, improving the prognostic impact of predicted neoantigen load. Together, these represent a large-scale application of QCNNs in biomedical modeling, highlighting both the feasibility and promise of quantum machine learning for data-limited biological systems and establishing a scalable foundation for quantum-enhanced biomedical research.

Tumor cells promote metabolic dysregulation of immune cells by controlling the metabolic landscape of the tumor microenvironment. It is unclear whether tumors restrict specific nutrients to drive rapid growth and immune evasion in addition to the overconsumption of nutrients to support anabolism. We identified that up-regulation of solute carrier family 7 member 1 (SLC7A1) increased arginine utilization and promoted tumor growth, whereas down-regulation of SLC7A2 decreased lysine catabolism to support immune evasion. Repression of lysine catabolism in tumor cells reduced glutaconic acid (GC), a medium-chain acyl-CoA dehydrogenase-dependent lysine catabolite that has immunostimulatory effects on antitumor CD8 T cells. GC modified pyruvate kinase M2 (PKM2) through posttranslational glutaconylation at key lysine residues Lys336 (K336) and K337. This modification reinforced PKM2 dimers, transcriptionally driving metabolic reprogramming and reinvigorating antitumor CD8 T cells. Our study highlights an amino acid trade-off that dynamically optimizes the metabolic preferences of tumors to promote proliferation and immune evasion.

Genome-wide CRISPR screens have systematically identified genes required for cancer cell survival, yet these studies are typically performed under standardized conditions that do not fully recapitulate the physiological stresses encountered within the tumor microenvironment. In a recent issue of Nature Genetics, Cheruiyot and colleagues perform genome-wide loss-of-function screens under inflammatory conditions induced by interferon-β (IFN-β), interferon-γ (IFN-γ), and tumor necrosis factor (TNF), revealing that distinct cytokines impose different genetic requirements for tumor cell survival. The study shows that inflammatory signaling reshapes genetic dependency landscape in a cytokine-specific manner. Mechanistic analyses identify the glycosylphosphatidylinositol (GPI) transamidase complex and FITM2 as representative examples of genes that become selectively required under inflammatory stress by maintaining membrane protein maturation, endoplasmic reticulum homeostasis, and resistance to oxidative stress. These findings broaden our understanding of how inflammatory cytokines influence tumor cell biology beyond transcriptional regulation and immune recognition. More broadly, the study highlights the value of incorporating physiologically relevant conditions into functional genetic screens, suggesting that conventional dependency maps capture only part of the genetic requirements for tumor survival. Applying similar approaches to other microenvironmental stresses-including hypoxia, metabolic competition, extracellular matrix remodeling, and stromal signaling-may uncover additional therapeutic opportunities for cancer immunotherapy.

Macroautophagy/autophagy is a well-established homeostatic mechanism that contributes to the integrity of multiple regulatory biological activities including but not limited to the gastro-intestinal tract and cognitive integrity. Autophagy also plays a central role in tissue regeneration, metamorphosis and development whereas defects in autophagy are associated with a wide range of disorders including metabolic diseases such as diabetes, organ pathophysiologies including liver, lung and heart disease, cancer, and microbial infection. In the field of cancer therapy, most research efforts have focused on cytoprotective autophagy, with substantial preclinical and clinical studies designed to interrogate the outcomes of pharmacologically (or genetically in preclinical work) inhibiting autophagy to enhance the efficacy of chemotherapeutic agents. There is lesser but nevertheless robust evidence for the cytotoxic function of autophagy while our laboratory and a few others have identified the nonprotective form of this cellular response. However, cytostatic autophagy, a distinct functional outcome of autophagy characterized by sustained proliferative arrest, has remained relatively underexplored. Cytostatic autophagy can be defined as a cellular condition in which autophagy activation coincides with durable proliferative arrest, and in which genetic or pharmacological inhibition of autophagy relieves the growth-arrest phenotype without inducing overt cytotoxicity. In this review, we provide the first comprehensive synthesis of the scientific literature addressing cytostatic autophagy, tracing its historical development and consolidating the experimental evidence that led to its current conceptual definition. We further discuss the molecular mechanisms underlying cytostatic autophagy, including the selective degradation of key cell-cycle regulators and the interplay between autophagy and senescence-associated signaling pathways.

Poly(ADP-ribose) polymerase inhibitors (PARPi) exploit synthetic lethality in homologous recombination-deficient (HRD) cancers by trapping PARP1 on DNA, causing replication fork collapse, DNA double-strand breaks, and ultimately cell death. However, primary and acquired resistance to PARPi remains a major clinical challenge. Here, we describe a previously unrecognized mechanism for the resolution of cytotoxic trapped PARP1 through TEX264-mediated nucleophagy. We identify the p97-TEX264-nucleophagy axis as a critical pathway for the clearance of trapped PARP1 and a promising therapeutic target for overcoming PARPi resistance in HRD cancers.

Thymic central tolerance is crucial for preventing autoimmunity, but its contribution to tumor immune evasion remains poorly understood. Here, we demonstrate that plasmacytoid dendritic cells (pDCs) in the thymus have two distinct subsets, accumulating in the thymus of tumor-bearing mice, contributing to immune tolerance through clonal deletion of tumor-specific T cells and reducing newly generated T cells. Mechanistically, common dendritic cell progenitor-derived pDCs (CDP-pDCs) capture tumor antigens and migrate to the thymus in a CCR9-dependent manner, where they present these antigens to induce clonal deletion of tumor-specific T cells. Concurrently, tumor progression inhibits T cell generation by promoting the accumulation of common lymphoid progenitor-derived pDCs (CLP-pDCs) within the thymus, which further produce type I interferon to alter thymic function. CCR9 deficiency prevents thymic accumulation of both pDCs, enhancing antitumor immunity and reducing tumor growth. Our findings reveal a previously unrecognized mechanism by which tumors hijack the physiological system to establish central tolerance against peripheral antigens, thereby promoting tolerance against themselves.

Carcinoembryonic antigen (CEA) detection is crucial for early cancer diagnosis and prognostic monitoring. However, traditional CEA detection is mainly based on blood samples, which is invasive and less conducive to point-of-care testing (POCT). Therefore, in this study, we developed a flexible paper-based microneedle (MN) array electrochemical sensor, integrated with portable electronics and a mobile application, to detect CEA in interstitial fluid (ISF). The MN array was fabricated from a composite of UV-curable biocompatible resin and microcrystalline cellulose on a paper substrate impregnated with flexible UV-curable resin. The MN array was functionalized as an electrochemical sensor to detect CEA by modification with conductive ink, rGO/Thi/AuNPs nanocomposites and anti-CEA antibodies. The performance of the MN array electrochemical sensor was validated through in vitro and in vivo experiments, and the sensor demonstrated high sensitivity, good selectivity, broad linear ranges, robust stability, and strong concordance with reference methods. This study provides a new tool for CEA monitoring and offers new insights into the development of minimally invasive biomarker detection devices for POCT.

Cancer arises from genetic and epigenetic alterations that reshape chromatin, transcriptional regulation, and malignant cell states. To chart cancer-intrinsic regulatory programs, we build a pan-cancer single-cell atlas of 60 cancer cell lines spanning 16 tissue origins and 20 cancer types, comprising 240,957 snRNA-seq and 223,347 snATAC-seq profiles. Integrative analyses reveal cell-state heterogeneity, core gene-regulatory networks, and a conserved EMT axis transcending tissue of origin; copy-number analysis identifies transcription factor amplification and hyperactivation as drivers of state reprogramming. Comparing cutaneous melanoma with acral melanoma, a rare subtype underrepresented in previous studies, uncovers a universal inflammation-suppressive program in acral and an inflamed landscape in cutaneous melanoma, with JAK-STAT activity as the central discriminator. Integrating data across models and patient cohorts links tumor-intrinsic regulation to microenvironmental composition and therapeutic response. By profiling rare alongside common subtypes, this atlas offers a resource for mapping pan-cancer and subtype-specific regulatory programs shaping cell-state plasticity.

R loop homeostasis is critical for DNA double-strand break (DSB) repair; however, how R loops are resolved in this context is poorly understood. Here, we define HELZ as a unique RNA-DNA helicase that resolves R loops to facilitate homologous recombination (HR) repair. From a synthetic lethal etoposide resistance siRNA screen, we found that HELZ depletion causes R loop-mediated hypersensitivity to DSB-inducing agents, and HELZ localizes and binds to DSBs. HELZ preferentially binds to and unwinds RNA-DNA hybrids with 5'ssRNA overhangs to promote R loop resolution genome-wide and at DSBs. Interestingly, HELZ facilitates BRCA1 recruitment to DSBs by preventing R loop accumulation, thereby promoting DNA end resection and HR to prevent R loop mediated genomic instability. In summary, we show that HELZ resolves R loops critical for HR, thereby promoting genome stability and resistance to DSB-inducing agents.

Tumor senescence is a durable cell-cycle arrest triggered by oncogenic signalling, DNA damage and therapeutic stress. Although senescence can restrain malignant expansion, heterogeneous senescence-associated secretory programs can also promote tumor progression, immune evasion and treatment resistance. Crucially, the composition, magnitude and persistence of these secretory programs vary across cell types, microenvironmental niches and treatment phases, making binary detection of "senescent cells" insufficient for deciding whether specific populations should be eliminated, modulated or preserved. This review synthesizes the molecular determinants of functional, temporal and spatial heterogeneity in tumor senescence and consolidates them into an operational state space to support phase-aware intervention logic. It further evaluates how artificial intelligence, combined with single-cell and spatial profiling, imaging and circulating readouts, can enable state-resolved mapping, stratification and monitoring of senescence contexts, thereby generating testable therapeutic window hypotheses and guiding the development of staged precision senotherapies.

Why cancer arises, progresses, or proves fatal in some people but not others remains largely unresolved. Antibody repertoires, including autoantibodies targeting immune pathways, may shape cancer immunosurveillance. Mapping antibody landscapes across cancer-free, at-risk, and cancer-affected individuals could clarify their roles in cancer susceptibility and disease outcome.

Epigenetic regulation is intimately linked to cellular metabolism, enabling environmental and nutritional cues to shape gene expression programs through dynamic modifications of chromatin structure. This metabolism-epigenetics interface is mediated, in part, by the dependence of chromatin-modifying enzymes on key metabolites, including S-adenosylmethionine (SAM), acetyl-CoA, UDP-GlcNAc, and α-ketoglutarate, which serve as substrates or cofactors for DNA and histone modifications. Among these regulators, EZH2, the catalytic subunit of Polycomb Repressive Complex 2 (PRC2), has emerged as a key mediator linking metabolic state to epigenetic regulation by translating metabolic inputs into changes in chromatin architecture and gene expression. EZH2 governs developmental cell fate through H3K27me3-mediated gene repression and is frequently dysregulated in cancer, where it promotes dedifferentiation, tumor progression, and metabolic reprogramming. Importantly, EZH2 activity is itself modulated by cellular metabolic status through posttranslational modifications, including phosphorylation, acetylation, methylation, ubiquitination, and O-GlcNAcylation, which influence its stability, catalytic activity, and chromatin-binding capacity. These modifications are responsive to nutrient availability and signaling pathways involving glucose, SAM, NAD+, and other metabolic intermediates. Consequently, disruption of this finely tuned regulatory network can contribute to developmental abnormalities, metabolic dysfunction, and oncogenesis. In this review, we examine the molecular mechanisms governing EZH2 regulation and discuss how metabolic control of EZH2 shapes chromatin dynamics, cell fate decisions, and disease pathogenesis. Elucidating how metabolic signals modulate EZH2 activity will advance our understanding of development and disease while uncovering potential therapeutic opportunities to target metabolism-driven epigenetic dysregulation.

Liquid biopsy holds immense potential for the early detection of cancer, yet its clinical utility is hindered not by the lack of available tumor-associated biomarkers but by the inadequate sensitivity and clinical robustness of current molecular diagnostic tools. Nucleic acid-based biosensors have emerged as highly programmable platforms, enabling the detection of low-abundance cancer biomarkers such as microRNAs (miRNAs), circulating tumor DNA (ctDNA), and messenger RNAs (mRNAs) in complex biological fluids. Leveraging advances in DNA nanotechnology, CRISPR-Cas-mediated RNA sensing, and chemically engineered nucleic acid analogues, these biosensors achieve attomolar-level detection through nanoscale spatial confinement and enzyme-assisted signal amplification strategies. However, their clinical translation is hindered by biological sample variability, nonspecific amplification, probe degradation, and poor reproducibility. This review analyzes the core design principles of three major biosensor categories: functional DNA nanostructures, CRISPR-Cas-based sensing systems, and synthetic analogues (PNAs, SNAs). It elucidates their structural and enzymatic optimization mechanisms, distinguishes analytical from clinical sensitivity, and addresses key liquid biopsy challenges. Finally, it outlines promising strategies for clinical translation, including microfluidic integration, artificial intelligence-assisted data analysis, and theranostic nanostructures combining diagnosis with targeted therapy. This review provides a comprehensive theoretical and technical framework for the rational design of next-generation nucleic acid biosensors and offers critical insights to bridge the gap between nanoscale engineering innovation and clinical translation, ultimately advancing the development of minimally invasive and precise cancer theranostics in precision oncology.

p21-activated kinase 4 (PAK4), a critical effector of Ras homologous (Rho) GTP hydrolases (GTPases), promotes tumor progression and confers resistance to therapy by regulating malignant cellular phenotypes-such as proliferation, metastasis, and evasion of apoptosis-as well as by remodeling the tumor microenvironment (TME). Within tumor cells, PAK4 promotes proliferation by activating phosphoinositide 3-kinase (PI3K)/AKT and mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathways. It enhances invasive and metastatic potential through cytoskeletal reorganization mediated by phosphorylation of effector proteins such as LIM domain kinase 1 (LIMK1)/cofilin and actin nucleation-promoting factor WASL (N-WASP). Furthermore, PAK4 facilitates immune evasion by stabilizing programmed cell death ligand 1 (PD-L1) and inhibiting pyroptosis. At the TME level, PAK4 is activated by hypoxia-inducible factor 1-alpha (HIF-1α), inflammatory cytokines, and extracellular matrix (ECM) stiffness. This activation reprograms the immune landscape, induces aberrant angiogenesis, and promotes metabolic acidosis via the Warburg effect, collectively fostering an immunosuppressive and therapy-resistant niche. Small-molecule PAK4 inhibitors (e.g., KPT-9274, PF-3758309, compound 55) and degraders (e.g., CPS-021) effectively suppress tumor growth in preclinical models. Combining these agents with immune checkpoint blockade, chemotherapy, or radiotherapy reverses therapy resistance and reprograms the TME. Elevated PAK4 expression correlates significantly with poor patient prognosis and resistance to immunotherapy, positioning it as both a predictive biomarker and a promising therapeutic target. Elucidating the PAK4-TME axis provides a mechanistic foundation for developing novel combinatorial strategies targeting the tumor microenvironment.

Antiangiogenic immunotherapy represents a promising cancer treatment strategy. However, the efficacy of this combination approach is hindered by inadequate vascular normalization. While the fundamental role of canonical kinase-centric VEGFR-ERK signaling in angiogenesis is appreciated, the kinase-centric model overlooks non-kinase components of the pathway, such as adaptors, scaffolds, and other non-kinase interacting proteins, that could impact response to antiangiogenic therapy. Here, employing ERK kinase translocation reporter and CRISPRa screening, we revealed that the non-kinase SHC-SHCBP1 complex governed VEGFR2-ERK activity and was a dependency for aberrant tumor angiogenesis. Genetic knockout of Shcbp1 in mice sensitized tumors to VEGFR2 inhibition, preventing excessive angiogenesis, normalizing tumor vasculature, and reprogramming the immunosuppressive microenvironment to enhance immunotherapy efficacy. Mechanistically, ERK directly formed a complex with SHC and SHCBP1. Upon VEGF stimulation, active VEGFR2 recruited SHC to liberate the SHCBP1-ERK complex. SHCBP1 then triggered ERK hyperactivation by promoting ERK phosphorylation and NLS-importin-dependent nuclear shuttling, creating a feedforward loop to exacerbate pathological angiogenesis. Clinically, SHCBP1 overexpression negatively correlated with vascular normalization and antiangiogenic immunotherapy response in patients. High-throughput screening led to the development of MS1943, an inhibitor blocking SHCBP1-ERK nuclear transport. Combined with the VEGFR2 inhibitor rivoceranib and anti-PD1, MS1943 demonstrated antitumor activity against immunotherapy-resistant preclinical models, with favorable tolerance. These findings define a non-kinase-governed VEGFR2-ERK signaling pathway as a targetable dependency for tumor angiogenesis, offering the foundation for alternative antiangiogenic immunotherapy strategies.

In situ cancer vaccination, also termed intratumoral immunotherapy, transforms the tumor microenvironment into an endogenous vaccine platform by leveraging the tumor itself as a source of antigens. Unlike conventional tumor-associated antigen (TAA) or personalized neoantigen vaccines that require predefined targets and complex manufacturing, in situ cancer vaccination presents the tumor's full antigenic repertoire, including TAAs, neoantigens, post-translationally modified epitopes, cryptic peptides, and viral antigens within their native context. This broad antigen exposure elicits robust polyclonal cytotoxic T-cell responses, facilitates epitope spreading, and reduces immune escape driven by tumor heterogeneity. The therapeutic efficacy of this approach arises from the coordinated activation of multiple immune mechanisms. Programmed cell death pathways, including immunogenic apoptosis, pyroptosis, necroptosis, and ferroptosis, release tumor antigens and danger-associated molecular patterns (DAMPs) that promote dendritic-cell activation, efficient cross-presentation, and the priming of durable effector and memory T cells. The incorporation of potent adjuvants and advanced delivery platforms enhances innate-adaptive crosstalk and helps remodel the immunosuppressive tumor microenvironment. Despite these advantages, clinical translation is limited by inconsistent induction of immunogenic cell death, suboptimal intratumoral retention of therapeutics, and barriers to T-cell infiltration. Recent advances in nanomedicine-enabled delivery systems, microenvironmental modulation, and combinatorial strategies, particularly with immune checkpoint blockade, are overcoming these challenges. Collectively, these innovations position in situ cancer vaccination as a patient-tailored, broadly applicable immunotherapy capable of eliciting durable and systemic antitumor immunity.

Cellular plasticity refers to the ability of healthy cells to shift between phenotypic states and modify their characteristics to maintain tissue homeostasis and integrity. In the tumor context, cancer stem cells (CSCs) exploit this flexibility to withstand stress, facilitate tumor dissemination, and evade therapeutic interventions. Epigenetic regulation, particularly DNA methylation at CpG sites, is recognized as a well-known driver of tumor plasticity by repressing differentiation programs through modulation of chromatin accessibility. More recently, RNA modifications (epitranscriptomics) have emerged as crucial post-transcriptional regulators of gene expression that shape RNA fate and function. Among these, N6-methyladenosine (m6A), 5-methylcytosine (m5C), N1-methyladenosine (m1A), and N7-methylguanosine (m7G) contribute to the regulation of cell identity by modulating stemness-differentiation balance, stress adaptation, and epithelial-to-mesenchymal transition (EMT). Notably, dysregulation of both DNA and RNA methylation signatures is frequently observed in tumors, suggesting potential functional interactions between these regulatory layers. Emerging evidence indicates that DNA CpG methylation and RNA methylation pathways may cooperate to influence stemness, survival, and EMT-associated signaling, thereby supporting CSCs' plasticity. Although the molecular mechanisms underlying this crosstalk remain incompletely understood, accumulating studies suggest that DNA and RNA methylation could converge within interconnected regulatory networks that contribute to the control of cancer cell identity. A deeper understanding of these interactions may uncover novel vulnerabilities for targeting tumor plasticity. In this review, we summarize the current knowledge on the interplay between DNA and RNA methylation in regulating tumor plasticity, highlighting emerging mechanistic insights, functional interactions, and potential implications for future epigenetic and epitranscriptomic therapeutic strategies.

More than 2,700 human mRNA 3' UTRs have hundreds of highly conserved nucleotides, but their biological roles are unclear. These mRNAs encode proteins strongly enriched for long intrinsically disordered regions (IDRs) with hydrophobic amino acid clusters. For MYC, UTX, and JMJD3, we show that their mRNA 3' UTRs control protein activity. Rather than affecting protein abundance or localization, we find that the KDM6B 3' UTR co-translationally changes the folding of JMJD3 protein. It promotes IDR-IDR interactions and suppresses folding between domains, suggesting that RNA has IDR chaperone activity that prevents interference between hydrophobic clusters in the IDR with folding of the structured domain. 3' UTRs with chaperone activity are multivalent and mesh-like condensate-enriched, indicating the presence of localized folding environments for IDR-containing proteins. We show here that the protein sequence is insufficient for the biogenesis of fully active IDR-containing transcriptional regulators in cells, suggesting that mRNA 3' UTRs control their activity by preventing co-translational misfolding.

Spatial omics has advanced our understanding of tissue-level biology, yet tools to systematically link gene functional perturbations to spatial phenotypes and signaling pathways remain limited. To address this, we developed spatial CRISPR screen sequencing (SPAC-seq), a high-throughput spatial CRISPR screen platform, and TARDIS (target prioritization toolkit for perturbation data in spatial omics), a statistical spatial perturbation analysis toolkit. Using SPAC-seq and TARDIS, we linked gene perturbations to spatial phenotypes and pathways, uncovering how Icam1 loss in tumor cells promotes metastasis via immune suppression and macrophage polarization. In CD8+ T cells, we revealed Cd44's role in regulating spatial phenotypes by interacting with Spp1 on macrophages. We also demonstrated the model of the transcription factor-chemokine receptor axis coupling cell states with chemotaxis. SPAC-seq and TARDIS provide an effective framework to study spatially resolved functional genomics and pathways across diverse biological and disease contexts.

The mammalian genome is safeguarded within the confines of the interphase nucleus. However, genomic instability can trigger the mislocalization of nuclear DNA to the cytoplasm within micronuclei or as fragmented chromosomes. Beyond activating cell-autonomous signaling programs, whether such cytoplasmic DNA can elicit non-cell-autonomous consequences to nearby cells remains unclear. Here, we show that cytoplasmic DNAs undergo intercellular transfer through contact-dependent, cytoskeleton-based nanotube structures connecting adjacent human cells. Diverse sources of genomic instability-including exposure to mitotic spindle poisons, ionizing radiation, and Cas9-induced chromosome breakage-promote nanotube-mediated DNA transfer in both cancerous and non-cancerous cells. Transferred DNA fragments are stably inherited as functional extrachromosomal genetic elements in the recipient host genome, thereby conferring heritable phenotypic traits to the recipient cell. Our findings uncover a horizontal gene transfer-like mechanism through which direct cell-cell contact can propagate genomic instability and reshape mammalian genomes.

Naïve T cells maintain a delicate balance between quiescence and rapid activation, which involves multiple layers of regulation beyond transcription. Here, we identify the RNA modification N6,2'-O-dimethyladenosine (m6Am) and its methyltransferase PCIF1 as critical enforcers of T cell quiescence. During CD4+ T cell activation, m6Am levels are dynamically downregulated. T-cell-specific PCIF1 knockout (cKO) mice exhibit potent tumor suppression, driven by enhanced Th1 differentiation and subsequent amplification of NK cell cytotoxicity. Mechanistically, PCIF1 represses STAT1 translation via m6Am modification of its mRNA, thereby constraining Th1 commitment. Activation-induced PCIF1 downregulation releases this translational brake, enabling rapid Th1 polarization. Crucially, we identify Suramin as a pharmacological PCIF1 inhibitor that disrupts m6Am modification, boosts Th1 responses, and suppresses tumor growth. Our findings establish the PCIF1-m6Am-STAT1 axis as a translational checkpoint governing T cell differentiation and suggest that targeting PCIF1 represents a potential strategy for tumor immunotherapy.

Many transcription factors are considered "undruggable" and challenging targets due to the absence of ligandable pockets, large swaths of intrinsically disordered regions, and rapid turnover. Here, we describe a new induced-proximity therapeutic modality, Transcriptional Repression via Active Chemical Epigenetic Reprogramming (TRACER), that enforces locus-specific transcriptional silencing by recruiting endogenous corepressor complexes to transcription factor binding sites. We developed small-molecule TRACERs that tether methyl-CpG binding domain protein 2 (MBD2), a component of the Nucleosome Remodeling and Deacetylase (NuRD) complex, to transcription factor-directed ligands. Recruitment of the NuRD complex by an estrogen receptor (ER) TRACER potently suppressed ER transcriptional activity in breast cancer cells, downregulated ER target genes, and required MBD2 and histone deacetylase (HDAC1/2) for activity, confirming on-target epigenetic repression. Extending this approach to prostate cancer, an androgen receptor (AR) TRACER transcriptionally repressed both full-length AR and the drug-resistant truncation variant, AR-V7, thereby achieving >90% inhibition of AR-dependent transcription in androgen-independent prostate cancer cells with locus-specific gene repression. Collectively, these findings establish TRACERs as a generalizable modality to pharmacologically silence transcription factors through targeted epigenetic reprogramming, offering a powerful strategy for treating cancers refractory to existing therapies.

Transcriptomic perturbation profiles from tumor cell lines serve as the core molecular basis for cancer drug discovery and mechanism of action (MOA) analysis. Traditional Chinese medicine (TCM) holds great anticancer potential, yet the multi-component and multi-target properties pose major challenges for systematic mechanistic investigation. The scarcity of herbal intervention transcriptomic data severely restricts transcriptome-based anticancer TCM research, unlike widely available large-scale chemical compound perturbational datasets. This study aims to establish a predictive framework for herbal transcriptional responses in tumor cell models to address this critical data bottleneck. A transfer learning-based encoder-decoder prediction framework integrated with a self-attention mechanism was developed. The model was pre-trained on large-scale connectivity map compound perturbation datasets with paired baseline transcriptomic profiles, then fine-tuned with limited herbal perturbation data covering 11 herbs across 4 tumor cell lines using a shared gene set as the molecular basis. The model achieved strong predictive performance (mean squared error = 0.1395, R2 = 0.8561, Pearson correlation coefficient = 0.9258), outperforming baseline models with robust generalization to unseen herbal interventions. Transfer learning markedly improved prediction accuracy and stability under data-limited conditions. This framework provides a scalable, cost-effective computational approach for anticancer herbal in silico screening, preliminary MOA exploration, and multi-herb prescription synergistic pattern analysis in cancer drug discovery.

Human cancers are heterogeneous1. Dissecting how germline genetic variation and environmental factors shape tumour evolution using human datasets is limited by inherent diversity in genetic backgrounds2 and environmental exposures3-5. Here, to overcome these limitations, we re-ran early tumour evolution hundreds of times in diverged inbred mouse strains, generating matched histology and whole-genome and transcriptome sequences. The sex, environment and carcinogenic exposures were all controlled, and the study design allowed us to capture genetic variation comparable with that observed across human populations while exploiting the nested hierarchical structure of strain-litter-animal-tumour relationships. Our analyses reveal that epistatic interactions between genetic background and acquired somatic mutations result in population-specific disease progression, including choice of driver mutations, occurrence of whole-genome duplication and subclonal selection dynamics that mirror both cancer susceptibility and tumour growth rate. Even modest genetic divergence, comparable with that found across human ancestry groups, can strikingly alter selection pressures during cancer development to shape both cancer risk and the trajectory of tumour evolution.

Homotypic targeting is the inherent ability of cells to preferentially interact with cells of the same type, a phenomenon seen in cell adhesion, tissue formation and immune responses. However, its potential remains underexploited. Here we report a strategy to substantially enhance homotypic targeting through extracellular vesicles secreted by cells. By engineering the surface of small extracellular vesicles (sEVs) with lanthanides, we amplify specific cell-sEV interactions by more than 25-fold, enabling the selective capture of sEVs by cells of the same lineage even in the presence of excess off-target sEVs. We term this effect 'super homotypic targeting'. Super homotypic targeting provides a means to distinguish sEVs of different origins within highly heterogeneous sEV populations and enables two applications: using cells to detect specific sEVs and using sEVs to detect specific cells, specifically demonstrated here in the context of cancer detection from blood samples. Super homotypic targeting could hold potential for diagnostics, immunotherapy, drug delivery, rejuvenation and tissue engineering.

Proteolysis-targeting chimeras (PROTACs) offer a powerful strategy for targeted protein degradation but are frequently constrained by synthetic complexity and overreliance on a single-E3 ligase, leading to diminished efficacy in tumors with low or heterogeneous ligase expression. Here, we report a genetically encoded artificial self-splicing RNA (asRNA) platform that autonomously generates twin PROTAC effectors from a single transcript, enabling the coordinated recruitment of two distinct E3 ligases. Following transcription, the asRNA undergoes precise ribozyme-mediated self-cleavage to produce a peptide-based PROTAC that recruits the VHL ligase and an RNA aptamer-based PROTAC that engages the Dzip3 ligase, thereby activating complementary ubiquitination pathways. This dual-ligase architecture markedly enhances degradation of otherwise refractory oncogenic targets, including c-MYC and EGFR, resulting in pronounced antitumor efficacy in cancer models. The modular and programmable genetic design enables rapid reconfiguration of target-recognition elements without complex chemical synthesis. As the first single-transcript system to achieve coordinated dual-E3 ligase recruitment, this work establishes a new genetically encoded strategy for multivalent protein degradation and expands the scope of targets accessible to PROTAC-based strategies.

Clinical efficacy with chimeric antigen receptor (CAR) T cells is currently limited by numerous factors including poor initial product phenotypes and lack of engagement of endogenous immunity. Vasoactive intestinal peptide (VIP) is an immunosuppressive neuropeptide, and the antagonism of its receptor (VIPR) on T cells potentiates T cell activation. We demonstrated that VIP suppresses CAR T cell function and engineered CAR T cells to secrete a short peptide drug that antagonizes VIPR (CAR/VIPRa). Armored CAR/VIPRa T cells maintained a memory phenotype and were metabolically quiescent after manufacturing yet mounted a strong bioenergetic response after antigen stimulation. Moreover, CAR/VIPRa T cells potentiated endogenous antitumor immunity through the recruitment of host T cells. In syngeneic and xenogeneic mouse models of hematological and solid tumors, CAR/VIPRa T cells exhibited greater tumor infiltration and maintained a less exhausted memory phenotype, resulting in superior antitumor efficacy. Together, these data show that VIPRa peptides produced by armored CAR T cells can enhance T cell function and boost endogenous immunity, thereby improving tumor control.

Serine is a critical nutrient for cancer cell proliferation due to its numerous downstream biosynthetic functions. Given the ongoing clinical efforts to utilize dietary serine starvation as a potential therapy for serine auxotrophic tumors, it is important to understand how cancer cells respond to serine starvation. Here, we demonstrated that serine starvation induces dramatic changes in mRNA splicing. These effects are due, in part, to reduced translation of serine-rich proteins, including the serine/arginine-rich splicing factor (SRSF) proteins that are known regulators of mRNA splicing. Indeed, translation of SRSF6 was reduced upon serine starvation in serine-auxotrophic cancer cells and contributes to the RNA splicing changes seen upon serine deprivation. Further, reduced SRSF6 impacted the DNA damage response and cell survival in the absence of exogenous serine, and modulators of RNA splicing combined with dietary serine starvation to inhibit tumor growth. Collectively, this work describes a fundamental role for serine in supporting mRNA splicing, which may have implications in the clinical efforts to bring dietary serine starvation to the clinic.

ConspectusBoron neutron capture therapy (BNCT) is a radiotherapeutic modality in which biological selectivity is dictated not by targeting alone but by the spatial control of nuclear reactions. Following tumor-selective accumulation of 10B, neutron irradiation induces the 10B(n,α)7Li reaction, generating high-linear energy transfer particles with micrometer path lengths that confine cytotoxicity to boron-enriched cells. Despite its intrinsic precision, the clinical translation of BNCT has long been hindered by a fundamental chemical challenge: the inability to achieve sufficient, homogeneous, and durable intratumoral boron distribution. In this Account, we argue that BNCT must be reframed as a problem of chemical architecture across length scales, rather than a simple paradigm of boron delivery followed by irradiation. Building on foundational contributions from Jin-Ho Choy (layered double hydroxide-based confinement and ion-exchange systems), Hiroyuki Nakamura (molecular boron design enabling targeting and functional integration), and Koji Ono (clinical dosimetry and translational constraints), we outline how nanohybrid platforms transform BNCT into an integrated systems therapy in which materials design, biological transport, and radiation physics are co-optimized. We identify three governing design principles for next-generation BNCT. First, structural confinement and high-density boron loading enabled by layered inorganic hosts, carborane clusters, and hybrid nanocarriers, ensure sufficient payload delivery at the cellular and subcellular levels. Second, spatiotemporal control of boron distribution, achieved through surface engineering, targeting ligands, and microenvironment-responsive release, addresses the critical challenge of intratumoral heterogeneity. Third, integration with imaging and dosimetry establishes a "measure-what-you-treat" paradigm, linking molecular design directly to therapeutic outcome. We further highlight that BNCT efficacy is governed not by bulk tumor boron concentration alone but by microdistribution and subcellular localization, reflecting the short path length of high-linear energy transfer(LET) particles. This insight shifts design strategies from maximizing systemic delivery toward controlling intracellular fate and spatial precision. In parallel, the transition to accelerator-based neutron sources imposes additional constraints on boron chemistry, necessitating higher delivery efficiency and tighter coordination between pharmacokinetics and irradiation conditions. Together, these advances position BNCT as a chemically programmable precision radiotherapy, in which therapeutic outcome is dictated by the hierarchical organization of boron from molecular to nanoscale to clinical dimensions. Future progress will depend on scalable chemical design, quantitative imaging of boron distribution, and integration with emerging therapeutic modalities. We propose that the next phase of BNCT will be defined by the convergence of nanochemistry, systems-level transport control, and clinically informed design, enabling broader translation into refractory and heterogeneous cancers.

Cancer immunotherapy, exemplified by immune checkpoint blockade (ICB), remains strongly influenced by the pre-existing immune organization of the tumor microenvironment. Tertiary lymphoid structures (TLSs) are ectopic lymphoid aggregates whose density, maturation state, and spatial localization correlate with clinical prognosis and ICB response across several malignancies. Radiotherapy (RT) can reshape this immune context in opposing directions. By inducing immunogenic cell death, antigen release, cGAS-STING/type I interferon signaling, vascular remodeling, and lymphocyte recruitment, RT can create conditions that support TLS-associated antitumor immunity in selected settings. Conversely, high-dose or large-volume irradiation, poorly timed nodal exposure, and collateral injury to lymphocytes, tumor-draining lymph nodes, stromal scaffolds, and high endothelial venules can disrupt established TLSs or prevent their maturation. This review summarizes current evidence on the bidirectional relationship between RT and TLS biology, differentiating validated mechanisms from indirect evidence and hypothesis-generating translational concepts. We expound on how dose, fractionation, timing, irradiated volume, nodal management and radiation modality may influence antigen presentation, lymphocyte availability, and local immune architecture. Potential strategies such as lymph-node-aware planning, proton or heavy-ion therapy, FLASH RT, vascular normalization, STING or LTβR agonism, and ICB combinations are evaluated as investigational approaches rather than established TLS-directed clinical interventions. Future studies should evaluate whether optimized RT regimens can preserve or promote the functional maturation of TLSs, integrating paired tissue biopsies, spatial transcriptomics, advanced imaging, and circulating biomarkers to definitively correlate TLS remodeling with clinical outcomes.

Despite advancements in therapeutic cancer vaccines, clinical translation has been hindered by limited efficacy, with Sipuleucel-T remaining the only FDA-approved therapeutic cancer vaccine to date. However, recent advances in personalized mRNA vaccines, such as Moderna's mRNA-4157 and BioNTech's autogene cevumeran, have demonstrated significant reductions in recurrence risk and improved survival across several cancer types, renewing optimism in the field. Personalized cancer vaccines leverage patient-specific tumor antigens to initiate potent and targeted immune responses. This review outlines various classes of personalized vaccines, including DNA-, mRNA-, peptide-, dendritic cell-, and whole-cell-based platforms, and examines the immunological challenges they face, such as tumor heterogeneity, immunosuppressive microenvironments, and inadequate immune memory. To address these limitations, both conventional and nanotechnology-enhanced delivery systems have been developed. Notably, nanovaccines constructed from lipid-polymer hybrids, biomimetic membranes, and stimulus-responsive materials enable codelivery of neoantigens and immunostimulatory agonists, promoting enhanced lymph node targeting, dendritic cell activation, and antigen cross-presentation. Furthermore, biomimetic formulations incorporating autologous tumor membranes preserve native antigenic diversity and allow dynamic adaptation to evolving tumors. When integrated with artificial intelligence for antigen selection and multiomics for patient stratification, these platforms accelerate vaccine design and improve precision. Combination regimens with immune checkpoint inhibitors or other agents further potentiate efficacy and promote durable antitumor immunity. Increasing clinical evidence, especially in melanoma and pancreatic cancer, underscores the potential of these strategies to induce long-term protection and reduce recurrence. Overall, next-generation personalized cancer vaccines are advancing the transition from reactive treatment to proactive, precision-controlled cancer immunotherapy.

Virtual screening (VS) is a powerful approach to exploring a vast chemical space, encompassing libraries of millions to billions of compounds. However, the low hit rates of VS require testing numerous candidates to validate true binders, followed by iterative optimization cycles, which makes experimental validation costly and time-consuming. Here, we report COMBINAUT, an automated parallel synthesis platform that generates diverse chemical scaffolds to accelerate hit validation and refinement. Using a faculty-wide collection of in-house building blocks, the system enables enumeration of over 22.9 million compounds, each designed for parallelized synthesis within 32 h using repurposed solid-phase peptide synthesis equipment. Using this platform, we performed large-scale VS targeting the allosteric pocket of the immuno-oncology target, C-C chemokine receptor 2 (CCR2). Our approach facilitated the rapid synthesis and testing of 100 VS hits spanning diverse molecular architectures. In radioligand binding assays, we successfully validated nine hits with distinct scaffolds, including completely novel CCR2 ligand chemotypes. Iterative hit-to-lead optimization using the automated workflow produced cell-active CCR2 antagonists. This work demonstrates the synergy of automated synthesis and VS, enabling the efficient exploration of chemical space and the rapid discovery of novel ligands.

Hydrogen-Deuterium eXchange Mass Spectrometry (HDX-MS) is a dynamics-sensitive structural method that has rapidly achieved widespread adoption in the biopharmaceuticals industry, owing to its ability to quickly identify binding sites and to provide a molecular mechanism of action for drug candidates. However, a central limitation of conventional HDX-MS is that it has substantially lower spatial resolution than other structural techniques, typically providing information averaged over "segments" of five amino acids or more. Here, we demonstrate a sensitive, broadly applicable method for single amino acid resolution, i.e., site-specific HDX-MS measurements. Using a set of five therapeutic candidates targeting the highly druggable cancer target WDR5, we explore the greatly enhanced analytical power that arises from site specificity, including binding mode characterization, affinity ranking, and the detection of features that are "silent" in conventional peptide-level HDX-MS experiments.

Membrane proteins participate in most cellular processes from sensing the cellular environment to regulating gene expression. The fundamental unit in eukaryotic membrane proteins is the transmembrane (TM) helix, which in addition to anchoring the protein to the lipid bilayer, often plays a central functional role. We review here advances in our understanding of the factors underpinning the thermodynamic stability of TM helices in cellular membranes. Interestingly, a class of TM sequences exists that has the ability to, when isolated as a peptide, be stable both in solution and in the membrane. This Review covers such conditional TM peptides, including their discovery in the form of the pHLIP peptide, the factors that determine their membrane insertion, and the basis for their use in cancer theranostics. We also discuss how this knowledge has been the conceptual basis for the design of sequences that partition into lipid bilayers and target membrane proteins. These peptides act as both positive and negative allosteric regulators of their membrane targets. Such TM allosteric conditional peptides (TMACs) additionally constitute tools that can advance our understanding of the activity and regulation of membrane proteins.

Reversing the immunosuppressive tumor microenvironment (ITME) is a crucial obstacle in the immunotherapy of cold tumor. The STING signaling pathway has shown promise in overcoming the ITME issue. However, reversing ITME via precise activation of the STING signaling pathway remains a significant challenge. In this work, photoresponsive nanophotocages with photoimmunogenic performance were developed for ITME-reversed synergistic photoimmunotherapy of cold tumor. In the photocage design, photoresponsive boron dipyrromethene (BODIPY) was used to link a STING agonist (DMXAA) and then prepared into water-dispersible nanoparticles. The iodinated nanophotocage showed the best photoreaction efficiency and spatiotemporal controllability among the four nanophotocages. Under light irradiation, the iodinated nanophotocages precisely released the STING agonist and triggered immunogenic cell death (ICD) via the photodynamic therapy (PDT) effect, by which the ITME was reversed to enhance the immune checkpoint inhibitor (aCTLA4) immunotherapy. The robust immune response not only achieved primary tumor eradication in 80% of the mice but also significantly inhibited the distant tumor. This work offers a photoimmunogenic photocage for light-controlled STING activation for ITME-reversed cancer photoimmunotherapy.

Ionizable lipids, widely employed as pharmaceutical excipients for gene delivery, exhibit acidity-responsive membrane-destabilizing activity. This property prompted us to hypothesize that such membrane-destabilizing activity could be harnessed and selectively confined to the acidic tumor microenvironment (aTME) to achieve tumor-selective oncolysis. Herein, we report the rational design of tumor acidity-activatable oncolytic lipid nanoparticles (aoLNPs) through programmed structural modulation of ionizable lipids. By designing and screening an ionizable lipid library with systematic variations in alkyl tail length and number, we found a clear correlation between the total tail carbon number and the pH-dependent cytotoxicity of the corresponding lipid nanoparticles. Subsequent library expansion led to the identification of aoLNPE14A6-2 as the optimal candidate, which exhibited selective cytotoxicity toward cancer cells at pH 6.8 and demonstrated potent antitumor efficacy in vivo. Mechanistic investigations revealed that aoLNPE14A6-2 selectively induces lysosomal vacuolation and endoplasmic reticulum stress under tumor acidic conditions, ultimately leading to plasma membrane rupture. Notably, aoLNPE14A6-2 did not cause obvious chronic toxicity or structural damage to normal tissues after intravenous injection. This study expands the application of ionizable lipids for selective oncolytic therapy.

Enzymes catalyze biochemical reactions in all living systems, and their subcellular localizations can profoundly influence distinct cellular functions. Yet, the physiological roles of subcellular enzyme activity remain largely unexplored, owing to the lack of tools that allow imaging of enzymatic activity with organelle-level precision in living cells. Here, we present a class of fluorogenic reporters for spatiotemporally resolved imaging of enzyme activity within specific subcellular compartments, including mitochondria (Mito), lysosomes (Lyso), endoplasmic reticulum (ER), and Golgi apparatus (Golgi). These fluorogenic reporters comprise a rhodol fluorophore whose fluorescence is quenched by an enzyme-specific substrate and a bioorthogonal cage group, with signal restoration occurring only upon sequential uncaging by an organelle-localized bioorthogonal activator and subsequent enzymatic activation. We applied this versatile strategy to visualize the activity of enzymes from different families in live cells, such as leucyl aminopeptidases (LAP), γ-glutamyl transferases (GGT), and monoamine oxidase A (MAO-A). Application to LAPs revealed distinct, compartment-dependent functions: Mito-LAP activity maintained redox homeostasis in stressed cells, favoring tumor cell survival, whereas ER-LAP activity promoted antigen presentation and immune-mediated tumor cell killing. Notably, drug profiling showed that cisplatin (CDDP) and camptothecin (CPT) concurrently amplified both Mito-LAP and ER-LAP activities, eliciting opposing influences on tumor progression. These findings also inspire the development of a mitochondria-targeted inhibitor to improve anticancer efficacy. This platform offers a genetically independent, broadly adaptable framework for dissecting compartmentalized enzymology in living cells.

Patients in the liso-cel plus ibrutinib cohort of the phase 1/2, open-label TRANSCEND CLL 004 study had relapsed/refractory chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL) and received liso-cel (50×106 [dose level (DL)1] or 100×106 [DL2] chimeric antigen receptor-positive T cells) with concurrent ibrutinib from enrollment through 90 days after liso-cel infusion or longer per investigator discretion. Primary end point was complete response/remission (CR)/CR with incomplete marrow recovery (CRi) by investigator assessment. Among 56 patients who received ibrutinib plus liso-cel (DL1, n=5; DL2, n=51), median (range) age was 64.5 years (44‒77), 98% had high-risk cytogenetics, 55% had progression on Bruton tyrosine kinase inhibitor and venetoclax failure, and median (range) number of prior therapies was 5 (1‒13). Median (range) follow-up was 24.8 months (3.1‒51.8). At DL2, CR/CRi rate was 45% (95% confidence interval [CI], 31‒60) and overall response rate was 86% (95% CI, 74‒94). Median (95% CI) duration of response was not reached (NR; 28.7‒NR) for patients with CR/CRi and 41.4 months (23.3‒NR) for all responders. Median (95% CI) progression-free survival was 31.4 months (20.1‒NR). In DL1+DL2, most common grade ≥3 treatment-emergent adverse events (TEAE) were neutropenia (52%) and anemia (41%). Cytokine release syndrome was reported in 80% of patients (grade 3, 4%; no grade 4/5), and neurological events in 41% (grade 3/4, 11%; no grade 5). Ibrutinib-related TEAEs were reported in 68% of patients (grade 3/4, 43%; no grade 5). Liso-cel plus ibrutinib demonstrated notable efficacy in patients with relapsed/refractory CLL/SLL with predictable and manageable safety. Clinicaltrials.gov: NCT03331198; NCT03435796.

Azacitidine (Aza) plus venetoclax (Ven) is standard treatment for older/unfit patients with newly diagnosed (ND) acute myeloid leukemia (AML). The approved 28-day (D) Ven schedule is associated with prolonged cytopenias, causing frequent dose reductions and cycle delays. Retrospective studies show similar efficacy and reduced toxicity with abbreviated Ven dosing, but prospective data is lacking. We conducted OPTI-AML(NCT03013998), a prospective randomized phase 2 trial comparing 28D Ven (AV28) versus 14D (AV14) with Aza (75mg/m²x7D) for C1-2 in genomically agnostic ND-AML patients ≥60 years. The primary endpoint was complete remission (CR) rate achieved at any time with two cycles of therapy. Between 2023-2025, 169 patients received AV28 (n=83) or AV14 (n=86). CR across two cycles was 49.4% (AV28) versus 43% (AV14); difference of 6.4% [90%CI:-6.1% to 19.0%], not meeting non-inferiority criteria. Patients with NPM1/ IDH2 mutations had higher CR rates with AV28 (60.9% vs. 33.3%), while CR rates were equivalent (45%) for other subgroups. Composite CR rates were 80.7% (AV28) versus 68.6% (AV14) and MRD negativity was similar (77.6% vs. 76.5%). Although AV28 had more frequent treatment interruptions, count recovery after C2, grade ≥3 adverse events and early mortality were similar. In conclusion, the study did not demonstrate non-inferiority of AV14 compared with AV28 during C1-2 in an unselected ND-AML cohort. However, as the confidence interval for the difference covered 0, CR rate for AV28 was not significantly different than AV14. Certain subgroups may benefit from prolonged Ven exposure, but these findings require validation in larger studies, especially as triplet regimens evolve.

Frontline treatments for mantle cell lymphoma were historically dictated by age and fitness for intensive chemoimmunotherapy and autologous stem cell transplantation. While yielding high response rates, these approaches are associated with treatment-related morbidity and disease relapse. The therapeutic landscape is trending toward biological precision. This review highlights the timely frontline integration of novel targeted agents that directly challenge the need for traditional consolidative transplants while de-escalating or even sparing chemotherapy. Measurable residual disease is increasingly used to identify early relapses and inform the duration and individualization of therapy. By integrating targeted and immune-engaging therapeutics with optimized maintenance to deepen responses and prolong survival, these advancements aim to maximize long-term remission while minimizing toxicity.

Venetoclax (VEN) combined with a hypomethylating agent (HMA) has become a standard-of-care frontline therapy for unfit patients with newly diagnosed acute myeloid leukemia (ND AML). The VIALE-A trial established the clinical benefit of VEN plus azacitidine (AZA), with a composite complete remission rate (CRc) of 66.4% and a median overall survival (OS) of 14.7 months. Nevertheless, remission durability remains heterogeneous, and the regimen is non-curative for many patients. Improving remission rates and long-term survival therefore remains an urgent unmet clinical need. VEN/HMA-based triplet regimens incorporating targeted agents are emerging as promising strategies in ND AML, particularly FLT3 inhibitor-based combinations reported at the 2025 American Society of Hematology (ASH) Annual Meeting. Notably, longer-term follow-up data from a study of gilteritinib plus VEN/AZA in patients with ND FLT3-mutated AML showed a median OS of 29.7 months. Additional VEN/HMA-based strategies, including combinations with menin inhibitors and other novel agents, are also being investigated. Here, we provide an overview of the latest clinical updates on VEN/HMA-based triplet regimens in ND AML presented at the 2025 ASH Annual Meeting.

Anti-thymocyte globulin (ATG) has been a standard prophylaxis for graft-versus-host disease (GVHD). However, the pharmacokinetics of ATG in vivo vary significantly, and weight-based fixed dosing may not optimize efficacy while minimizing toxicity. We investigated the clinical results of a therapeutic-drug-monitoring (TDM)-based, dose-optimized ATG strategy versus weight-based fixed dosing in haploidentical haematopoietic stem cell transplantation (NCT05166967). Patients were randomly assigned in a 1:1 ratio to receive a targeted dose of ATG or a fixed dose of 10 mg/kg. The primary endpoint was the 365-day graft-versus-host disease-free and relapse-free survival (GRFS). From January 1, 2022, to January 16, 2024, 204 patients were enrolled, with 102 patients in each group. The 365-day GRFS was higher in the targeted dose group (66.7%) than in the fixed dose group (50.0%; hazard ratio [HR], 0.666; 95% confidence interval [CI], 0.4456 to 0.9954; P = 0.048). The cumulative incidence of moderate to severe chronic GVHD at day 365 was significantly lower in the targeted dose group (9.8%; 95% CI, 5.0 to 16.5) compared with the fixed dose group (22.5%; 95% CI, 15.0 to 31.1; P = 0.026). Fewer grade 3-5 infections were reported in the targeted dose group (44.1%) than in the fixed dose group (70.6%; P < 0.001). More patients in the targeted dose group achieved optimal ATG exposure (P = 0.007) and superior CD4+ T-cell reconstitution (P = 0.002). These findings support the clinical utility of a TDM-based individualized ATG dosing strategy that balances efficacy and toxicity for GVHD prophylaxis in allogeneic stem cell transplantation. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT05166967.

Selinexor for the Treatment of Myelofibrosis. SVR35- spleen volume reduction ≥ 35%, TSS- total symptom score.

Highly multiplexed immunofluorescence imaging visualizes and quantifies protein levels at single-cell resolution in intact tissues at low cost and high scalability. Analysis of these data involves multiple steps with many method and parameter choices that must be adapted to the data and analytical objectives. There is an unmet need for a toolbox that offers flexible end-to-end coverage of the workflow. Here we present 'spatialproteomics', a Python package that addresses these challenges. Spatialproteomics enables the processing and analysis of large imaging data, including steps such as segmentation, image processing and cell-type classification, while synchronizing shared coordinates across data modalities. We demonstrate spatialproteomics on images of reactive lymph nodes and B cell non-Hodgkin lymphomas from 132 patients. We showcase an end-to-end analysis from raw images to statistical characterization of how cell type composition and spatial distribution vary across indolent and aggressive lymphomas. Furthermore, we show how spatialproteomics can process Gigapixel whole-slide images.

Germline variants influence immune checkpoint inhibitor responses, but their role in engineered immune cell therapies, such as chimeric antigen receptor T cells (CAR T cells), remains unclear. We integrated whole germline sequencing from patients with lymphoma treated with axicabtagene ciloleucel CAR T cell products in the ZUMA-1 and ZUMA-7 clinical trials with detailed biomarker and functional analyses to identify variants influencing clinical toxicity and pharmacokinetics. Putative deleterious variants in STXBP2 (syntaxin binding protein 2) were enriched among patients with toxicity in ZUMA-1, although not confirmed in ZUMA-7. Mechanistically, STXBP2-deficient or variant-expressing T cells triggered increased inflammatory cytokine production and macrophage activation. Conversely, variants in ADAMTSL3, a TGFβ (transforming growth factor-β) signaling regulator, correlated with protection from toxicity across both trials. Furthermore, variants in PTPN22, a negative regulator of T cell receptor signaling, strongly associated with enhanced CAR T cell expansion, a key determinant of efficacy. Together, these findings demonstrate that germline genetics shape the safety and activity of engineered immune cell therapies, affecting future design and patient management.

Human T-lymphotropic virus 1 (HTLV-1) integrates into host DNA, resulting in life-long infection that underlies malignancy, inflammatory disease, and early all-cause mortality. HTLV-1 subtype-C endures as an endemic infection in Central Australia, and is associated with pulmonary disease. The cellular and viral features driving HTLV-1c pathogenesis remain poorly understood. We recruited a cohort of 41 First Nations participants from Alice Springs Hospital in Central Australia. We analysed plasma biomarker sVCAM1, and three circulating CD4+ T-cell phenotypes by flow cytometry: Lung-homing (TLH, CCR4+CC49d+Integrinβ7-), regulatory T-cells (TREG, CCR4+CD49d-CD127-) and CCR4- (TCCR4-). We determined the proviral load by ddPCR. Haplotype resolved proviruses were assembled from 6 donors using single provirus amplification and long-read sequencing (SPA-ONT-seq). Analysis of CD4+ T-cell phenotypes and the proviral landscape was also performed in a humanised mouse model of HTLV-1c infection. All HTLV-1c+ participants showed expansion of chronically activated TLH cells. These cells were highly infected with HTLV-1c provirus and enriched in the lung tissue of humanised mice, confirming pulmonary trafficking. Despite extensive structural diversity in the proviral landscape, defective proviruses in participants with pulmonary disease preferentially retain virulence factor hbz. Defective proviruses were detected in the sputum of HTLV-1c+ participants. We discovered chimeric HTLV-1c:human proviruses containing internalised host DNA segments, and implicated them in modulating host gene expression. HTLV-1c likely contributes to pulmonary disease through lung-homing of chronically activated CD4+ T-cells harbouring defective proviruses that retain hbz. CD49d and hbz could represent potential therapeutic targets. Single-provirus sequencing revealed previously unrecognised structural complexity, including functional viral-host chimeras, with implications for HTLV-1 pathogenesis. This work was supported by NHMRC, Mater Foundation, ACH4, and Miller Foundation PhD Scholarship.

Interleukin (IL)-10 expressing CD19 chimeric antigen receptor (CAR) T cells (META 10-19) have demonstrated encouraging clinical activity in B-cell acute lymphoblastic leukemia, but their safety and efficacy in relapsed/refractory (R/R) diffuse large B-cell lymphoma (DLBCL) remain unknown. To evaluate the safety and efficacy of META 10-19 in patients with R/R DLBCL. This nonrandomized, phase 1 clinical trial was conducted at the First Affiliated Hospital of Zhejiang University School of Medicine. Patients were enrolled from November 16, 2023, to April 7, 2025. The data cutoff date was January 20, 2026. The data analysis was conducted on January 22, 2026. Twenty patients were screened, and 13 received a META 10-19 infusion. The median duration of follow-up was 15.6 (range, 0.7-25.5) months. Following lymphodepletion with fludarabine and cyclophosphamide, patients received META 10-19 at dose levels of 2 × 103, 5 × 103 or 2 × 104 CAR T cells/kg. The primary end points were adverse events, dose-limiting toxic effects, and objective response rate. The secondary end points included complete remission (CR) and a cellular kinetic of META 10-19. Among 13 treated patients (median age, 61 years [range, 35-74 years]; 8 men [61.5%] and 5 women [38.5%]), the objective response rate was 92.3%, including CR in 11 patients (84.6%) and partial remission in 1 patient (7.7%). One patient died before response assessment because of disease-related gastrointestinal complications. Cytokine release syndrome occurred for 12 patients (grade 1: n = 8; grade 2: n = 3; grade 3: n = 1), and immune effector cell-associated neurotoxicity syndrome occurred for 2 patients (grade 1: n = 1; grade 2: n = 1). Robust in vivo CAR T-cell expansion was observed across dose levels, with a median (range) peak expansion of 660.7 (30.7-10 562.3) cells/µL. At data cutoff, 5 patients experienced a maintained CR and 7 experienced disease relapses or progression (2 with CD19 negative relapses). The results of this nonrandomized clinical trial suggest that ultralow-dose META 10-19 demonstrated promising antitumor activity and a manageable safety profile in patients with R/R DLBCL. Further investigation in larger cohorts is warranted. ClinicalTrials.gov Identifier: NCT06120166.

Plasma cell leukemia (PCL) represents an exceptionally aggressive plasma cell malignancy defined by ≥5% circulating plasma cells in the peripheral blood of patients otherwise meeting the diagnostic criteria for multiple myeloma (MM), per the International Myeloma Working Group consensus. This ultrahigh-risk disease exhibits distinctive clinical features including frequent extramedullary involvement, severe cytopenias, hypercalcemia, renal insufficiency, and/or significantly elevated β2-microglobulin and lactate dehydrogenase levels. The molecular landscape includes high-risk cytogenetic abnormalities and mutations that promote accelerated proliferation, apoptotic resistance, immune evasion, and bone marrow microenvironmental independence through dysregulated adhesion molecule and chemokine receptor expression. Although autologous stem cell transplantation, proteasome inhibitors, immunomodulatory drugs, and monoclonal antibodies improved historical outcomes, the therapeutic paradigm continues to evolve. Novel therapeutic approaches including B-cell maturation antigen-directed therapies (bispecific antibodies and chimeric antigen receptor T-cell therapy), G protein-coupled receptor class C group 5 member D (GPRC5D)-targeted therapy, and B-cell lymphoma 2 (BCL-2) inhibition demonstrate promise in treating both primary and secondary PCL. Despite these advances, PCL remains inadequately studied, with treatment approaches predominantly extrapolated from MM trials in which patients with PCL have largely been excluded. This review synthesizes current evidence and presents illustrative clinical cases demonstrating practical treatment approaches, while highlighting critical knowledge gaps requiring dedicated prospective clinical trials to meaningfully improve outcomes in this challenging disease entity.

The benefit of extended phased (∼) HLA class I∼class III∼class II haplotypes in reducing the mortality after hematopoietic cell transplantation is unknown and requires information on functional class III variation. We identified a robust class III single-nucleotide polymorphism (SNP), rs915654, informative for mortality and relapse in 1436 patients and their haploidentical related donors through multivariable regression analysis of 26 candidate class III SNPs. Three-marker haplotypes, as defined by 1 class I locus, 1 class II locus, and rs915654 were determined in patients separately from donors. Inclusion of rs915654 into relapse and mortality models already containing patient HLA-E∼DRB1 and donor HLA-B∼DRB1 improved each model (likelihood ratio test P = .06 and P = .004, respectively, for relapse; P = .10 and P = .01, respectively, for mortality). The risks of mortality and relapse increased with decreasing numbers of favorable patient and donor markers. Retesting in an independent cohort of 1141 haploidentical transplants yielded similar results. The number of unfavorable markers additionally increased nonrelapse mortality. HLA-A∼C∼B∼DRB1∼DQB1 haplotypes were defined according to their expected numbers of favorable markers, and the theoretical utility for selecting donors was explored. In summary, extended HLA class I∼class III∼class II haplotypes influence the success of transplantation and inform the biology of the major histocompatibility complex in health and disease. The selection of haploidentical donors for future patients may be optimized with knowledge of donor HLA haplotypes.

The increasing availability of genomic and transcriptomic sequencing has uncovered diverse genomic alterations and distinct gene expression profiles driving hematologic diseases, yet a data integration and sharing platform dedicated to hematology remains lacking. We developed the American Society of Hematology (ASH) HematOmics Program (ASHOP; ashop.hematology.org), a resource for exploring somatic alterations and gene fusions, transcriptomic results, and clinical data from 5960 patients spanning B-cell precursor and T-cell acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndromes, and chronic lymphocytic leukemia. Users can explore genomic alteration landscapes and comutation patterns via lollipop and matrix plots and analyze significantly altered genes in user-defined subcohorts. Transcriptomes can be explored through interactive uniform manifold approximation and projections, clustering, differential expression, and pathway enrichment. Genomic, transcriptomic features, and clinical outcomes can be correlated in a user-driven manner or combined to precisely define study cohorts. We illustrate the following 4 use cases of ASHOP: (1) stratification of DUX4-rearranged B-cell leukemias into Early/Multipotent and Committed subgroups with distinct outcomes, (2) characterization of HOXA/HOXB expression patterns in acute myeloid leukemias, (3) correlating mutational burden with mismatch repair deficiency and mutational signatures, and (4) investigation of TP53 alteration landscape. ASHOP is an open-access resource to inform genomic and transcriptomic data interpretation for hematologic malignancies and will expand to support additional diseases and data modalities from the ASH community.

Childhood T-lineage acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy with poor prognosis. Differently from B-cell precursor ALL, T-ALL lacks effective risk stratification strategies. A recent study has integrated whole genome and whole transcriptome data to define 17 distinct molecular subtypes with prognostic significance. However, clinical translation of this knowledge remains challenging due to the complexity of interpreting high-dimensional multi-omics-based data. Here, we present clinTALL, a deep learning based multi-task pipeline for pediatric T-ALL subtype classification and treatment outcome estimation. The model integrates multimodal input data and uses a neural network architecture to generate a shared latent embedding for jointly learned multi-task prediction. The competing risk-based model was used to predict event-specific outcomes. The model was trained on a publicly available multimodal dataset comprising clinical, genomic and transcriptomic features of 1309 pediatric T-ALL samples. We observed that the transcriptomic-only model achieved superior single-modality results, with 92.2% accuracy for subtype prediction and a 65.9% concordance index (C-index) for event-free survival (EFS) in a cross-validation setup. Integrating all data modalities maintained high subtype classification accuracy (91.7%) and improved the overall concordance index for EFS estimation to 67.5%. The competing risk-based model enables accurate predictions of induction failure (C-index = 96.0%) and second malignant neoplasm (C-index = 62.1%). We validated molecular subtype predictions on an internal dataset of 120 pediatric T-ALL samples and obtained an accuracy of 81.8%. To facilitate the broad application of multi-omics based subtype prediction and treatment outcome inference, we provide clinTALL as a Docker based application, allowing for user friendly access to the tool. Together, our machine learning-based framework allows for automated, accurate subtype classification and treatment outcome inference using multimodal input data, advancing precision risk stratification for pediatric T-ALL. The full source code of clinTALL is available on GitHub ( https://github.com/UKWgenommedizin/clinTALL ).

Fc receptor-homolog 5 (FcRH5) is a membrane protein that is ubiquitously expressed on myeloma cells. Cevostamab is a first-in-class, FcRH5×CD3 bispecific antibody. GO39775 is a phase 1 dose-escalation and dose-expansion study evaluating fixed-duration cevostamab in relapsed or refractory multiple myeloma. Cevostamab was initiated with step-up dosing and continued at the target dose (TD) once every 3 weeks for 17 cycles (~12 months) unless disease progression or unacceptable toxicity occurred. Primary objectives were to evaluate safety, including determination of the maximum tolerated dose (MTD), and to identify a recommended phase 2 dose and schedule (RP2D) for cevostamab monotherapy. Secondary objectives included determining the rate of response and the duration of response (DOR). As of 24 February 2025, 324 patients had been enrolled; most were heavily pretreated (median prior lines, 6; triple-class refractory, 89.5% (290/324); prior B-cell maturation antigen (BCMA)-targeted therapy, 47.5% (154/324)). The MTD was not reached. Across all TD levels (0.15-252 mg), grade 3 or 4 adverse events (AEs) and serious AEs occurred in 59.6% (193/324) and 60.2% (195/324), respectively. Grade 5 AEs excluding disease progression occurred in 4.6% (15/324); 0.9% (3/324) were considered treatment related (hemophagocytic lymphohistiocytosis, n = 2; disseminated intravascular coagulation in the context of pseudomonal sepsis, n = 1). Objective response and very good partial response or better rates were 42.1% (136/323) and 25.1% (81/323), respectively. Median DOR was 11.2 months (95% confidence interval, 8.3, 15.6). A total of 167 patients received treatment at the 160 mg TD level (RP2D). Objective response and very good partial response or better rates were 44.3% (74/167) and 25.7% (43/167) in all patients and 60.6% (43/71) and 39.4% (28/71) in the BCMA naive, respectively. Median DOR was 10.4 months in all patients and 19.7 months in the BCMA naive, with durable responses maintained after the completion of treatment. Cytokine release syndrome was grade 1 or 2 in the 0.3/1.2/3.6/160 mg triple step-up cohort (RP2D). Cevostamab had manageable safety and induced durable remissions in late-line relapsed or refractory multiple myeloma. ClinicalTrials.gov registration: NCT03275103 .

Although first-line immunotherapy achieves remission in most patients with follicular lymphoma (FL), improved biomarkers and therapeutic strategies are required to identify and manage those who relapse. We investigated the immune microenvironment at diagnosis in uniformly treated FL patients with extended follow-up (>11 years), comparing relapsed and relapse-free cases. Transcriptomic profiling revealed enrichment of inflammatory response pathways in relapsed patients, including cytokine overexpression and upregulation of CD70, a molecule implicated in immune activation and inflammation. Multiplex immunofluorescence confirmed CD70 overexpression at diagnosis in tumor cells as well as in CD8⁺ and CD4⁺ T follicular helper (TFH) cells, correlating with inferior progression-free survival. Functional studies demonstrated that CD70⁺ tumor cells were more proliferative and induced CD70 expression in T cells, suggesting a feed-forward loop sustaining immune activation. To target these cells and enhance current CAR-T approaches, we engineered dual CD19-CD70 chimeric antigen receptor (CAR) T cells by co-transducing the CD19-CAR-T ARI-0001 with a CD27-based anti-CD70 CAR. Dual CAR-T cells exhibited enhanced in vitro cytotoxicity against patient-derived spheroids of FL, diffuse large B-cell lymphoma, and mantle cell lymphoma. In FL xenograft models, dual CAR-T treatment achieved superior disease control compared to monotargeted CAR-T cells, inducing complete tumor clearance in spleen and bone marrow. Collectively, these findings provide strong preclinical rationale for the clinical development of CD19-CD70 dual CAR-T therapy to improve outcomes in high-risk FL and potentially other B-cell non-Hodgkin lymphomas.

Early-phase clinical trials of Bruton's tyrosine kinase (BTK) degraders have demonstrated efficacy in patients with BTK inhibitor-resistant chronic lymphocytic leukemia (CLL). How clinical resistance to BTK degraders arises is unknown. Here we sequenced serial CLL samples from patients enrolled in the phase I trials of zelebrudomide and bexobrutideg and observed recurrent expansion of preexisting BTK A428D mutations at relapse. Unlike previously studied BTK inhibitor resistance mutations, BTK A428D conferred pan-resistance to BTK inhibitors and degraders. In the absence of BTK-directed therapies, however, cells bearing BTK A428D exhibited a competitive disadvantage. A crystal structure of BTK A428D revealed that the mutant aspartate clashes with the adenine ring of ATP and the adenine-mimetic moiety of BTK inhibitors and degraders. Combining BTK degraders with venetoclax mitigated the expansion of BTK A428D. These results provide the molecular basis for clinical resistance to BTK degraders and will inform the development of next-generation BTK degrader therapies.

Mantle Cell Lymphoma (MCL) is an aggressive B-cell non-Hodgkin lymphoma, with frequent relapses and shorter responses with every subsequent treatment. MCL depends on growth factors and cytokines derived from microenvironmental cells for its growth and can alter the immune system to evade recognition and subsequent elimination. The soluble factors secreted by MCL can contribute to endothelial differentiation, lymphangiogenesis, and clonal selection under hypoxic conditions, thereby evading the DNA damage response. Targeting the tumor microenvironment and angiogenesis is an active area of research and development, as the angiogenic gene signatures in MCL remain poorly understood. To address this knowledge gap, we performed transcriptomic analyses of MCL patient cohorts and identified 10 key angiogenic genes upregulated in MCL. We focused on four receptors (FGFR1, VEGFR1, VEGFR2, and PDGFRB) that have receptor tyrosine kinase activity and are localized to the plasma membranes of MCL cells. These receptors were assessed for therapeutic targeting potential in four independent preclinical models, including patient-derived xenografts, cell-derived xenografts, bone marrow-derived xenografts, and a genetically engineered murine model of MCL. Our work establishes that simultaneous targeting of multiple kinases, such as FGFR1 and VEGFR2, is a promising therapeutic strategy for patients with MCL.

Chimeric antigen receptor (CAR) T cells are highly potent therapies approved by the U.S. Food and Drug Administration (FDA) for several hematological malignancies. However, efficacy remains variable due to resistance, antigen modulation, systemic toxicities, and relapse. Developing improved CAR T cell therapies relies on preclinical models that accurately predict clinical outcomes. For this purpose, immortalized tumor cell lines, such as the Burkitt's lymphoma line Raji, are commonly used. CD19-targeting CAR (CAR19) and non-transduced (NT) human T cells were co-cultured with target cell lines in vitro to assess T cell cytotoxicity, cytokine production, activation, and proliferation. Major histocompatibility complex (MHC) class I and II blocking antibodies and TCR knockout (KO) T cells were used to validate T cell receptor interactions. In vivo, Raji wild-type (wt) or CD19 KO tumor-bearing mice were treated with CAR19 or NT T cells, and survival, tumor burden, serum cytokines, and T cell phenotype in tissues were analyzed. Statistical analyses using either paired/paired ratio/unpaired two-tailed Student's t tests, two-way analysis of variance, and log-rank (Mantel-Cox) tests were performed using GraphPad Prism 9. We report that T cells generated from multiple different donors exhibit not only strong CAR-mediated cytotoxicity against CD19-expressing Raji WT cells but also, unexpectedly, CAR-independent reactivity against Raji CD19 KO cells. This reactivity was observed both in vitro and in vivo, and was associated with pro-inflammatory cytokine secretion and upregulation of T cell activation markers, consistent with endogenous T cell receptor (TCR) engagement. NT T cells displayed similar activation, cytotoxic, and cytokine responses against Raji cells. These effects failed to occur using other B cell leukemia/lymphoma or solid tumor cell lines in vitro. Both CD4+ and CD8+ T cell populations contributed to this alloreactive response, which could be attenuated by blockade of MHC class I or II, highlighting a TCR:MHC-dependent mechanism. These findings demonstrate that certain tumor lines can elicit strong allogeneic T cell responses independent of CAR specificity, revealing an important limitation of commonly used preclinical models and highlighting the potential for false-positive assessments of CAR T cell efficacy. Furthermore, these results highlight the importance of validating tumor models for alloreactivity prior to use in preclinical testing.

Acute myeloid leukemia (AML) is an aggressive blood disorder characterized by rapid growth of poorly differentiated myeloid cells. Gain-of-function mutations in isocitrate dehydrogenases (IDHs) are detected in ~20% of AML and ~80% of secondary gliomas. Mutant IDH1/2 isoenzymes acquire neomorphic activity to produce 2-hydroxyglutarate (2-HG) oncometabolite, resulting in hypermethylated DNA and histones, altered gene expression, and blocked differentiation of hematopoietic progenitors. Here, we presented preclinical development of LY3410738, an oral, dual IDH1/2 inhibitor with potential to penetrate the blood-brain barrier. LY3410738 covalently inhibited mutated-IDH1/2, reduced 2-HG levels at low nanomolar concentrations in human AML and glioma models, and demonstrated efficacy in AML patient-derived xenografts (PDXs) in vivo, inducing myeloid differentiation. LY3410738 retained in vitro activity in cancer models with acquired secondary IDH1/2 mutations conferring resistance to ivosidenib and enasidenib. LY3410738 synergized and was well tolerated with standard-of-care regimens such as cytarabine, azacitidine, venetoclax, or midostaurin in IDH1/2-mutated AML PDXs.

Children with Down syndrome (DS) have an elevated risk of developing myeloid leukemia (ML; ML in DS [ML-DS]). In addition to mutations in GATA1, which generate the truncated isoform GATA1-short (GATA1s), ML-DS requires additional somatic gene mutations, most frequently in cohesion and Polycomb repressive complex 2 (PRC2) genes. Here, we show that PRC2 insufficiency underlies ML-DS pathogenesis. Transplantation of Gata1s fetal liver cells followed by deletion of the cohesion subunit Stag2 and/or the PRC2 component Ezh2 induced megakaryocyte-biased differentiation and expansion of megakaryocytic progenitors, culminating in lethal myelofibrosis. Mechanistically, loss of Stag2 or Ezh2 reinforced Gata1s-driven reduced chromatin accessibility at erythroid transcription factor target loci in premegakaryocyte/erythroid progenitors (pre-MegEs), thereby promoting megakaryocytic skewing. Ezh2 loss attenuated the Gata1s-mediated global elevation of H3K27 trimethylation in pre-MegEs, resulting in derepression of a broad set of PRC2 target genes and establishing a functionally PRC2-insufficient state. Similarly, Stag2 loss induced a moderate but significant degree of PRC2-insufficient state in Gata1s progenitors. Furthermore, chromosome 21-encoded miR-125b blocked megakaryocytic differentiation of Gata1s progenitors lacking either Stag2 or Ezh2 alone but drove full transformation and expansion of CD150+Sca-1+c-Kit+ leukemic stem cell-like populations only upon concurrent loss of both Stag2 and Ezh2, leading to acute megakaryoblastic leukemia in mice. These findings reveal that cohesin and PRC2 insufficiencies converge on PRC2 dysfunction while exerting distinct epigenetic effects and synergize with trisomy 21 and GATA1s to remodel the epigenetic landscape, driving progression from a preleukemic state to overt leukemia.

Innate immunity is increasingly recognized as a driver of neurodegeneration, although pathogenic mechanisms are incompletely understood. Langerhans cell histiocytosis (LCH) is an inflammatory myeloid neoplastic disorder caused by activating somatic mutations in MAPK pathway genes, most commonly BRAFV600E, in myeloid precursors. A subset of patients with LCH develop progressive neurodegeneration (LCH-ND). We generated a human induced pluripotent stem cell (iPSC) model from patients with somatic hematologic mosaicism for BRAFV600E. Brain macrophages/microglia from LCH iPSCs exhibit unique disease-specific pathogenic features. Stepwise differentiation identified hematopoietic progenitors as hyperproliferative, whereas brain macrophages were apoptosis resistant. Through application of cerebral organoids and a humanized murine xenotransplantation model, we identify marked heterogeneity of differentiation potential within clonal BRAFV600E lines in vivo. This model phenocopied human-specific phenotypes, including dense basal ganglia foci of abnormal macrophages, marked neurodegeneration with astrogliosis, and progressive ataxia. This approach will allow for preclinical testing of therapeutics for LCH-ND.

Although survival rates for acute lymphoblastic leukemia (ALL) have improved dramatically over the past 60 years, outcomes vary across different molecular subtypes and risk categories, and a significant number of patients remain difficult to treat. ROR1 is a receptor tyrosine kinase expressed in B-cell ALL, particularly in patients with TCF3::PBX1 gene rearrangements. ROR1 has served as the target of several immune-based therapies including antibody-drug conjugates (ADCs), bispecific T-cell engagers, and CAR T-cell therapies. Here, we evaluated the ROR1-targeting ADCs zilovertamab vedotin (ZV, previously known as MK-2140 or VLS-101) and VLS-211 in vivo against a panel of ALL patient-derived xenografts with variable ROR1 expression. Both agents showed modest activity, which was dependent on ROR1 expression. Notably, we identified pediatric patients with TCF3::HLF gene fusions, which is a highly chemoresistant ALL subtype, as having some of the highest ROR1 expression in pediatric ALL and show that this subtype is susceptible to targeting ROR1 via ADC-based therapy. Given that ZV has a favorable toxicity profile in patients with hematological malignancies, it may have some utility in the treatment of very high-risk pediatric B-ALL, particularly cases with TCF3::HLF gene fusions.

Graft-versus-host disease (GVHD) remains a major barrier to the success of allogeneic hematopoietic stem cell transplantation (HSCT). In preclinical models, dysregulation of IL-6 in the peri-transplant period promotes GVHD via STAT3-dependent T cell differentiation and monocyte activation but the nature of immunological effects invoked in patients remains unclear. To advance our understanding of IL-6 signaling in humans during HSCT, we performed single-cell RNA sequencing on circulating CD14+ monocytes and CD4+ T cells from patient samples within a clinical trial of IL-6R inhibition (tocilizumab (TCZ) or placebo) on a backbone of calcineurin inhibition (CNI) and short-course methotrexate. We studied patients who did not develop acute GVHD and included a placebo-treated group analyzed prior to the development of acute GVHD. IL-6R inhibition promoted Type-I IFN-associated transcriptional programs in both CD4+ T cells and CD14+ monocytes. Surprisingly, IL-6R-inhibition with TCZ profoundly enhanced cytolytic CD4+ T cell differentiation. In experimental HSCT and chimeric antigen receptor T cell systems, genetic deletion of IL-6 signaling in donor T cells enhanced the expansion of cytolytic Eomes+ CD4+ regulatory T cell subsets whilst attenuating Th1 and Th17 differentiation. Consistent with the promotion of this cytolytic CD4+ phenotype, anti-tumor effects were improved in the absence of IL-6 signaling. In summary, these data demonstrate that IL-6R inhibition during cell therapy imprints Type-I IFN programs and cytolytic CD4+ T cell differentiation, including the Eomes+ fraction associated with favorable immunotherapy outcomes.

Sickle cell disease (SCD) is a blood disorder affecting millions worldwide. Emerging evidence reveals that SCD pathophysiology increases the risk of myeloid malignancies and hematopoietic stem cell (HSC) dysfunction, likely because of chronic stress on bone marrow. To investigate this further, we interrogated bone marrow hematopoietic stem and progenitor cells (HSPCs) from mice and individuals with SCD and observed molecular signatures of chronic cellular stress including oxidative stress, DNA damage, and hallmarks of senescence. Consistent with these findings, SCD HSPCs displayed transcriptomic dysregulation of senescence-associated molecular programs and diminished mitogen response with prolonged cell cycle kinetics during time-lapse live cell imaging. SCD mice displayed a marked loss of immunophenotypic bone marrow HSPCs by flow cytometry and functional blood repopulating HSPCs in transplantation studies, whereas human SCD bone marrow HSPCs exhibited poor ex vivo hematopoietic colony-forming ability, and these phenotypes were reversed after senescence-targeting therapy with either ABT-263 (navitoclax) or the combination of dasatinib and quercetin. Thus, treatment with senescence-targeting therapy improves bone marrow HSPC function in vivo in mice and ex vivo in cells from individuals with SCD and could represent a possible strategy to improve HSPC health, promote manufacture of high-quality bespoke clinical products, and potentially enhance the safety of potentially curative gene therapies using autologous HSPCs from individuals with SCD.

Human T-cell leukemia virus type 1 (HTLV-1) is the causative agent of adult T-cell leukemia/lymphoma (ATLL) and the neuroinflammatory disease, HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The HTLV-1 Tax regulatory protein plays a critical role in HTLV-1 persistence and pathogenesis; however, the underlying mechanisms are poorly understood. Here we show that Tax dynamically regulates mitochondrial reactive oxygen species (ROS) and membrane potential to trigger mitochondrial dysfunction. Tax is recruited to damaged mitochondria through its interaction with the IKK regulatory subunit IKBKG/NEMO and directly engages the ubiquitin-dependent PINK1-PRKN/parkin pathway to induce mitophagy. Tax also recruits autophagy receptors CALCOCO2/NDP52 and SQSTM1/p62 to damaged mitochondria to induce mitophagy. Furthermore, Tax requires PRKN to limit the extent of CGAS-STING1 activation and suppress type I interferon (IFN) induction. HTLV-1-transformed T-cell lines and PBMCs from HAM/TSP patients exhibit hallmarks of chronic mitophagy, and inhibition of PRKN in HTLV-1-transformed cell lines downregulates p19 Gag expression and induces cell death. Collectively, our findings suggest that Tax manipulation of the PINK1-PRKN mitophagy pathway represents a new HTLV-1 immune evasion strategy important for maintaining viral gene expression and cell survival.

Accurate risk stratification in stage II colorectal cancer is essential for treatment decision-making, as current guidelines recommend adjuvant chemotherapy only for patients with a high-risk of relapse. We aimed to develop and validate an AI-based approach for automated invasive front assessment to improve prognostic stratification in this population. We developed SÉMIL (Semantically-Enhanced Multiple Instance Learning), integrating vision-language foundation models with attention-based multiple instance learning for automated invasiveness assessment from H&E-stained whole slide images. We trained and validated SÉMIL on 1,608 H&E-stained WSIs from three cohorts (Austin n=697, MCO n=478, DYNAMIC n=433). We compared SÉMIL performance against manual pathologist assessment and non-semantic MIL approaches. For binary classification, SÉMIL outperformed non-semantic MIL across all cohorts (external validation: AUC 0.713-0.821 vs 0.686-0.803). For survival prediction, SÉMIL demonstrated validated prognostic stratification in both the internal (Austin: HR=4.73, p=0.0012) and the two external (MCO: HR=2.84, p=0.0032; DYNAMIC: HR=2.10, p=0.0396) stage II validation cohorts. Critically, among National Comprehensive Cancer Network (NCCN) guideline-defined high-risk stage II patients, SÉMIL successfully stratified outcomes across all three cohorts (HRs 2.96-3.50, all p<0.05), demonstrating consistent reproducible performance. In multivariate analysis of the combined stage II cohort (n=1,220), SÉMIL retained independent prognostic significance (HR=1.98, p=0.005) after adjusting for conventional clinicopathological features including T stage, MMR status, and lymph node examination adequacy. Concordance analysis between SÉMIL and manual assessment showed concordant infiltrative classification identified the highest-risk group (HR=3.96, p<0.0001), with discordant cases showing intermediate risk. SÉMIL demonstrates validated prognostic stratification in stage II colorectal cancer, with potential utility for refining risk assessment within NCCN guideline-defined high-risk categories where treatment decisions are most challenging.

Tumour cells and tumour-associated stroma are key components of the tumour microenvironment, and their interaction impacts disease progression and treatment resistance in rectal cancer. This study introduces a computational approach to quantify tumour cell density (TCD) within epithelial and stromal regions and assess whether treatment response differs according to TCD status in patients with locally advanced rectal cancer (LARC) undergoing neoadjuvant chemoradiotherapy (nCRT). The data of 414 ARISTOTLE trial (ISRCTN09351447) participants with available digitised pre-treatment biopsies were analysed in this study. We defined TCD as the proportion of tumour cells within the tumour and stroma tissues and quantified TCD using an AI framework applied to digitised haematoxylin and eosin-stained whole-slide images. The patients were stratified as TCD-high/TCD-low using a cut-off value of 0.5 (50% of tumour cells). The TCD status was combined with treatment arms [CRT (capecitabine + radiotherapy) and IrCRT (experimental capecitabine + irinotecan + radiotherapy)] to stratify the disease-free survival (DFS), overall survival (OS) and pathological complete response (pCR) rates. Among the patients analysed, 188 (45%) of 414 patients were classified as TCD-high and 226 (55%) as TCD-low. A significant treatment-TCD interaction was observed for both DFS (χ2 = 6.88, p = 0.009) and OS (χ2 = 10.61, p = 0.001). In the TCD-high subgroup, the use of IrCRT was associated with significantly longer DFS (HR = 0.57, 95% CI: 0.36-0.90, p = 0.014) and OS (HR = 0.50, 95% CI: 0.30-0.84, p = 0.008), along with a higher pCR rate (20 [23%] vs. 8 [11%], OR = 2.46, 95% CI: 1.01-5.98; p = 0.042 [p = 0.13 after adjustment for multiple testing]), compared to CRT. However, in the TCD-low subgroup, no significant difference in DFS was observed between treatment arms (HR = 1.29, 95% CI: 0.85-1.95, p = 0.22), while patients who received IrCRT exhibited a trend towards worse OS (HR = 1.55, 95% CI: 0.96-2.51, p = 0.07). No significant difference in pCR rates was observed (12 [14%] vs. 23 [22%], OR = 0.60, 95% CI: 0.28-1.29; p = 0.19). In this post-hoc, hypothesis-generating analysis of the ARISTOTLE trial, higher TCD was associated with differential outcomes after irinotecan-intensified neoadjuvant chemoradiotherapy compared with standard chemoradiotherapy. These findings support further evaluation of AI-derived TCD as a candidate predictive biomarker in independent retrospective and prospective cohorts. Cancer Research UK Radiation Research Network - Project Seed Funding (RRNPSF-Jan21/100001), Cancer Research UK ARISTOTLE sample collection grant (A18745), UK Research and Innovation Future Leadership Fellowship (No. MR/T040785/1) and the Radiation Research Unit at the Cancer Research UK City of London Centre Award (C7893/A2899).

Neoplastic recurrence risk at scheduled surveillance intervals after colorectal endoscopic submucosal dissection (ESD), as well as its association with histologic risk features, is unclear, resulting in uncertainty for post-ESD surveillance recommendations. We conducted a systematic review and meta-analysis of 1-, 3-, and 5-year post-ESD neoplastic recurrence with subgroup analyses. A systematic search was performed up until October 2025. Eligible studies were assessed for neoplastic recurrence at or near the resection site at 1-, 3-, and 5-year scheduled surveillance colonoscopies. Rates of metachronous neoplastic lesions at the same time-intervals were also collected. Data were pooled using a random-effects model, and subgroup analyses were conducted for histology of the index lesion (low or high grade dysplasia, and T1 cancer) and resection quality (R0 vs. non-R0). 10 studies encompassing 5306 lesions met the inclusion criteria. The pooled R0 resection rate was 82% (95%CI 79%-85%). Overall neoplastic recurrence rates were low and stable over time: 1.2% (95%CI 0.4%-2.6%), 1.4% (95%CI 1.0%-1.9%), and 1.9% (95%CI 0.9%-3.1%) at 1, 3, and 5 years, respectively. Malignant recurrence was rare (0.2%) and confined to noncurative resections or deep submucosal invasion. The pooled rate of metachronous lesions was 1.4% (95%CI 0.4%-3.0%). Limited time-stratified data suggest progressive accrual of metachronous neoplasia during follow-up rather than early post-ESD failure. Neoplastic recurrence is rare up to the 5-year endoscopic surveillance, especially in those with R0 resection and favorable histology. In low risk patients, an extended surveillance interval of up to 5 years can be considered, mostly because of an increased risk of metachronous lesions.

Deficient DNA mismatch repair/microsatellite instability-high (dMMR/MSI-H) cancers are very sensitive to immune checkpoint inhibitors (ICIs), yet their use is frequently complicated by immune-related adverse events (irAEs). Our study analyzed the impact of grade ≥3 irAEs (immune-related severe adverse events (irSAEs)) on survival in patients with dMMR/MSI-H digestive cancers treated with ICIs. We conducted an international, multicenter, ambispective study involving 1,175 patients from 34 centers. The primary endpoint was the correlation between the occurrence of irSAEs and progression-free survival (PFS). Secondary endpoints included factors associated with irSAEs, correlation between irSAEs and overall response rate (ORR) and overall survival (OS). Prespecified landmark and time-dependent survival models were used to account for the time to irSAE occurrence. Among 1,175 patients treated with ICIs for digestive cancers, 49.1% were female, median age was 66.9 years (IQR 53.7-77.3) and 82.9% had colorectal cancer. Overall, 382 patients (32.5%) had an irAE, including 117 (10.0%) irSAEs. Median time to irSAEs occurrence was 3.78 months (IQR 1.97-7.62). The most frequent irSAEs were gastrointestinal (3.4%), hepatic (1.8%) and dermatologic (1.0%). The only factor associated with irSAEs was the use of an ICI combination versus monotherapy (20.0% vs 11.4%, p=0.03). Among patients with irSAEs, 71.5% received oral and 29.8% intravenous corticosteroids, and 15.7% immunosuppressive agents. Overall, in 26.9% of patients, ICIs were resumed following irSAEs, with an irAE recurrence rate of 57.1%. irSAE was associated with a better ORR (48.6% vs 34.6%, p<0.0001). The multivariable Cox model revealed that irSAE as a time-dependent variable was not associated with better PFS (HR=1.280 (95% CI 0.898 to 1.824), p=0.172) or OS (HR=1.043 (95% CI 0.712 to 1.528), p=0.828). Likewise, landmark analysis at 3 months and 6 months also showed that irSAEs were not associated with better PFS or OS. The occurrence of irSAE was not independently associated with better survival in patients with dMMR/MSI-H digestive cancers treated with ICIs.

PIK3CA is mutated in ~15% of colorectal cancers (CRC). PI3K regulates immunity, inhibition potentially enhances anti-tumor immunity. We launched a phase 1/2 trial of copanlisib (PIK3CA inhibitor) and nivolumab (anti-PD-1) in metastatic microsatellite stable CRC (NCT03711058): Cohort A: PIK3CAwt (n = 17) and Cohort B: PIK3CAmut (n = 22). Copanlisib/nivolumab is well tolerated with recommended phase 2 dose of nivolumab 480 mg day 1 and copanlisib 60 mg days 1/8/15 of a 28-day cycle. Primary endpoint of objective response rate at 6 months was not met with Cohort A: 0/17 and Cohort B: 2/22 having 6-month treatment response. Secondary endpoints are median progression-free survival (Cohort A: 1.7 months; Cohort B: 1.6 months), median overall survival (Cohort A: 8.5 months; Cohort B: 6.7 months), duration of response (Cohort A: 13.1 months; Cohort B: 17 months) and 6-month disease control rate (Cohort A: 2/17; Cohort B: 3/22). Secondary endpoints were not statistically different between these cohorts.

The impact of computer-aided detection (CADe) systems on lesion detection remains uncertain. We evaluated whether a CADe system improves adenoma detection rate (ADR) in a colorectal cancer (CRC) screening program. This multicenter randomized clinical trial included patients aged 40-79 years who underwent colonoscopy after a positive fecal immunochemical test (FIT) or for surveillance. Patients were randomized to either conventional or CADe-assisted colonoscopy. The primary end point was ADR. Secondary end points were detection rates for serrated lesions (SLDR), polyps (PDR), advanced adenomas (AADR), and advanced serrated lesions (ASLDR). We performed subgroup analysis according to indication, endoscopist ADR, and bowel cleansing. Effect estimates were reported as adjusted risk ratios (aRRs) and 95%CIs. 857 patients were randomized and 827 were included in the analysis. Baseline characteristics were comparable between groups (mean age: 61.7 [SD 6.5]; male 490; positive FIT: 552). We did not find statistically significant differences between CADe-assisted and conventional colonoscopy for ADR (60.8% vs. 57.7%; aRR 1.05, 95%CI 0.94-1.17). No significant differences were observed for SLDR (8.8% vs. 7.2%; aRR 1.04, 0.65-1.64), PDR (69.6% vs. 68.8%; aRR 1.00, 0.91-1.10), AADR (23.6% vs. 24.4%; aRR 0.90, 0.71-1.14), or ASLDR (4.9% vs. 4.8%; aRR 0.93, 0.51-1.68). Subgroup analysis showed no significant differences. In colonoscopies performed in a CRC screening program, CADe-assisted colonoscopy did not increase ADR compared with conventional colonoscopy.

In the progression from inflammatory bowel disease to associated cancer, the clonal mutational landscape shifts from selection of mutations in inflammatory genes to selection for cancer-driver mutations. How prevalence and expansion of either type of mutant clones could be impacted by the cellular environments in which they arise and how this affects the neoplastic outcome of colitis remains unknown. Here we combine in vivo lineage tracing, in silico modeling, mutational profiling and spatial transcriptomics in a mouse model of colitis-associated tumorigenesis to capture clone fates associated with chronic inflammation. We identify epithelial- and immune-enriched neighborhoods and propose a model in which establishment of a reparative tissue environment facilitates tumor initiation by promoting the selection and expansion of pro-oncogenic clones, reducing the span of inflammation-resistant neighborhoods containing nononcogenic clones.

In their recent paper, Moore and colleagues demonstrate that, upon KRAS hyperactivation, colorectal cancer (CRC) growth is driven by a reprogramming of Lrg5+ intestinal stem cells progeny towards the acquisition of a regenerative phenotype. They find that this phenotype is regulated by a balance between WNT-related intestinal stem cell and MAPK-related regenerative and proliferative transcriptional programs. By targeting both pathways, they are able to suppress this dynamic plasticity and achieve tumor regression in cell line and mouse models. The antagonistic relationship between these central pathways defined here provides key insight into genomic patterns of CRC and targeted therapy strategies.

Federated Learning (FL) enables collaborative training across institutions without sharing sensitive data, a solution for privacy-preserving AI in medical imaging. However, hospital deployment remains challenging due to strict data protection regulations, heterogeneous infrastructures, and limited network accessibility behind firewalls. We introduce TheODen, an open-source framework for Federated training on histopathology Whole Slide Imaging (WSI). It requires no open client-side ports, enabling training through firewalls via a secure reverse-proxy architecture. We conducted, to our knowledge, the first nationwide FL study for histopathology segmentation of colorectal cancer across three German university hospitals, using breast and colorectal cancer datasets without opening firewall ports. TheODen achieves robust segmentation, with global average dice scores of 0.764 on BCSS and 0.754 on SemiCOL despite data heterogeneity and network constraints. These findings underline TheODen's potential to facilitate secure, large-scale collaborations between medical institutions and to accelerate clinical translation of AI models under real-world infrastructure constraints, providing a privacy-preserving-by-design architecture for future collaborations.

Therapy-resistant cancers remain largely incurable because malignant cells acquire metabolic adaptations that sustain survival under chronic cytotoxic and oxidative stress. One such adaptation is constitutively enhanced macropinocytosis, enabling aggressive cancer cells to scavenge extracellular nutrients and maintain redox homeostasis. Here, we introduce a macropinocytosis-exploiting polymer-metal strategy that converts this metabolic dependency into a lethal vulnerability. We developed PPS02, a polyaspartic acid sodium salt-based metal complex that is preferentially internalized by cancer cells via macropinocytosis, while remaining largely excluded from normal epithelial cells. This cancer-preferential uptake enables intracellular delivery of selenomethionine and ferrous iron, resulting in intracellular H2O2 accumulation, mitochondrial reactive oxygen species overload, and activation of necroptotic cell death. Macropinocytic activity was significantly elevated in patient-derived metastatic colorectal cancer cells but remained minimal in normal colonic epithelial cells, demonstrating pronounced cancer selectivity. Accordingly, PPS02 exhibited negligible cytotoxicity toward normal colonic epithelial cells while effectively suppressing the viability of both nonmetastatic and platinum-resistant metastatic colorectal cancer cells. In patient-derived xenograft models, PPS02 induced sustained tumor regression without overt systemic toxicity under the experimental conditions, whereas cisplatin failed to control metastatic tumors and caused significant adverse effects. Collectively, these findings support macropinocytosis-driven redox imbalance as a therapeutically exploitable vulnerability and demonstrate a polymer-metal platform that preferentially induces necroptosis in drug-resistant cancer while sparing normal tissues.

Obesity is a major risk factor for colorectal cancer (CRC), yet the mechanisms linking obesity-associated gut dysbiosis to tumor progression remain unclear. Here, we show that a high-fat diet and fecal microbiota from patients with obesity-associated CRC deplete the GABA-producing commensal Bacteroides ovatus, resulting in reduced luminal GABA and accelerated tumorigenesis. Microbial GABA activates epithelial GABAB receptor signaling and induces TPI1 through the PI3K-HIF1α pathway. Increased TPI1-derived glyceraldehyde-3-phosphate inhibits PPP1CA, maintains YAP phosphorylation, restricts nuclear YAP activity, and suppresses pentose phosphate pathway flux, thereby limiting tumor growth. Consistently, obesity-associated CRC exhibits reduced fecal GABA, decreased TPI1 expression, and metabolic rewiring. A GABA-deficient B. ovatus mutant fails to restore GABA or suppress tumors despite normal colonization, whereas oral GABA supplementation or recolonization with wild-type B. ovatus markedly reduces tumor burden. These findings identify a microbiota-neurotransmitter-metabolism axis linking obesity to CRC and suggest microbiota-based GABA restoration as a potential preventive strategy.

Chemical composition, oxidative stability, and biological activity of cold-pressed pomegranate (PG), blackcurrant (BC), and prickly pear (PP) seed oils were investigated. Oils were characterised for fatty acids and minor constituents, and oxidative stability was assessed by differential scanning calorimetry (DSC). Biological activity was evaluated in colon cancer (LoVo, LoVo/Dx) and non-tumorigenic (CCD 841 CoTr) cells using viability, apoptosis, ROS generation, and Rhodamine 123 accumulation assays. PG exhibited the highest PUFA content (90.0%) and the lowest lipophilic antioxidant-to-PUFA ratio (5.92), whereas BC (32.00) and PP (8.78) showed greater antioxidant balance. BC contained the highest tocopherol (1649 mg/kg) and carotenoid levels, while PG contained the highest amount of quantified phenolic and phenolic-related compounds. PP showed the greatest oxidative stability at 100 °C, whereas BC was the most stable at higher temperatures and PG was the least stable. All oils reduced cancer cell viability and induced apoptosis, with PG showing the strongest pro-oxidant activity. The results suggest that the balance between lipophilic antioxidants, fatty acid composition, and the native oil matrix influences oxidative stability and may contribute to the observed biological activity.

Early detection of colorectal polyps is crucial to reduce the morbidity and mortality associated with colorectal cancer. However, during endoscopy, continuous camera motion and the complex clinical environment often degrade key visual cues (e.g., polyp morphology, texture, and boundaries). This leads to cross-frame view shifts, which are characterized by heterogeneous appearances of the same lesion over time, thereby introducing spurious correlations that hinder reliable assessment. To address this, we propose a causality-inspired representation learning framework with spatiotemporal memory for polyp detection in colonoscopy videos (CIRL-Polyp). Specifically, we develop a novel View-Shift-Aware Causal Intervention Module (VACIM) to remove non-causal influences by enforcing prediction invariance under view shift perturbations. To further constrain non-causal factors, we introduce a dual-branch detection framework that processes the original and intervention frame sequences in parallel and enforces prediction consistency across branches, thereby promoting invariance to non-causal variations. In addition, we propose Causal Temporal Consistency Memory (CTCM) to leverage long sequence-dependency features and stabilize causal representations by constructing memory banks across branches and performing temporal consistency-enhanced cross-attention. Comprehensive experiments on two public video datasets and a private dataset demonstrate that CIRL-Polyp outperforms existing methods, which validates the effectiveness and suggests the clinical potential of the proposed framework from a causal perspective.

Colorectal cancer (CRC) exhibits significant heterogeneity in response to immunotherapy that cannot be fully explained by microsatellite status alone. Although elevated bile acid levels are recognized as an important risk factor for CRC, their impact on immunotherapy responsiveness remains poorly understood. Here, we demonstrate that high bile acid levels profoundly impair anti-PD-1 efficacy in both CRC patient cohort and mouse models, accompanied by reduced infiltration and functional impairment of tumor-infiltrating CD8+ T cells. Bile acid profiling identified deoxycholic acid (DCA) as the key bile acid species mediating this immunosuppressive effect. In vitro and in vivo studies have shown that DCA not only suppressed CD8+ T cell effector function but also drove them toward terminal exhaustion, thereby limiting responsiveness to anti-PD-1. Mechanistically, DCA disrupted mitochondrial fitness in CD8+ T cells by suppressing oxidative phosphorylation and inducing excessive mitochondrial reactive oxygen species (mtROS) production. In parallel, DCA enhanced ubiquitination-dependent degradation of Parkin, thereby inhibiting mitophagy and causing the accumulation of damaged mitochondria. These convergent defects in mitochondrial homeostasis ultimately promoted CD8+ T cell dysfunction and terminal exhaustion. Notably, pharmacological reactivation of mitophagy via Urolithin A reversed these defects and restored the antitumor efficacy of anti-PD-1 in vivo. Collectively, our findings identified a DCA-Parkin-mitophagy axis that drives CD8+ T cell terminal exhaustion and compromises immunotherapy efficacy, providing a potential metabolic intervention strategy to improve immunotherapy responses in CRC patients with elevated bile acid levels.

Chronic inflammation resulting from unresolved tissue injury is a potent driver of tumorigenesis. Macrophages are essential for efferocytosis, the clearance of apoptotic cells, which prevents secondary necrosis and promotes inflammation resolution. However, the epigenetic mechanisms regulating this process under inflammatory stress remain incompletely understood. Here, we identify the m6A methyltransferase METTL3 as a critical regulator of macrophage efferocytosis. Using various models of tissue injury, METTL3 expression was significantly downregulated in macrophages under inflammatory conditions and was associated with defective efferocytosis, the accumulation of apoptotic cells, and exacerbated chronic inflammation. Mechanistically, we reveal that METTL3-mediated m6A modification promotes IGF2BP2/3-dependent stabilization of MFGE8 mRNA, a process that is compromised upon METTL3 deficiency. In a colitis-associated cancer model, the efferocytosis defect associated with Mettl3 deficiency exacerbated chronic inflammatory tissue injury and increased susceptibility to colitis-associated cancer. Ultimately, external administration of recombinant MFGE8 protein effectively rescued the efferocytosis defect, mitigated inflammation, and reduced tumor burden in the AOM/DSS model. Collectively, our findings identify the METTL3-m6A-IGF2BP2/3-MFGE8 pathway as an important regulator of macrophage efferocytosis that may contribute to inflammation-associated carcinogenesis. These results suggest that targeting the METTL3-MFGE8 axis may represent a strategy for restoring efferocytosis and promoting inflammation resolution in chronic inflammatory diseases and inflammation-associated malignancies.

Nerves and cancer-associated fibroblasts (CAFs) have each been shown to regulate cancer progression directly. However, whether these cells interact to control tumor progression remains largely unknown. We show that in colorectal cancer (CRC), cholinergic stimulation induces CHRM3/Gq-dependent NTN1 secretion from CAFs, which in turn enhances intratumor cholinergic innervation. Within this feedforward loop, cholinergic stimulation promotes CRC growth directly through tumoral CHRM3/Gq-mediated YAP activation, while CAF-derived NTN1 promotes CRC growth and epithelial-to-mesenchymal transition-like programs via UNC5B-PI3K/AKT signaling. Chemogenetic activation of cholinergic neurons or fibroblast activation of the M3 receptor/Gq promotes tumoral YAP and AKT signaling and CRC progression. Conversely, blocking CHRM3 or NTN1 suppresses these pathways and improves mouse survival. In human CRC, high NTN1 expression is associated with a mesenchymal-like subtype and poor patient outcomes. These findings suggest that the neuro-mesenchymal interaction is central to CRC progression and could be therapeutically targeted with a CHRM3 antagonist or NTN1-blocking antibody.

Therapy resistance is attributed to over 80% of cancer deaths per year, emphasizing the urgent need to overcome this challenge for improved patient outcomes. Despite its widespread use in colorectal cancer (CRC) treatment, resistance to 5-fluorouracil (5FU) remains poorly understood. As an antimetabolite, 5FU imposes substantial metabolic stress, forcing cells that survive treatment to rapidly adapt. We explored acute 5FU-driven changes in mitochondria, the organelle critical for coordinating metabolic stress responses. Here we demonstrate in a range of CRC models that 5FU treatment promotes mitochondrial biogenesis and increases mitochondrial function in surviving cells. Furthermore, we show that targeting mitochondrial metabolism, particularly by inhibiting Complex I, sensitizes CRC cells to 5FU, resulting in delayed tumour growth and prolonged survival in preclinical models. Additionally, analysis of patient data suggests that oxidative metabolism signatures may predict responses to 5FU-based chemotherapy. These findings shed light on mechanisms underlying 5FU resistance and propose a rational strategy for combination therapy in CRC, emphasizing the potential clinical benefit of targeting mitochondrial metabolism to overcome resistance and enhance patient outcomes.

Perineural invasion (PNI) is a common pathological feature associated with poor prognosis of colorectal cancer (CRC). A better understanding of the mechanisms underlying PNI formation could help identify potential strategies to inhibit tumor progression. Here, we revealed an integral role of CD4+ T cells in the development and progression of PNI in CRC. Single-cell RNA sequencing, spatial transcriptomics, and multiplex immunohistochemistry profiling uncovered a distinct HSPA6+CD4+ T cell subset expressing T stress (Tstr) cell markers in CRC PNI tissues. Schwann cells (SCs) colocalized with the HSPA6+CD4+ Tstr cells in PNI, and functional studies showed that SCs promoted Tstr cell differentiation while Tstr cells enhanced SC migration. Interaction between SCs and CD4+ T cells activated the MAPK pathway in SCs and the JAK-STAT pathway in Tstr cells, and inhibition of MAPK and STAT signaling suppressed nerve invasion of CRC cells in vivo. Moreover, SCs in contact with tumor cells possessed the ability to recruit CD4+ T cells; tumor-derived TNF-α stimulated SCs to secrete CCL4, thereby recruiting CD4+ T cells toward SCs. The anti-TNF-α monoclonal antibody infliximab reversed the pro-invasive and pro-migratory effects of SCs on CRC cells and reduced intratumoral CD4+ Tstr cells, and the combination of infliximab with anti-PD-1 treatment produced a combinatorial tumor suppressive effect. Together, this study suggests that SCs near CRC tissues recruit CD4+ T cells and promote their conversion to CD4+ Tstr cells to establish an immunosuppressive microenvironment and simultaneously enhance the migration and tumor-promoting capabilities of SCs, thereby accelerating PNI development.

Improved methods to identify therapeutically relevant tumor neoantigens and their cognate T cells would aid the development of precision medicines for cancer. Here, we developed Slide-GoTags, a droplet-based single-nucleus spatial transcriptomics approach that characterizes neoantigen-specific immunity by integrating targeted transcript genotyping and T cell receptor (TCR) sequencing with single-nucleus RNA sequencing from the same slice of frozen tissue. Application of Slide-GoTags to mouse and human tumors revealed colocalization of clonally expanded, neoantigen-specific T cells with tumor cells expressing their cognate neoantigen. We also identified distinct spatial immune landscapes shaped by anti-PD1 or anti-CTLA4 blockade in mouse colorectal tumors. Across human tumor types, Slide-GoTags detected TCR-neoantigen interactions through spatial proximity and identified an enrichment of interferon-driven immunogenicity niches in immunologically 'hot' tumors compared to 'cold' tumors. These niches harbored three T cell clonotypes that colocalized with genotyped neoantigens, highlighting a spatially organized antitumor immune response. Collectively, Slide-GoTags establishes a framework for in situ mapping of T cell-tumor interactions directly from individual tissue.

Colorectal cancer (CRC) ranks as the third most common malignancy worldwide, with metastasis representing the primary cause of mortality. Aberrant activation of the Wnt/β-catenin pathway drives epithelial‒mesenchymal transition (EMT) and CRC metastasis, making β-catenin a key therapeutic target. Josephin domain containing 2 (JOSD2), a deubiquitinase with protumorigenic roles in multiple cancers, has an undefined function in CRC metastasis. Herein, by screening a protein homeostasis-related inhibitor library, we identified HY041004, a reported JOSD2 inhibitor, which potently suppresses β-catenin transcriptional activity. Mechanistically, JOSD2 modulates β-catenin protein abundance and functional activity via the RAS-ERK signaling cascade rather than through direct deubiquitination of β-catenin. Functional assays further demonstrated that JOSD2 inhibition via RNA interference or pharmacological inhibition significantly attenuated CRC metastasis both in vitro and in vivo. Collectively, our findings identify JOSD2 as a critical oncogenic factor that promotes β-catenin activity and validate JOSD2 as an underlying therapeutic target for metastatic CRC (mCRC).

Oral chemotherapy for colorectal cancer (CRC) is limited by poor tumor selectivity and microenvironment-driven resistance. Addressing these limitations demands materials that integrate tumor-selective targeting with immune microenvironment modulation. Here, clinical analysis of CRC specimens revealed pronounced transferrin (Tf) enrichment in CRC-associated intestinal regions. Guided by this finding, we engineered a gastrointestinal-stable cyclic Tf-binding peptide (cp) with high Tf affinity and constructed cp-modified ginger-derived extracellular vesicles (cp-GEVs) for in situ Tf-mediated sandwich-like targeting. By recruiting endogenous Tf, cp-GEVs established a Tf-mediated bridging interface that selectively engages Tf receptor-overexpressing intestinal epithelium and tumor cells, enabling efficient epithelial transcytosis, tumor-selective accumulation, and deep intratumoral penetration after oral administration. When loaded with irinotecan (CPT-11), CPT@cp-GEVs significantly enhanced intracellular drug delivery and reprogrammed immunosuppressive M2-like tumor-associated macrophages toward a pro-inflammatory phenotype, thereby disrupting cancer stem cell-enriched drug-resistant niches. In AOM/DSS-induced primary CRC models and patient-derived ex vivo systems, CPT@cp-GEVs significantly improved chemotherapeutic efficacy while attenuating resistance. Collectively, this work establishes a Tf-mediated sandwich-like targeting framework for oral cancer therapy, offering a conceptually distinct materials design paradigm that integrates endogenous ligand recruitment with immune microenvironment reprogramming.

Microsatellite instability-high (MSI-H) colorectal cancer cells depend on the Werner syndrome helicase (WRN) to resolve cruciform DNA structures that arise from expanded TA-dinucleotide repeats. Loss of WRN induces replication stress and double-strand breaks (DSBs), a vulnerability that can be recapitulated by the selective WRN inhibitor HRO761 in MSI cancer cells. To uncover the mechanisms governing sensitivity to WRN inhibition, we conducted genome-wide CRISPR/Cas9 screens in colorectal cancer cell lines treated with or without HRO761. These screens identified SMARCAL1 as a key modulator of WRN dependency. Depletion of SMARCAL1 rendered cells resistant to WRN inhibition, and rescue of this effect required the ATPase/translocase activity of SMARCAL1. Mechanistically, SMARCAL1 antagonized WRN and supported cruciform DNA structures, thereby enhancing cellular reliance on WRN. In addition, the MRE11-RAD50-NBS1 (MRN) complex, rather than MUS81 or ERCC1/XPF, was the principal mediator of cruciform DNA processing following WRN inhibition. Acute disruption of the MRN complex conferred profound resistance to WRN inhibition, whereas ATM deficiency produced a more modest resistant phenotype. Further genetic and pharmacological epistasis analyses demonstrated that the MRN complex regulates WRN inhibitor sensitivity through both ATM-dependent signaling and MRE11 nuclease-dependent functions. Importantly, the key resistance mechanisms identified in this study were independently validated using a structurally distinct clinical-stage WRN inhibitor VVD-214. Collectively, these findings identify the SMARCAL1-MRN-ATM axis as a critical regulator of WRN dependency and provide mechanistic insight into resistance to WRN-targeted therapy.

The peritumoral microenvironment has emerged as a key role in affecting tumor invasion and immunotherapy responses. In adipose-enriched tumors, such as breast cancer (BC), peritumoral adipose tissue (PA) harbors unconventional immune populations, yet its immunological functions remain poorly understood. In particular, how adipocyte regulate innate-like lymphocytes, such as γδ T cells, remains unclear. We performed single-cell RNA sequencing and spatial profiling of paired specimens from patients with BC. Integrated multi-omics analyses, immunofluorescence staining, human γδ T-cell expansion assays, functional assays, and in vivo models were used to define the immune cell states and evaluate the impact of lipid mediator leukotriene B4 (LTB4) on γδ T-cell activation and signaling. Clinical correlations were assessed using our cohort and patient datasets. We identified a previously unrecognized population of NKG2A+γδ T cells with predominant Vδ2 usage that preferentially accumulated in PA, particularly at the adipose-tumor interface. Multi-omics analyses revealed that they exhibited potent cytotoxic activity and extensive interactions with dendritic cells, coordinating a local immune surveillance network. Mechanistically, peritumoral adipocytes showed enhanced activation of the 5-lipoxygenase pathway and secreted the lipid mediator LTB4, which selectively combined to LTB4 receptors, thereby activating STAT1 signaling in γδ T cells and upregulating NKG2A expression. NKG2A+γδ T cells were preferentially enriched in ductal carcinoma in situ and early-stage BC. These cells were also associated with favorable clinical outcomes across multiple adipose-enriched tumors. Assays using human specimens confirmed that LTB4 potentiated γδ T cell-mediated antitumor responses. Importantly, LTB4-programmed γδ T cells displayed superior killing capacity using in vitro and in vivo assays. These findings redefine PA as an active immunological niche that programs γδ T-cell immunity through adipocyte-derived LTB4 signaling. Our study identifies NKG2A+γδ T cells as cytotoxic sentinels at the adipose-tumor interface during early tumor development, providing a rationale for leveraging LTB4-mediated γδ T-cell programming in translational cancer immunotherapy.

Hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2-) breast cancer accounts for approximately 70% of breast cancer cases. Despite recent advances with cyclin-dependent kinase 4/6 inhibitors (CDK4/6i), resistance inevitably develops, often driven by activation of the phosphatidylinositol 3-kinase (PI3K)-AKT-mammalian target of rapamycin (mTOR) pathway. Genetic alterations such as PIK3CA mutations (present in ~ 45% of HR+/HER2- tumors), AKT1 mutations, and PTEN loss contribute to endocrine resistance and poor outcomes. This review summarizes emerging strategies targeting this pathway to overcome resistance in advanced disease. Isoform-specific PI3K inhibitors, including alpelisib and inavolisib, have demonstrated clinically meaningful progression-free survival benefits in PIK3CA-mutated populations, with inavolisib showing improved tolerability and efficacy. In contrast, pan-PI3K inhibitors such as buparlisib have been constrained by toxicity. Targeting downstream signaling, AKT inhibitors have also shown benefit: capivasertib has demonstrated clinical efficacy leading to US Food and Drug Administration approval, while ipatasertib has yielded encouraging results, particularly in tumors harboring PIK3CA, AKT1, or PTEN alterations. Mammalian target of rapamycin inhibitors, notably everolimus, have shown efficacy irrespective of mutation status. The dual PI3K-mTOR inhibitor (gedatolisib) has also shown promising progression-free survival benefit in a PIK3CA wild-type population. Next-generation agents, including mutant-selective PI3Kα inhibitors and bi-steric mTOR complex 1 inhibitors, are under active investigation. Optimal sequencing of these agents alongside endocrine therapy and CDK4/6i options remain a critical question, as does integration of genomic testing to guide therapy. Future directions include rational combination strategies, improved biomarker-driven selection, and novel modalities such as proteolysis-targeting chimeras (PROTACs). Collectively, these advances aim to enhance durability of response, minimize toxicity, and improve survival in HR+/HER2- metastatic breast cancer.

Neoadjuvant therapy (NAT) has emerged as a standard treatment strategy for locally advanced breast cancer (BC), yet robust biomarkers for response prediction remain elusive. Here, we established a real-world NAT cohort of 1161 Chinese BC patients, including 1145 cases with matched clinicopathological data and targeted sequencing, to systematically evaluate genomic features associated with NAT outcomes. We identified both cross-subtype and subtype-specific genomic associations with treatment response. PI3K-pathway alterations emerged as a consistent feature of resistance across subtypes, whereas mutations such as ERBB2 in HER2+ disease and MAP3K1 in triple-negative breast cancer were associated with subtype-specific response patterns. Regimen-level analyses further showed that some genomic associations were treatment-context dependent across chemotherapy-, endocrine-, anti-HER2-, and immunotherapy-containing regimens. Among patients with non-pathological complete response (non-pCR), genomic profiling further refined risk stratification for distant recurrence by revealing subtype-specific prognostic alterations, including TOP3B and SETD2. Furthermore, a machine-learning model integrating genomic and clinicopathological features showed favorable performance for NAT response prediction. Overall, our study provides a comprehensive genomic framework for response prediction and recurrence risk assessment, supporting more precise stratification and biomarker-guided treatment optimization in Asian breast cancer patients.

In TROPION-Breast01 (NCT05104866), datopotamab deruxtecan (Dato-DXd) improved progression-free survival by blinded independent central review versus investigator's choice of chemotherapy (ICC) in patients with inoperable/metastatic hormone receptor-positive, human epidermal growth factor receptor 2-negative (HR+/HER2-) breast cancer, who had disease progression on endocrine therapy and for whom endocrine therapy was unsuitable, and who had received 1-2 prior lines of chemotherapy in the inoperable/metastatic setting. We report detailed safety data and patient-reported outcomes (PROs) from the final analysis. Patients received Dato-DXd (6 mg/kg every 3 weeks) or ICC (eribulin/capecitabine/vinorelbine/gemcitabine). Mouthwash was advised (steroid mouthwash strongly recommended for Dato-DXd), and ophthalmologic assessments were carried out per protocol. Safety and time to deterioration (TTD) in global health status/quality of life (GHS/QoL), pain, and physical functioning, using the European Organisation for Research and Treatment of Cancer QoL Questionnaire-Core 30, were secondary endpoints; exploratory PROs were also assessed. With Dato-DXd (n = 360), nausea was the most common treatment-related adverse event (AE; 51.9%, grade ≥3: 1.4%). The AEs of special interest (grouped terms), treatment-related oral mucositis/stomatitis and ocular surface events, occurred in 57.2% (grade ≥3: 7.2%) and 43.9% (grade ≥3: 1.9%) of patients, respectively. Per protocol recommendations, most patients receiving Dato-DXd received anti-emetics (76.7%), prophylactic mouthwash (77.2%), or artificial tears (71.7%). Adjudicated drug-related interstitial lung disease/pneumonitis with Dato-DXd was mostly low-grade (3.9%; grade ≥3: 0.8%; one grade 5 event). Hematologic toxicity was the most notable feature of the ICC safety profile (n = 351). TTD in GHS/QoL, pain, and physical functioning was delayed with Dato-DXd versus ICC. Patient-reported symptomatic AEs were generally consistent with the clinician-reported AE profile in both arms. Safety and PRO data from TROPION-Breast01 complement the improvements in efficacy demonstrated with Dato-DXd versus ICC, supporting Dato-DXd as a treatment option for patients with previously treated, inoperable/metastatic HR+/HER2- breast cancer.

Comprehensive genomic profiling increasingly identifies BRCA alterations in breast cancer (BC). While germline BRCA mutations are well established as biologically and clinically relevant, the significance of somatic BRCA alterations remains unclear. We hypothesized that the implications of BRCA alterations may differ according to germline BRCA alterations. Among 5411 BC cases registered in the Center for Cancer Genomics and Advanced Therapeutics database, we analyzed 2978 BC cases with available germline (g) and somatic (s) BRCA status. Patients were categorized into four groups (g+/s+, g+/s-, g-/s+, and g-/s-). Mutational landscapes, homologous recombination deficiency (HRD)-related alterations, specimen origin, and time to treatment discontinuation (TTD) were evaluated, focusing on estrogen receptor (ER)-positive/HER2-negative and triple-negative breast cancer (TNBC). g-/s+ tumors were more prevalent than g+/s+ tumors, and concordance between germline and somatic BRCA status was limited. Somatic BRCA alterations were more frequently detected in metastatic lesions and were more frequently observed in metastatic specimens. Genomically, g-/s+ tumors were enriched for oncogenic signaling mutations, including PIK3CA and TP53, whereas g+/s+ tumors showed HRD-related features dominated by BRCA alterations. Non-BRCA HRD-related alterations were most frequent in g-/s+ tumors (52%; P = 0.004). In ER-positive/HER2-negative BC, germline BRCA positivity was associated with shorter cyclin-dependent kinase 4 and 6 inhibitor TTD, and g+/s+ status remained independently associated with shorter TTD (hazard ratio 1.3, P < 0.001), whereas g-/s+ tumors were comparable with g-/s- tumors. In contrast, g-/s+ tumors showed TTD comparable with g-/s- tumors. In TNBC, immune checkpoint inhibitor outcomes were not associated with BRCA status. BRCA alteration groups defined by combined germline and somatic status showed distinct genomic and clinical patterns in BC. Somatic BRCA alterations should not be interpreted in isolation and require integration with germline status and broader genomic context.

Breast cancer (BC) is a highly complex and heterogeneous malignancy and the most prevalent cancer among women worldwide. The diagnosis, prognosis, and the treatment of BC pose significant challenges that are responsible for their limited therapeutic efficacy. Omics-based technologies have gained substantial attention in BC diagnosis through molecular profiling and diverse clinical analytics. The integration of metabolomics, proteomics, transcriptomics, and genomics provides a multidimensional approach to personalized BC diagnosis and treatment through high-throughput molecular profiling. Moreover, the emergence of artificial intelligence (AI) has also supported more accurate and early diagnosis of BC through multimodal integration of diverse datasets. The integration of advanced deep learning (DL) and machine learning (ML) has been extensively exploited for tumor grading, histopathological classification, molecular profiling, diagnostic imaging, and prognostic prediction. This review aims to summarize recent developments in AI-driven multi-omics approaches for the discovery of BC biomarkers. We have also highlighted the integration of omics-based data like metabolomics, proteomics, transcriptomics, and genomics with key AI techniques, including ML and DL, that play a crucial role in the inclusion of multi-omics in cancer and biomarker discovery. We have further discussed AI-based BC screening and diagnostic approaches, as well as the contribution of AI models for patient stratification, biomarker discovery, and prediction of therapeutic response. Additionally, key limitations and challenges, including data heterogeneity, high computational complexity, and model interpretability, have also been highlighted in the present review. Conclusively, we have also outlined future perspectives on the integration of AI and multi-omics to revolutionize precision clinical medicine and improve clinical outcomes in BC theranostics.

Currently, three cyclin-dependent kinase 4 and 6 inhibitors (CDK4/6i) are approved in combination with endocrine therapy (ET) as first-line treatment of patients with hormone receptor (HR)-positive/human epidermal growth factor receptor 2 (HER2)-negative metastatic breast cancer (MBC). The impact of homologous recombination repair (HRR) pathogenic variants (PV) on outcomes with first-line CDK4/6i plus ET in HR-positive/HER2-negative MBC remains uncertain. We conducted a multicenter, real-world, case-control study including 233 patients with HR-positive/HER2-negative MBC treated with first-line CDK4/6i and ET. Among them, 116 presented HRR PVs and 117 were matched controls with negative germline testing. The primary objective was to compare progression-free survival (PFS) and overall survival among germline-BRCA2 PV carriers, other HRR PV carriers, and controls. To minimize baseline differences in prognostic factors between PV carriers and controls, inverse probability of treatment weighting was applied. Molecular analyses in pre-CDK4/6i samples among patients with BRCA2 PV were carried out, including RAD51-foci, PAM50 intrinsic subtype, and RB1 loss of heterozygosity (LOH). Among the included 233 patients, median age at diagnosis was 45 years (interquartile range 39-56) and 33% had de novo metastatic disease. Primary resistance to adjuvant ET was present in 10% and secondary resistance in 27%. After a median follow-up of 44 months, patients with germline-BRCA2 PVs (n = 67) had significantly shorter PFS [11 versus 27 months; adjusted hazard ratio (aHR) 2.73, 95% confidence interval (CI) 1.65-4.51, P < 0.001] compared with controls. Among patients with endocrine-sensitive disease, germline BRCA2 PV carriers had markedly shorter PFS (median PFS 12 versus 39 months; aHR 4.04; 95% CI 1.82-8.98, P < 0.001). Exploratory analyses revealed RB1 LOH before CDK4/6i-treatment in most evaluable BRCA2 tumors. BRCA2 PVs were independently associated with poorer outcomes to first-line CDK4/6i plus ET in HR-positive/HER2-negative MBC compared with controls, especially relevant among patients with endocrine-sensitive disease. These findings suggest that patients with a germline PV in BRCA2 may require alternative first-line strategies.

Second near-infrared (NIR-II, 1000-1700 nm) fluorescence imaging (FLI)-guided photothermal therapy is a promising non-invasive strategy for tumor theranostics. However, existing photothermal agents struggle to balance high photothermal conversion efficiency (PCE) with strong fluorescence quantum yield (QY). Herein, an alkoxyl side chain engineering strategy was proposed to construct a series of donor-π bridge-acceptor-π bridge-donor (D-π-A-π-D) structured organic photothermal agents (TTQ1-TTQ6) enabling simultaneous NIR-II FLI-guided photothermal therapy. By introducing different alkoxyl side chains on triphenylamine donor or thiophene π-bridge, the photophysical properties including aggregation-induced emission (AIE) property, QY and PCE were regulated. TTQ2 NPs and TTQ6 NPs featured superior resolution and low background in in vivo NIR-II fluorescence imaging of the mouse blood vessels with high signal-to-noise ratio. Four alkoxyl chains attached to the donor moiety enhanced the intramolecular donor-acceptor interaction and effectively suppressed aggregation-caused quenching (ACQ), enabling TTQ6 NPs to exhibit desirable NIR-II fluorescence and a high PCE of 46.5%. Encouragingly, TTQ6 NPs could effectively ablate both subcutaneous and orthotopic breast tumors and stimulate adaptive immune responses, which collectively reduced the risk of tumor recurrence. This study illustrates the potential of alkoxyl side chain engineering strategy in developing advanced photothermal agents for NIR-II FLI-guided theranostics.

Surgical resection remains the primary treatment in human breast cancer. However, the accurate and rapid delineation of tumor margins intraoperatively is still a significant challenge. Herein, we designed a near-infrared fluorescent probe DCIP-ENTK by incorporating dicyanoisophorone dye with a negatively charged peptide, which can be specifically cleaved by enterokinase (ENTK, a tumor biomarker for breast cancer). In vitro experiments confirmed that the probe could quickly (<10 min), specifically, and sensitively (LOD = 0.031 U/mL) respond to ENTK. Notably, this probe was engineered as a peptide-based architecture that can self-assemble into a nanoprobe. This design not only provides excellent biocompatibility and in vivo stability, but also enables specific mitochondria-targeting within cancer cells. Upon cleavage by ENTK, the released hydrophobic fluorophores aggregated at the enzyme-rich site. This unique aggregation-induced retention effect enabled long-term, high-contrast in situ imaging of tumor margins in tumor-bearing mice and human breast cancer samples, which offers a powerful and clinically translatable tool for improving the precision of cancer surgery. Finally, we also achieved the quantitation of ENTK levels in artificial human duodenal juice with recovery rates from 93.5% to 99.8%. These results confirm the suitability of our probe for the quantitation of ENTK in real samples.

Chimeric antigen receptor (CAR) macrophage therapy shows significant potential for solid tumors owing to the intrinsic tumor infiltration and phagocytic capacity of macrophages. However, its clinical translation is limited by macrophage phenotypic plasticity within the immunosuppressive tumor microenvironment and the complexity of ex vivo cell manufacturing. It is essential to develop techniques that enable macrophages to be activated specifically by antigens while sustaining their proinflammatory activity in vivo. Here, we report a mannose-modified lipid nanoparticle (LNP) platform for the co-delivery of CAR-encoding messenger RNA (mRNA) and the Toll-like receptor (TLR) 7/8 agonist resiquimod (R848), enabling in situ generation of proinflammatory CAR macrophages. In vitro, we assessed macrophage-preferential uptake, CAR expression efficiency, TLR7/8 agonist-mediated macrophage polarization, and immune activation. In vivo efficacy was assessed in syngeneic and humanized mouse models of triple-negative breast cancer, including postoperative recurrence and lung metastasis models. Systemic administration of M-LNP/CAR+R848 induced robust CAR expression in tumor-associated macrophages and promoted sustained M1 polarization. Engineered macrophages exhibited enhanced antigen-specific phagocytic activity and tumor cell clearance, and promoted CD8+ T cell proliferation and NK cell infiltration, thus coordinating innate and adaptive immune responses. Functional macrophage depletion experiments demonstrated that tumor control was dependent on macrophages. In vivo treatment significantly reduced the growth of primary tumors, prevented postoperative recurrence, and prolonged survival in mice with lung metastases in both syngeneic and humanized models. Our findings demonstrate that M-LNPs enabling co-delivery of mRNA and an innate immune agonist enable in situ generation of proinflammatory CAR macrophages and induce durable antitumor immunity. This controllable and non-integrative strategy allows tunable immune activation, provides a flexible platform for CAR macrophage-based immunotherapy in triple-negative breast cancer.

Adipocytes are essential stromal components of the tumor microenvironment (TME) in breast cancer that play pivotal roles in cancer progression and chemoresistance. In close proximity to tumor cells, they undergo phenotypic reprogramming into cancer-associated adipocytes (CAAs), characterized by multilocular lipid droplets, increased mitochondrial content, and elevated expression of uncoupling protein 1 (UCP1). Although these features superficially resemble those of beige adipocytes, they do not recapitulate classical thermogenic programming, reflecting a unique metabolic adaptation driven by the TME. Here, we identified tumor necrosis factor receptor-associated protein 1 (TRAP1), a mitochondrial paralog of HSP90, as a central regulator of the transition of adipocytes into CAAs. TRAP1 was highly upregulated in CAAs and was required to drive a tumor-associated adipocyte secretory program, including the adipokine complement factor D (CFD). Genetic and pharmacological TRAP1 inhibition destabilized the mitochondrial electron transport chain, reduced cellular respiration, and activated the energy sensor AMPK. This subsequently suppressed mTOR and PPARγ signaling, effectively abrogating adipocyte reprogramming and diminishing pro-tumorigenic adipokine secretion. Crucially, this CAA-secreted CFD promoted cancer cell survival and chemoresistance via C3aR-AKT/ERK signaling, and blocking this TRAP1-mediated crosstalk profoundly sensitized breast tumors to chemotherapy in vivo. Collectively, these findings identify TRAP1 as a master regulator of adipocyte transdifferentiation within the TME, offering a novel strategy to restrict tumor growth and overcome drug resistance in breast cancer.

Lack of DNA double-strand break repair efficiency exquisitely sensitizes cancers to poly-ADP ribose polymerase inhibitors (PARPi). Unfortunately, resistance to PARPi poses an insurmountable challenge for patients. Mechanisms that confer insensitivity to PARPi therapy include enhanced DNA damage repair and autophagy. Natural and non-natural unsaturated fatty acid nitroalkene derivatives (NFA) show anticancer actions that sensitize TNBC cells to PARPi and other DNA-damaging treatments. We reveal that nitro-oleic acid (OA-NO2) re-sensitizes PARPi-resistant TNBC cells to PARPi. RNA-seq analysis of clinically relevant mutBRCA1 PARPi-resistant TNBC cell lines exhibited upregulation in autophagy and lysosomal pathways. Bio-orthogonal analysis identified the autophagy regulator SQSTM1/p62 as a novel OA-NO2 target, alkylating two redox-sensitive Cys residues of p62 (Cys105 and Cys113). These Cys are essential for p62 regulation of autophagy and mimicked the effects of p62 Cys105 and Cys113Ala mutants and when alkylated by OA-NO2 showed impaired p62 oligomerization, degradation, and inhibition of autophagy. Combination treatment of PARPi-resistant TNBC with a PARPi and OA-NO2 identified the most synergistic HSA scores and inhibited p62-associated autophagy and lysosome function. These data underscore the clinical potential of OA-NO2 for treating PARPi-resistant TNBC patients.

Estrogen receptor-positive (ER+) breast cancer is the most common subtype of breast cancer and is an age-related disease. How systemic and tumor microenvironment hormone signaling intersects with chronic inflammation and how this shapes aging-associated tumor biology remain incompletely understood. Here, using an aged rat model of ER+ tumors, we identify age-associated differences in tumor development and immune features. In parallel, analysis of samples from patients with ER+/HER2- breast cancer and age-matched controls demonstrated estrone-predominant systemic estrogen patterns and increased tumor HSD17B7 expression in older patients, consistent with enhanced local estrone-to-estradiol conversion. In patient-derived organoids from older women, pharmacologic inhibition of HSD17B7 reduced estrogen conversion and proliferation-associated transcriptional programs. Tumors from older patients also exhibited chemokine enrichment, including CCL2, associated with immunosuppressive macrophage features. Targeting both estrogen signaling and chemokine pathways attenuated macrophage polarization ex vivo. These findings support a model linking age-associated hormonal and inflammatory changes to features of a tumor-permissive microenvironment in ER+ breast cancer.

Cancer stem cells (CSCs) drive metastasis and therapy resistance, yet their behaviour within the complex tumour microenvironment remains poorly understood. Here we use a fluorescent reporter that marks CSCs to show that CSCs and their more differentiated progeny display strikingly different population dynamics during metastatic lung colonization in breast cancer models. CSC expansion is rapidly curtailed early in colonization, suggesting a strong negative feedback mechanism acting selectively on this subpopulation. We showed that CSCs are exceptionally sensitive to local microenvironmental cues such as cell crowding and nutrient availability. They respond earlier and more extensively than their differentiated progeny, thereby coupling tumour growth to resource and space availability. Microenvironmental signals converge on the transcriptional regulatory complex YAP/TAZ/TEAD, with CSC sensitivity arising from elevated signal reception and greater chromatin accessibility at TEAD-regulated enhancers. Targeting upstream inputs to this pathway reversed chemotherapy-induced CSC enrichment in lung metastases, suggesting a potential therapeutic strategy.

Heat shock protein 90 (Hsp90) is an established and clinically validated anticancer target. By regulating the folding and maturation of diverse client proteins, it is indirectly involved in carcinogenesis. Key breast cancer oncogenic drivers, including estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2, are established Hsp90 clients, making breast cancer models widely used for Hsp90 inhibitor evaluation. However, N-terminal inhibitors are well known to induce a heat shock response (HSR), representing a major limitation in cancer therapy. This has driven the development of alternative modalities, such as C-terminal inhibitors, which in several cases avoid HSR induction. To evaluate Hsp90 inhibitors and characterize their proteomic effects, most studies employ western blotting (WB) as the primary method. On WB, different inhibitors are tested across various concentrations, time points, and breast cancer cell lines for their effects on client, heat shock, and other proteins. This generates extensive data sets that outline the expected WB performance of Hsp90 inhibitors. In this review, we focus on 10 protein families that are commonly used to assess Hsp90 inhibition in in vitro breast cancer models and represent all 10 cancer hallmarks. We provide basic structural and functional information on these families and comprehensively describe the effects of N- and C-terminal inhibitors on their levels. We summarize WB-based observations for more than 100 Hsp90 inhibitors, outlining the characteristic effects of N- and C-terminal inhibition, highlighting opportunities for future research, and ending with a suggestion for the WB method design based on best practices observed in literature.

In breast cancer, local invasion of cancer cells into surrounding tissue marks the first step of metastasis. However, to elucidate the impact of cells from the tumor microenvironment on this process, advanced 3D migration models are still urgently needed. To enable migration and invasion studies in a fully 3D bioprinted tumor-stroma model, a migration-permissive bioink composed of methacrylated collagen type I and thiolated hyaluronic acid with low polymer content is developed. In a printed co-culture model comprising metastatic breast cancer cells (MDA-MB-231) and adipose-derived stromal cells (ASCs), real-time single-cell tracking reveals that ASCs in the stromal compartment profoundly promote migration and invasion dynamics of individual tumor cells. This is reflected by increased speed, migration distance, and invasion into the stroma, and is accompanied by collagen remodeling and a shift in tumor cell morphology. A correlation between tumor cell morphology and migration speed is evident, which is modulated by ASCs. A highly motile and invasive subset of tumor cells is significantly enhanced in the presence of ASCs. These insights into the influence of ASCs on the heterogeneity of breast cancer cells in terms of their migratory behavior may inform the development of more specific and effective treatment options for metastatic breast cancer.

The design of organic photosensitizers that operate efficiently in the near-infrared (NIR) region remains challenging due to the need to balance electronic delocalization, stability, and biological compatibility. Here we report a rational, electronegativity-guided strategy for engineering (donor-π)3-acceptor pyrylium-based chromophores capable of concurrent NIR phototherapy and subcellular imaging. Density functional theory calculations identified pyrylium (PYR) as the optimal acceptor for narrowing the HOMO-LUMO gap and enhancing intramolecular charge transfer (ICT). The resulting octupolar chromophores, PYR-OMe and PYR-NPh2, display intense absorption in the 600-800 nm range, large Stokes shifts (>100 nm), and excellent photostability. Under NIR irradiation, PYR-NPh2 catalyzes efficient NADH photooxidation and generates multiple reactive oxygen species (ROS), even under hypoxic conditions. In cancer cells (e.g., MDA-MB-231), PYR-NPh2 induces redox imbalance and pronounced phototoxicity while enabling high-fidelity lysosomal imaging at ∼100 nm resolution using structured illumination microscopy. This work introduces a unified molecular design principle for NIR organic chromophores that integrate redox modulation, ROS generation, and organelle-level visualization, advancing the frontier of precision phototheranostics.

R-loops are transcription-induced, three-stranded nucleic acid structures that, if not properly resolved, can disrupt DNA repair and compromise genome stability. BRCA2, a tumor suppressor vital for homologous recombination (HR), also contributes to R-loop regulation, though the underlying mechanisms remain poorly understood. Here, we identify HELZ as a direct BRCA2 interactor and characterize it as an ssRNA-specific R-loop resolvase. BRCA2 enhances HELZ helicase activity and promotes its recruitment to R-loops. Importantly, HELZ resolves R-loops at DNA double-strand breaks, enabling efficient DNA end resection and HR, particularly within transcriptionally active genomic regions. We further demonstrate that HELZ is critical for R-loop clearance in cancers with elevated transcriptional activity and R-loop accumulation, such as estrogen receptor-positive breast cancer, where it becomes essential for cell survival under estrogen-induced transcriptional stress. These findings establish HELZ as a BRCA2-dependent regulator of R-loop homeostasis and identify it as a potential biomarker and therapeutic target in R-loop-driven malignancies.

Immunosuppressive tumor microenvironment is a major driver of tumor progression and therapeutic resistance. Basal-like breast cancer (BLBC), which largely overlaps with triple-negative breast cancer, generally displays an immunosuppressive tumor microenvironment enriched with tumor-associated macrophages and other immunosuppressive cell populations. However, the driving mechanisms that establish and maintain these tumor microenvironment features remain not fully understood. This study identifies tumor cell-intrinsic POU4F1 as a key regulator of the immunosuppressive tumor microenvironment in BLBC. Tumors with high POU4F1 expression exhibited lower IFNγ-related signature and poor responses to immunotherapy. In immunocompetent 4T1 tumor-bearing mice, POU4F1 knockout led to decreased infiltration of M2-like macrophages and increased infiltration of proliferative and functional CD8+ T cells and NK cells. In-vitro assays using human immune cells demonstrated that POU4F1 directly promoted monocyte recruitment and macrophage polarization, which in turn suppressed the proliferation and effector function of tumor-specific CD8+ T cells and NK cells. Mechanistically, POU4F1 upregulated CCL2 expression through NIK-mediated activation of the non-canonical NF-κB signaling pathway, thereby promoting monocyte recruitment and immunosuppressive phenotype polarization. Genetic ablation or pharmacological targeting of POU4F1 with Bobcat339 significantly inhibited tumor growth, remodeled the tumor immune microenvironment, and synergized with anti-PD-1 therapy in mouse models and patient-derived breast cancer organoids. These findings provide mechanistic insights into how POU4F1, a BLBC-specific transcription factor, orchestrates intercellular crosstalk to establish an immunosuppressive, tumor-supportive microenvironment, and indicate that targeting POU4F1 may represent a promising therapeutic strategy for BLBCs.

This work demonstrates that the integrity of actin filaments can selectively regulate both lateral motion and in-plane orientation of transmembrane proteins on cell membranes. Using anisotropic gold nanostars functionalized with HER2 (human epidermal growth factor receptor)-targeting aptamers as optical probes, we tracked HER2 receptor dynamics on cancer cells using differential interference contrast microscopy. We found that for cells with an intact actin structure, the lateral mobility of receptors was restricted, consistent with actin-supported membrane compartmentalization. When actin filaments were disrupted after treatment with latrunculin A, HER2 receptors exhibited Brownian motion. In parallel, the receptors exhibited longer rotational dwell times corresponding to reduced orientational freedom. Analysis of receptor dynamics revealed that the coordinated translation-rotation regulation by actin filaments only occurred under directed diffusion modes when the cytoskeleton was intact. Two-dimensional single-nanoconstruct tracking provides a real-time approach to uncover how cytoskeletal architecture can organize receptor dynamics in living cells.

Lung adenosquamous carcinoma (ASC) is a rare histological subtype of non-small cell lung cancer (NSCLC), and prospective data regarding the efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) in this patient population remain scarce. This prospective, multicenter, single-arm phase II trial (NCT04354961) evaluated the safety and efficacy of aumolertinib as first-line treatment for treatment-naïve patients with stage IV EGFR-mutant ASC. Eligible patients received aumolertinib 110 mg orally once daily, with progression-free survival (PFS) as the primary endpoint and objective response rate (ORR), disease control rate (DCR), overall survival (OS), and safety as secondary endpoints. The study was terminated early due to slow accrual, with 12 patients enrolled (median age: 66 years; 58.3% male; 58.3% with EGFR exon 19 deletion). After a median follow-up of 29.0 months, the median PFS and OS were 11.1 months (95% CI, 4.27-NA) and 16.7 months (95% CI, 11.3-NA), respectively. Confirmed ORR and DCR were 58.3% and 83.3%. Treatment-related adverse events (TRAEs) occurred in 75% of patients, with 33.3% experiencing grade 3-4 TRAEs; no treatment-related deaths were observed. Aumolertinib exhibits clinical activity with a manageable safety profile in EGFR-mutant ASC patients. However, its reduced efficacy compared with EGFR-mutant adenocarcinoma highlights the need for histology-specific therapeutic strategies. Trial Registration: NCT04354961.

Stereotactic body radiation therapy (SBRT) is effective for early-stage non-small cell lung cancer (NSCLC), but the treatment of ultra-central NSCLC (UCNLC) near critical structures remains challenging. This retrospective study analyzed 66 patients with stage I-II unresectable UCNLC who underwent SBRT at Shanghai Pulmonary Hospital (2020-2023). UCNLC was defined by planning target volumes (PTVs) abutting/overlapping vital structures (proximal bronchial tree, esophagus, heart, great vessels, pulmonary vessels) or internal target volumes (ITVs) within 1 cm of these organs. Tailored SBRT plans with non-uniform PTV margins were used in 45.5% of cases, enabling 53.3% of patients to receive a biologically effective dose ≥ 100 Gy. With a median follow-up of 32.2 months, the median progression-free survival (PFS) was 42.9 months; 1- and 3-year PFS rates were 93.9% and 68.8%, respectively. Median local control (LC) and overall survival (OS) were not reached, with 1- and 3-year LC rates of 98.5% and 84.6%, and OS rates of 98.5% and 88.7%, respectively. Toxicities were predominantly grade 1-2, with only one grade 3 pneumonitis. Larger ITV predicted poorer LC and PFS, whereas T stage independently predicted PFS and OS. Therefore, a non-uniform PTV strategy maintained clinical efficacy without compromising safety, supporting its application in UCNLC management.

Despite the incorporation of atezolizumab, an anti-programmed death-ligand 1 (PD-L1) antibody, into first-line chemotherapy regimen, survival in patients with extensive-stage small cell lung cancer (ES-SCLC) remains poor. Given the role of dendritic cells (DCs) in modulating responses to anti-programmed cell death protein 1 (PD-1) antibodies, a promising avenue is the development of vaccines based on DCs, together with the evaluation of how DC phenotypic modifications shape treatment efficacy. This is a single-arm, phase Ib/II, open-label clinical trial investigating the safety and activity of combining autologous tumor lysate pulsed DCs administered intradermally with intravenous atezolizumab as maintenance therapy for patients with ES-SCLC. Following leukapheresis, monocytes were differentiated ex vivo into mature DCs and loaded with irradiated autologous tumor lysate. After standard induction therapy (four cycles of carboplatin, etoposide, and atezolizumab), patients received up to six doses of mature DCs intradermally, along with intravenous atezolizumab 1,200 mg every 3 weeks, until disease progression. 20 patients were enrolled, including six patients with brain metastases. 18 patients received DC vaccination, with a median of three doses (range 1-6). Most grade 3-4 treatment-related adverse events were hematological and related to chemotherapy. DC vaccination was well tolerated, with only grade 1-2 transient skin irritation at the injection site. Among 18 evaluable patients, 17 (94.4%) achieved an objective response. After a median follow-up of 31.4 months, progression-free survival at 2 years (PFS2y) was 20% (95% CI 6.4% to 39.1%), median overall survival (OS) 11.2 months (95% CI 6.3 to 26.0) and OS3y 22.5% (95% CI 6.8% to 43.6%). Six patients achieved long survival (median 30.5 months, range 23.4-36.2). Expansion of conventional type DC1 (cDC1s) XCR1+ and stem-like progenitor-exhausted (CXCR5+PD-1+) CD8+ T cells following chemotherapy plus atezolizumab was associated with improved PFS (HR 0.94, 95% CI 0.90 to 0.99 and HR 0.59, 95% CI 0.39 to 0.89). DC immunization combined with atezolizumab maintenance following standard chemotherapy plus atezolizumab treatment is well tolerated. Its clinical activity cannot be definitively determined in this study, as it was administered as maintenance therapy; however, the observed rate of long-term survivors suggests a potential benefit. Our findings suggest a pivotal role for cDC1s in mediating the antitumor activity in long-term survivors. NCT04487756.

Anti-angiogenic therapy benefits vary, with response rates of 40 to 70%, highlighting the need for early biomarkers to identify responders. We developed an automated machine learning framework that uses delta quantitative vascular morphometry features from standard contrast-enhanced CT to evaluate treatment response. This workflow combines automated tumor and vessel segmentation with feature extraction from routine scans for clinical use. Shapley additive explanations (SHAP)-based attributions identify key vascular and clinical features, providing meaningful, imaging-visible evidence aligned with therapy targets beyond traditional radiomics. Using baseline and follow-up CTs from 163 patients with lung cancer, we built three models using fivefold cross-validation, with the delta-merge model achieving high accuracy (area under the receiver operating characteristic curve = 0.842 internally, 0.806 externally). SHAP analysis uncovered an "arterial-dominant, venous-adaptive" pattern, where arterial involvement and venous recovery distinguish responders. This automated workflow and visualization support early, imaging-based response assessment and personalized treatment.

Leaky blood vessels are a hallmark of solid tumors. However, the molecular mechanisms and clinical implications of vascular leakage in human cancer remain unexplored. Here, we identified fibrinopeptide-A (FpA) as a robust in-situ marker of vascular leakage, analyzing diagnostic specimens from two non-small cell lung cancer (NSCLC) cohorts (N=327 and N=200). Mechanistically, FpA+ staining localized to discrete stromal niches characterized by increased endothelial VEGF receptor-2 phosphorylation, elevated VEGFA production by tumor cells and loss of the endothelial tyrosine phosphatase PTPRB. Immune profiling revealed reduced density of mature dendritic cells and granzyme B-expressing cytotoxic T cells in tumors with high-leakage. This leakage-associated immunosuppression was linked to the presence of tertiary lymphoid structures (TLS) in lung adenocarcinoma (LUAD). However, compared to low-leakage, TLS in high-leakage tumors were enriched in regulatory T cells and conferred no survival advantage. Patients with high vascular leakage exhibited a reduced overall survival. Moreover, in a separate immunotherapy cohort (N=64), high-leakage was associated with poor response to anti-PD-1/PD-L1 treatment. This study establishes vascular leakage as an important prognostic factor in NSCLC, and provides mechanistic understanding and methodological framework to stratify NSCLC patients with regard to responsiveness to immunotherapy.

Researchers have found that some lung tumors in mice can cause cachexia by directly communicating with neurons to alter eating behavior. Further, researchers showed that efforts to reduce cachexia through feeding the mice a high-fat diet paradoxically worsened the problem.

Lung cancer is strongly associated with increased extracellular matrix (ECM) stiffness, which correlates with poor patient prognosis. Our study reveals that high-stiffness tumor niches exhibit significant upregulation of neutrophil extracellular traps (NETs), which enhance the Warburg effect and promote tumor cell proliferation. Using atomic force microscopy (AFM) and multi-immunofluorescence staining (mIF), we demonstrated a spatial correlation between NETs formation and localized ECM stiffness in lung cancer tissues. In KrasLSL-G12D/+/Trp53 fl/fl transgenic mouse model, bleomycin-induced lung stiffening further increased NETs generation, while genetic ablation of Pad4 (Pad4-/- mouse model) or pharmacological inhibition of NETs (via LOX mAb, BAPN, or DNase I) reduced tumor burden. Mechanistically, stiffness-driven NETs upregulated glycolytic enzymes and extracellular acidification rate (ECAR) through aberrant mTOR pathway activation. Ex vivo and patient-derived xenograft (PDX) models validated the therapeutic potential of targeting LOX could extracellularly attenuate stiffness of ECM and intracellularly inhibit mTOR pathway. Our findings propose a novel strategy to improve lung cancer outcomes by disrupting the stiffness-NETs-mTOR axis, offering a one target-dual function approach for tumors with stiffening ECM.

Lung adenocarcinoma (LUAD) remains a leading cause of cancer mortality. Although targeted therapies and immunotherapies have improved outcomes, many patients exhibit limited responses due to primary or acquired resistance, underscoring the need to identify novel molecular targets. The Gab family of scaffolding adaptors, including GAB1 and GAB2, are recognized oncogenic regulators, whereas the role of GAB3, implicated in immune cell activation, is poorly defined in solid tumors. Here, we identify GAB3 as a novel tumor suppressor in LUAD. Pan-cancer analysis revealed frequent GAB3 downregulation, and high GAB3 expression was associated with favorable prognosis. Functionally, GAB3 overexpression suppressed LUAD cell proliferation, migration, invasion, and tumor growth in vitro and in vivo. Mechanistically, GAB3 interacted with LYN kinase to inhibit the MAPK signaling pathway and reverse epithelial-mesenchymal transition (EMT). In addition, GAB3 remodeled the tumor immune microenvironment, enhanced CXCL10 secretion, increased CD8+ T cell infiltration and effector function, and potently sensitized tumors to anti-PD-1 therapy. Our findings support a dual tumor-suppressive mechanism for GAB3 and propose it as a promising prognostic biomarker and therapeutic target in LUAD.

Non-small cell lung cancer (NSCLC) continues to pose a significant global health challenge, as conventional therapies often exhibit limited effectiveness. Ferroptosis has emerged as a critical vulnerability across diverse malignancies, including NSCLC, offering a promising opportunity for therapeutic intervention. Nevertheless, molecular mediators dictating ferroptosis sensitivity and exploitable vulnerabilities remain largely elusive in NSCLC. In this study, pyruvate carboxylase (PC) is identified as a pivotal ferroptosis regulator that is highly expressed in NSCLC tissues and associated with aggressive progression and poor prognosis. Functionally, PC promotes NSCLC cell proliferation, migration, and invasion both in vitro and in vivo. Mechanistically, PC confers resistance to ferroptosis by suppressing AMPK activation in an ATP-dependent manner, thereby activating the AKT-mTOR signaling pathway, upregulating fatty acid synthase (FASN) and malonyl-CoA-acyl carrier protein transacylase (MCAT), and ultimately inhibiting the PPARα-ACSL4 axis. Further analyses revealed that PC is targeted by Deltex E3 ubiquitin ligase 1 (DTX1) and ubiquitin-specific peptidase 51 (USP51), which modulate NSCLC progression by altering ferroptosis susceptibility. Therapeutically, the combination of the USP51 inhibitor dihydromyricetin (DHM) and anti-PD-1 treatment enhances antitumor efficacy with increased ferroptosis in vivo. Collectively, these results demonstrate that blocking PC facilitates ferroptosis, highlighting PC as a potential therapeutic target for NSCLC.

Tumor-infiltrating plasma cells are key responders to immunotherapy. In this issue of Cancer Cell, Meyerhoff et al. show that plasma cells from lung lesions of anti-PD-1-treated patients target citrullinated proteins. Engineering CAR T cells with these plasma cells scFv reveals anti-tumor specificity without adverse effects, identifying therapeutic opportunities.

The role of tumor-infiltrating B cells (TIL-Bs) in shaping anti-tumor responses in the context of immune checkpoint blockade remains incompletely understood. Here, we interrogate the humoral response in resected lung tumors from patients with non-small cell lung cancer (NSCLC) treated with neoadjuvant PD-1 blockade. We find that tumors orchestrate tertiary lymphoid structures with CD138+ plasma cells, from which we clone recombinant monoclonal antibodies (mAbs) using B cell receptors (BCRs) exhibiting somatic hypermutation and class switching. Several mAbs bind cell-surface citrullinated proteins, characteristic of cancer cells. Chimeric antigen receptor (CAR) T redirected with the soluble chain fragment variable (scFv) of our lead candidate antibody (PC-1) specifically target tumor cells and tumor-promoting myeloid cells in vivo without off-target activity. Moreover, ablation of the citrullination enzyme PADI2 in tumor-bearing mice eliminates reactivity to PC-1 and cytotoxic killing by the CAR. Our results implicate a therapeutic potential for tumor-infiltrating plasma cells that may be harnessed for cancer treatment.

Small molecules that modulate protein complexes have transformed cell biology and oncology, yet few chemical starting points exist to probe protein-protein interactions. To expand this space, we developed molecular COUPLrs, elaborated small molecules flanked by two cysteine‑reactive warheads. Using CONNECT, an integrated chemical proteomic platform that identifies proteins and complexes amenable to coupling, we revealed 171 targetable protein classes, including mutant‑selective complexes and assemblies not traditionally addressed by small molecules. We then optimized a COUPLr against the oncogenic fusion EML4‑ALK. This compound engages EML4‑ALK by binding its EML4 domain, remodeling protein dynamics, disrupting downstream signaling, and inducing proteasome‑mediated degradation of the fusion. Finally, we show that FDA‑approved drugs can be converted into COUPLrs to degrade their targets, indicating that this modality can endow existing therapeutics with new functional properties. Overall, molecular COUPLrs offer an unbiased framework to discover, characterize, and pharmacologically exploit protein complexes.

Lung cancer remains a leading cause of cancer-related mortality worldwide, with metastatic disease accounting for ∼90% of deaths. While p53 gene therapy offers promise for restoring tumor suppressor function, its clinical translation has been hindered by inefficient delivery systems and pulmonary administration challenges. Here, we developed redox-responsive thiolated chitosan nanoparticles (TCS NPs) as the first inhalable nanotherapeutic platform for tumor-selective p53 gene delivery to treat metastatic lung cancer. Our system exploits elevated glutathione levels in cancer cells to trigger selective gene release, while mucoadhesive properties ensure prolonged pulmonary retention. The p53 DNA-loaded TCS NPs demonstrated exceptional cancer selectivity, exhibiting significant cytotoxicity against lung cancer cells while maintaining biocompatibility with normal tissues. Treatment restored p53-mediated pathways, upregulating metastasis suppressors (KLF6 and E-cadherin) and apoptotic factors (Bax and cleaved caspase-3), thereby inhibiting migration, invasion, and metastatic progression. In vivo studies using ectopic xenograft and experimental metastasis models confirmed robust therapeutic efficacy, with inhaled TCS@p53 NPs achieving significant tumor suppression and reduced metastatic nodule formation while maintaining excellent safety profiles. This work presents a transformative inhalable gene therapy platform that overcomes traditional delivery limitations through tumor-selective targeting.

Interleukin-1β (IL-1β) has long been characterized as a foundational mediator of innate immunity, yet this designation captures only a fraction of its biological significance. A growing body of evidence now positions IL-1β as a molecular arbiter of tissue restoration - a context-sensitive instructor that orchestrates the dynamic interplay between inflammatory signaling, stem cell fate, and niche remodeling that collectively determines whether an injured tissue regenerates or degenerates. Far beyond its canonical role in host defense, IL-1β functions as a critical regulator of cell state plasticity, driving the transition of lineage-committed progenitors into highly plastic, transitional identities that are indispensable for effective wound repair. Yet the very mechanisms that render IL-1β essential for regeneration also render it capable of profound pathological consequence when its activity is sustained, amplified, or spatiotemporally dysregulated. This review explores the spatiotemporal logic governing IL-1β-mediated plasticity, with particular emphasis on the respiratory epithelium as a primary and instructive model of inflammatory reprogramming and niche remodeling. We examine how the signaling thresholds, cellular contexts, and temporal dynamics of IL-1β activity collectively determine regenerative outcomes - and how the failure to terminate this program pathologically stabilizes transitional progenitor states, predisposing tissues to fibrosis, functional exhaustion, and malignant transformation. By integrating evidence spanning transcriptional, epigenetic, and metabolic rewiring, we propose a unified conceptual framework for understanding IL-1β as a master coordinator of the regenerative ecosystem - one whose precise modulation represents a compelling therapeutic target for resetting pathological cell states toward functional recovery in chronic inflammatory diseases.

Distant metastasis, characterized by organotropism, is a major cause of mortality in lung adenocarcinoma (LUAD). In this study, digital spatial profiling (DSP), multiplex immunofluorescence (mIF), and clinical data from 52 LUAD patients were integrated to develop organ-specific metastasis risk models, and the molecular mechanisms underlying metastatic organotropism were investigated. Random forest models based on primary tumor spatial transcriptomics accurately predicted metastasis to the brain (AUC = 0.974), liver (AUC = 0.975), adrenal gland (AUC = 0.929), and bone (AUC = 0.907). Key compartment-specific gene expression signatures associated with organotropic metastasis were identified, including those expressed in tumors (e.g., FKBP1A for the brain and MOCOS for the liver), immune cells (e.g., ADAMTSL2 for the liver), and stromal cells (e.g., CKAP2 for the brain). Pathway analyses revealed distinct biological processes associated with organotropism, such as enriched cell death pathways in brain metastasis and extracellular matrix (ECM) remodeling in liver metastasis. Postmetastasis survival models highlight stromal gene expression (e.g., PKM for OS and VCAM1 for PFS) and immunosuppressive microenvironments (e.g., M2 macrophage infiltration) as critical prognostic factors. The high-precision prediction models and key molecular signatures identified in this study enhance our understanding of "seed-soil" interaction dynamics and offer promising biomarkers and therapeutic targets for future clinical use.

MicroRNA (miRNA) silencing is classically ascribed to RNA-sequence rules that guide Argonaute 2 (AGO2) targeting. Using chimeric eCLIP and complementary analyses in CRISPR-edited human lung epithelial cells, we show that efficient miRNA targeting also depends on the AGO2 adaptor-scaffold LIMD1. In LIMD1-deficient cells, AGO2 binds more miRNAs, but each AGO2-miRNA engages fewer transcripts and sites, reducing occupancy and halving both the breadth and depth of targeting. LIMD1 dependence is most pronounced for poorly conserved, weakly seed-paired sites that nonetheless form stable duplexes. LIMD1 deficiency alters AGO2 footprints and derepresses oncogenic targets inversely correlated with LIMD1 expression in lung adenocarcinoma. Thus, LIMD1 modifies the outcome of sequence-defined interactions that would otherwise be infrequent, unstable, or unproductive, revealing an adaptor-governed layer of posttranscriptional regulation beyond RNA-sequence rules.

Lung cancer remains the leading cause of cancer-related death worldwide, and long noncoding RNAs (lncRNAs) have been implicated in its tumourigenesis and progression. However, the roles lncRNAs play in lung cancer remain unclear. In this study, we discovered an important role for the lncRNA MNX1 Antisense RNA 1 (MNX1-AS1) as a critical regulator of one-carbon metabolism reprogramming. Glutamine depletion altered chromatin accessibility, leading to downregulation of MNX1-AS1, while elevated expression of MNX1-AS1 was correlated with poor prognosis in patients with non-small cell lung cancer. Functional studies showed that MNX1-AS1 promoted cell proliferation and sphere formation in vitro, and subcutaneous and orthotopic tumour growth in vivo. Mechanistically, MNX1-AS1 directly binds to calcyclin binding protein (CACYBP), protecting it from ubiquitin-mediated degradation; thus, the MNX1-AS1/CACYBP complex accelerates the transcription of key one-carbon metabolism-related genes through the Wnt/β-catenin pathway. MNX1-AS1/CACYBP/β-catenin axis upregulated key one-carbon metabolism-related genes, which were essential for generating related metabolites and maintaining cellular redox balance to support lung cancer cell proliferation. These findings established that the lncRNA MNX1-AS1 acts as a crucial driver of one-carbon metabolism reprogramming in non-small cell lung cancer and highlight that the newly identified MNX1-AS1/CACYBP/β-catenin axis may serve as a potential prognostic biomarker and therapeutic target for lung cancer intervention.

Limited intratumoral persistence and insufficient proliferative capacity severely restrict the efficacy of chimeric antigen receptor (CAR)-T cell therapies in solid tumors. Here, we demonstrated that DLL3-targeting CAR-T cells co-expressing a CD56 chimeric switch receptor (CSR) and incorporating parallel 4-1BB costimulatory signaling (DBBζ.CBB) effectively address these limitations. In preclinical small cell lung cancer (SCLC) models, DBBζ.CBB exhibited sustained tumor infiltration, prolonged persistence, and superior antitumor activity. Mechanistically, parallel 4-1BB signaling dynamically programed CAR-T cell fate by promoting early expansion and memory maintenance, driving a highly proliferative effector state at the intermediate stage, and delaying terminal exhaustion at the later stage, thereby sustaining in vivo persistence and enabling durable antitumor responses. Building upon the intratumoral T-cell pool established by DBBζ.CBB, subsequent DLL3 trispecific T-cell engager (TriTCE) administration synergistically enhanced tumor eradication by further boosting CD8+ T cell infiltration and overall activation while mitigating exhaustion and terminal differentiation. Collectively, these findings establish a clinically translatable combinatorial framework to enhance the efficacy and durability of CAR-T therapy in solid tumors.

Angiogenesis fuels cancer progression, yet its immunomodulatory role in neuroendocrine malignancies remains poorly understood. Recently, Wang et al. identified a blood-brain barrier-like vascular gate that mediates immune exclusion and immunotherapy resistance in small-cell lung cancer. Targeting this barrier with OSI-906 enhances CD8+ T-cell infiltration and immunotherapy efficacy.

Although significant progress has been made in identifying antigen targets in autoimmune encephalitis (AIE), in a substantial proportion of patients with presumed AIE no known autoantibodies can be detected. Herein, we describe a novel autoantibody reactivity, directed against copine-5 (CPNE5), in patients with AIE and melanoma. Patients were identified through routine clinical testing for anti-neural autoantibodies by indirect immunofluorescence on neuronal tissue sections. The antigen was identified by immunoprecipitation and mass spectrometry and confirmed by recombinant protein assays. Serum and cerebrospinal fluid (CSF) immunoglobulin G (IgG) from four patients was found to exhibit distinct binding to cerebellar and hippocampal neurons. Copine-5 was identified as the target antigen. Clinical manifestations included cognitive decline, confusion/disorientation, psychosis, seizures and signs and symptoms compatible with basal ganglia involvement (movement disorders, including secondary parkinsonism, and/or abulia/akinetic mutism), alongside pain (including painful tonic spasms and focal allodynia), dysarthria, dysphagia, and abducens nerve palsy. Brain magnetic resonance imaging disclosed hyperintense lesions in the basal ganglia and the temporal lobe. CSF analysis revealed mild pleocytosis, intrathecal IgG synthesis, and blood/CSF barrier dysfunction. Of particular note, all four patients had melanoma, with an occult primary in three. In one patient, melanoma diagnosis and immunotherapy with the anti-PD-1 immune checkpoint inhibitor nivolumab preceded the onset of AIE by 9 months. Glucocorticoid treatment and/or intravenous immunoglobulins led to transient improvement in at least three patients; however, all relapsed and three progressed to hypoactive delirium or mutism. Plasma exchange and cyclophosphamide treatment were followed by clinical improvement and stabilization in one patient, but potentially contributed to fatal infectious complications. Copine-5-IgG belonged to the strongly complement-activating IgG1 subclass and was produced intrathecally. Serum titers ranged between 1:1000 and 1:100,000. Copine-5 is a novel autoantibody target in AIE. Testing for anti-copine-5 autoantibodies should be included in the diagnostic workup of patients with AIE, especially, but not exclusively, if associated with melanoma or positive melanoma-associated tumor markers. Further studies investigating the immunopathogenesis of copine-5-related autoimmunity and the potential significance of anti-copine-5 as a novel paraneoplastic serological marker and of copine-5 as a histopathological tumor marker in patients with suspected melanoma are highly warranted.

Nearly 11 years after T-VEC became the first oncolytic virus approved in a major market, the closest candidate to follow it-Replimune's RP1, now facing an FDA advisory committee-remains a locally injected herpesvirus for melanoma, underscoring how little the field has advanced. Researchers blame a stack of unresolved obstacles-flawed trial designs, mistimed drug combinations, tumor defenses, such as hypoxia, and the inability to deliver most viruses intravenously-even as smarter sequencing, neoadjuvant approaches, and a bladder-cancer virus near approval hint at renewed momentum.

Melanomas display distinct transcriptomic states, but it remains unclear how they associate with clinical outcomes. We performed digital spatial RNA profiling (DSP-RNA) of metastatic tumors from patients to investigate how transcriptomic states correlate with melanoma specific survival (MSS) and acral melanoma (AM). We performed DSP-RNA across a tissue microarray constructed from 111 patients with in-transit metastatic melanoma (ITM) diagnosed from 1990 to 2020. Data quality control, noise correction, and normalization yielded high-quality profiles from 105 patients, including 30 (36%) who received immune checkpoint inhibitors and 20 (24%) with AM. We performed principal component (PC) analysis and correlated the results with published gene signatures: The PC1 axis differentiated transitory from undifferentiated melanoma, PC2 reflected immune cell infiltration, PC3 corresponded to stromal cells and neural crest-like melanoma, and PC4 associated with melanocytic melanoma. Across a cohort of treatment-naïve ITM, high expression of the melanocytic state conferred a median MSS difference of 7.72 years (melanocytic "high" = 5.16 years versus "low" = 12.88 years, log-rank P = 0.0061) and independently associated with poor survival in multivariate analysis. AMs showed higher melanocytic state gene expression compared to nonacral. Findings were validated in external datasets, supporting that the melanocytic state predicts poor prognosis. The melanocytic state is associated with poor prognosis and may be enriched in AM, implying that identifying patients with melanocytic melanoma may be important for therapeutic decisions. Unlike other gene expression predictors proposed for prognostic stratification, the melanocytic state characterizes a biological subtype, suggesting that it may have specific therapeutic vulnerabilities.

Flat head/neck pigmented macules can be diagnostically challenging. To assess the impact of reflectance confocal microscopy (RCM) integration to dermoscopic triage of equivocal lesions. Prospective multicentre study of equivocal flat macules. Lesions suspicious or discordant were biopsied/excised; double-negative lesions were followed ≥1 year. Histopathology served as reference standard. A total of 2006 lesions from 801 patients (mean age 66.2 years) were evaluated. After dermoscopy-RCM integration, 236/2006 (11.8%) lesions were managed as malignant (226 biopsied/excised; 10 noninvasively treated [8 BCC, 2 LM]). Twenty-six lesions changed at follow-up and were removed, with 4 malignancies (0.4%; 3 LM, 1 BCC). Histology (n=252) confirmed 114 malignancies (62 LM/LMM; 52 BCC/SCC). Dermoscopy detected 54.8% of LM/LMM and 90.4% of BCC/SCC; RCM correctly diagnosed 91.9% of LM/LMM and 94.2% of BCC/SCC. Verification and selection biases and follow-up losses. Integrating RCM significantly improves LM/LMM sensitivity detection, and specificity for benign lesions compared to dermoscopy alone, minimizing overtreatment and missed malignancies. Although a small residual risk of missed malignancy persists, the combined approach offers a highly safe, non-invasive pathway for managing equivocal lesions in cosmetically sensitive areas.

Intentional outdoor tanning involves exposing one's skin to UV radiation from the sun for the purpose of darkening the skin to create a tanned appearance. Although robust surveillance data are available on many skin cancer risk factors, limited data on intentional outdoor tanning prevalence have been reported to date. To examine the prevalence of intentional outdoor tanning among US adults and the factors associated with this behavior. This cross-sectional study used data from US adults who participated in the 2024 National Health Interview Survey, a nationally representative sample of civilian, noninstitutionalized US adults aged 18 years or older, to assess the prevalence of intentional outdoor tanning and sunburn by asking participants "when spending time outdoors, how often do you try to get some sun for the purpose of developing a tan?" (always, most of the time, sometimes, rarely, or never) and "during the past 12 months, did you have a sunburn even on a small part of your skin?" (yes or no). The individual-level factors analyzed were demographic characteristics and health-related behaviors, including physical activity and alcohol use. The primary outcome was prevalence of intentional outdoor tanning. The secondary outcome was prevalence of sunburn. A total of 31 615 US adults (mean [SD] age, 48.3 [18.6] years; 17 111 females [51.4%]) were included in the study. Overall, the unadjusted prevalence of intentional outdoor tanning among adults was 30.2% (35.6% [95% CI, 34.6%-36.7%] in women and 26.6% [95% CI, 25.6%-27.7%] in men). Among women and men, prevalence of intentional outdoor tanning was associated with age, race and ethnicity, sun sensitivity, physical activity, and alcohol use. Among women, prevalence was highest among those aged 18 to 29 years (48.1%; 95% CI, 45.3%-50.9%), non-Hispanic White women (41.3%; 95% CI, 39.9%-42.7%), non-sun-sensitive women (38.3%; 95% CI, 36.6%-40.1%), women who met both aerobic and strength guidelines (39.5%; 95% CI, 37.6%-41.5%), and women who engaged in heavy or binge drinking (47.2%; 95% CI, 44.8%-49.7%) (P for all < .001). Among men, prevalence was highest among those aged 18 to 29 years (29.9%; 95% CI, 27.4%-32.6%), non-Hispanic White men (30.5%; 95% CI, 29.0%-32.1%), sun-sensitive men (28.7%; 95% CI, 27.2%-30.2%), men who met both aerobic and strength guidelines (32.2%; 95% CI, 30.2%-34.3%), and men who engaged in heavy or binge drinking (31.2%; 95% CI, 29.0%-33.5%) (P for all < .001). Among women only, prevalence was associated with educational level and body mass index, with a higher prevalence among women with some college (37.5%; 95% CI, 35.8%-39.3%) and those at a healthy weight (38.7%; 95% CI, 37.0%-40.5%; P for all < .001). Among men only, prevalence was associated with marital status, with a higher prevalence among married or partnered men (25.8%; 95% CI, 24.5%-27.1%; P = .04). The prevalence of sunburn was significantly higher among adults who reported intentional outdoor tanning (49.9%; 95% CI, 48.5%-51.4%) compared with those who did not (28.3%; 95% CI, 27.4%-29.3%; P < .001). This cross-sectional study found that the highest prevalence of intentional outdoor tanning occurs among US women who are younger, non-Hispanic White, and physically active and report heavy and/or binge drinking. Those who engage in intentional outdoor tanning are more likely to experience sunburn, increasing their skin cancer risk. These findings can inform future strategies to address intentional outdoor tanning as part of larger efforts to reduce skin cancer risk.

Beyond their conventional role as passive transdermal delivery vehicles, microneedle (MN) platforms now function as active bio-interfaces capable of modulating therapeutic responses in both localized and systemic diseases. This review summarizes recent advances in MN technology, focusing on the transition from traditional matrix-controlled delivery to bioactive microneedles. Although localized applications at barrier surfaces, such as treating cutaneous disorders and mucosal lesions, remain a fundamental focus, this review emphasizes the application of MNs in complex chronic metabolic diseases (e.g., diabetes), oncology (e.g., melanoma and glioblastoma), and deep-tissue degenerative diseases of the cardiovascular, nervous, and musculoskeletal systems. Integrating stimuli-responsive materials, including metal-organic frameworks (MOFs), aggregation-induced emission luminogens (AIEgens), and smart hydrogels, with external physical stimuli enables autonomous, closed-loop interventions, thereby advancing personalized systemic therapy. Furthermore, we summarize recent progress in applying MNs to non-traditional sites and deep-tissue repair. Finally, rather than focusing solely on phenotypic efficacy, we discuss key translational challenges, including manufacturing scalability, biosafety, and regulatory pathways, to guide future clinical translation.

The efficacy of dendritic cell (DC) cancer vaccines is linked to poor immunogenicity of tumor-associated antigens and failure to elicit robust MHC class II-restricted CD4⁺ T-cell responses. Here, we introduce PROTEXI, a DC vaccine platform that optimizes tumor immunity by co-presenting tumor-specific CD8⁺ T-cell epitopes alongside CD4⁺ T helper epitopes from the SARS-CoV-2 Spike protein, leveraging widespread anti-viral immunity. In preclinical mouse models of melanoma and breast cancer, PROTEXI significantly reduces tumor growth and improves survival by promoting robust T cell infiltration into immune-cold tumors, increasing cytotoxic T cell responses via epitope spreading, and activating genes linked to optimal DC, NK cell, and T cell function. Furthermore, PROTEXI elicits superior responses when combined with other immunotherapy agents in models of therapy-resistant tumors. Finally, in a humanized mouse model of melanoma, PROTEXI vaccine co-presenting CD4⁺ T-specific Spike epitopes with CD8⁺ T cell-restricted PRAME and MAGE-A3 antigens, significantly reduces tumor burden. Thus, these data underscore the potential of harnessing pre-existing viral-specific immunity to enhance the efficacy of DC vaccines in immune-cold tumors.

Translating preclinical research findings to clinical applications presents challenges due to the metabolic and anatomical disparities between animal models and humans. Although organ-on-a-chip models replicate the organization of human cells, they lack the complexity of the microenvironment and anatomical fidelity. Developing alternative research models with greater physiological and anatomical relevance to humans holds the potential to enhance translational success and reduce reliance on animal experimentation. In this study, we have showcased the technical details and conditions to maintain the viability of ex vivo perfused large human abdominal fasciocutaneous flaps for up to three weeks. Our efforts involved refining surgical procedures, vascular territory mapping (angiosome analysis), and pedicle exploration; engineering the bioreactor system; and optimizing perfusion media. Angiography via thermal and fluorescent methods was used to confirm the perfusion success. Metabolic activity was closely monitored by tracking glucose consumption and lactate production. Assessments of tissue viability encompassed histological analysis, TUNEL staining, gene expression profiling, measurement of vascular and metabolic reactivity, and in vitro propagation of isolated adipose stem cells and dermal fibroblasts. Furthermore, we harnessed our optimized human skin perfusion model to investigate the dynamics of radiation and chemical-induced injuries. Additionally, we explored the model's utility in studying adipose tissue metabolism and employed human skin tissue to establish melanoma and breast cancer tumor models. To our knowledge, this is the first model capable of preserving the function and viability of a large flap for extended periods, providing a proof-of-concept foundation for diverse research applications.

Since 2012, brown bullhead catfish (Ameiurus nebulosus) in a lake that spans Vermont, USA, and Quebec, Canada, have shown a high rate of melanomas, suggesting a causal contaminant or contagion1. We tested the hypothesis that this affliction represents a clonally transmissible cancer, a rare phenomenon in which cancer cells themselves spread between individuals, behaving more like parasites than conventional tumours2. Whole-genome sequencing of tumour and matched non-tumour host tissues revealed that tumour mitochondrial and nuclear genomes are more closely related to each other than to their hosts or unaffected fish. Hundreds of thousands of genetic variants are shared among tumour samples but are absent from host fish, vastly exceeding levels seen in conventional cancers3. These findings indicate that melanoma in these brown bullheads represents the fourth documented type of naturally occurring transmissible cancer in animals, after dogs4, Tasmanian devils5 and several bivalve species6-13. This raises important questions about the cancer's origin, the mode of transmission and the long-term impact on fish populations.

Therapy-induced senescence (TIS) is a common outcome of diverse anticancer treatments, including chemotherapy, radiation, and small-molecule inhibitors. Senescence is characterized by stable growth arrest and the senescence-associated secretory phenotype (SASP), which includes various immune mediators. As the role of the immune system in controlling cancer becomes increasingly appreciated, understanding the impact of TIS on the tumor immune microenvironment (TIME) is critically important. Here, we investigated how senescence can be leveraged to enhance antitumor immune responses. We investigated the effects of an Aurora kinase A inhibitor (AURKAi), a potent inducer of senescence in melanoma models, using transcriptome and secretome profiling. The role of the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway was investigated using imaging, inhibitors, and gene knockout. We also examined the effect of AURKAi on the surface expression of major histocompatibility complex class I (MHC-I) and programmed death-ligand 1 (PD-L1), as well as on signal transducer and activator of transcription 1 (STAT1) activation. In vivo, the effects of AURKAi treatment on the TIME were investigated using spectral cytometry and cell depletion studies. Finally, we assessed combining AURKAi with immune checkpoint blockade (ICB), adoptive cell therapy, and natural killer (NK) cell therapy in murine models. We observed significant gene expression rewiring and the secretion of immune-related mediators and chemokines associated with AURKAi-induced senescence, which was favorably modulated by a senolytic BCL-2/xL inhibitor navitoclax. Mechanistically, AURKAi induced the formation of micronuclei linked with the activation of the cGAS-STING pathway, which, in turn, initiated pro-inflammatory transcriptional programs. Senescent melanoma cells exhibited increased surface expression of MHC-I and PD-L1, along with interferon regulatory factor 3 (IRF3) and STAT1 activation, indicating enhanced immunogenicity. In vivo, AURKAi treatment significantly enriched the TIME with activated CD8+ T cells and NK cells, with depletion studies confirming their critical role in antitumor effects. While combining AURKAi with immune checkpoint blockade was not beneficial, AURKAi pretreatment significantly augmented T and NK cell therapies in murine models, resulting in enhanced tumor control and prolonged survival. These findings suggest that senescence, specifically when induced by an AURKAi, can be harnessed to promote effector cell engagement in immune-cold tumors, thereby enhancing responsiveness to cell therapies.

To evaluate the clinical applicability of previously established transcriptomic signatures (molecular subtypes, components and GemPred status) in metastatic pancreatic cancer, we conducted a retrospective pooled analysis of 178 patients from three phase 2 trials (PRODIGE35/37, AFUGEM; 2013-2016) testing first-line regimens (FOLFIRINOX, GemNab, FuNab, FOLFIRI3). RNA sequencing was performed on primary/metastatic tumors across French centers, with blinded assessment of subtypes (immune classical, pure basal-like, stroma-activated), quantitative components, and GemPred status. Primary endpoint: progression-free survival (PFS). Immune classical subtype showed superior median PFS (9.03 months) and OS (11.27 months) versus basal-like and stroma-activated subtypes (PFS: p=0.015; OS: p=0.010). Higher classical component correlated with improved PFS (HR=0.83, p=0.048) and OS (HR=0.71, p=0.001). Inactive stroma predicted better PFS (HR=0.64, p=0.001) and OS (HR=0.71, p=0.015). GemPred-negative patients treated with FOLFIRINOX versus GemNab had higher ORR (46.9% vs. 19.1%, p=0.046), longer PFS (8.2 vs. 2.3 months; HR=2.28, p=0.008), and OS (11.6 vs. 5.0 months; HR=2.04, p=0.021). No differences occurred in GemPred-positive patients. In a formal treatment-by-GemPred interaction analysis (FOLFIRINOX vs. GemNab), the interaction was significant for OS (adjusted p-interaction=0.050) but not for PFS (adjusted p-interaction=0.51). Transcriptomic signatures retain prognostic and predictive utility in metastatic pancreatic cancer, with GemPred representing a hypothesis-generating predictive signal for OS (e.g., a FOLFIRINOX OS benefit in GemPred-negative). Prospective validation is warranted for clinical implementation.

New-onset diabetes (NOD) increases the risk of pancreatic cancer. Previous studies have mainly focused on the presence or absence of diabetes, with limited attention to NOD severity at diagnosis and its clinical course. This 15-year longitudinal nationwide cohort study aimed to analyse the pancreatic cancer risk based on the initial severity and progression pattern of NOD. We included 402 663 individuals with or without NOD from the Korean National Health Insurance Service (2005-2019). Initial NOD severity was defined by baseline antidiabetic treatment intensity: no antidiabetics, oral antidiabetics and insulin. Pancreatic cancer risk increased stepwise with higher initial treatment intensity in NOD compared with individuals without diabetes (adjusted HRs (aHRs) 3.01, 4.32 and 5.60 for no antidiabetics, oral antidiabetics and insulin, respectively). Within the same baseline treatment-intensity category, individuals with rapid escalation within 6 months had a higher risk of pancreatic cancer than those with stable or decreased treatment intensity. In some comparisons between groups with different baseline treatment intensities, the risk of pancreatic cancer was higher with less intensive baseline therapy but subsequent intensification than with more intensive baseline therapy without progression. Notably, drug-naïve individuals who initiated antidiabetic treatment within 6 months had a greater risk even than those who were initially on oral medication without worsening (aHR 6.50 vs 3.67). These patterns were more prominent in pancreatic cancer cases diagnosed within 3 years after NOD. Greater initial severity of NOD and rapid early aggravation were both associated with an increased risk of pancreatic cancer.

Pancreatic cancer continues to be among the most lethal malignancies. The distinct tumor microenvironment (TME) not only facilitates tumor cell proliferation, invasion, and metastasis but also establishes a formidable barrier to the diffusion of chemotherapeutic agents and immunotherapies. Consequently, it is imperative to identify novel compounds that target the tumor microenvironment (TME). This study aims to identify a small molecule as a therapeutic agent for pancreatic cancer through dual mechanisms, specifically DDR1 inhibition and methuosis induction. We characterized DDR1 as a primary target of N-(4-Methyl-3-(4-(5-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-pyrazol-1-yl)phenyl)-6-(trifluoromethyl)picolinamide (IHMT-140) using ADP-Glo assays, KINOMEscan, Western blotting, and molecular docking. Colony formation, invasion, wound-healing migration, and immunohistochemical staining assays demonstrated that IHMT-140 suppresses epithelial-mesenchymal transition (EMT) by regulating key EMT markers. Cell viability assays, including Cell Titer-Glo and live-cell imaging, transmission electron microscopy (TEM), and immunofluorescence staining, revealed that IHMT-140 triggers methuosis in pancreatic cancer cells. RNA-seq was also employed to confirm the underlying mechanisms. Finally, Cell Titer-Glo, Western blot, and flow cytometry were applied to demonstrate the combined antitumor effects of IHMT-140 and gemcitabine. Gemcitabine uptake and tumor accumulation were quantified by high-resolution mass spectrometry. The in vivo efficacy was tested in a xenograft model, with tumor sections analyzed by H&E and pan-collagen staining. We discovered N-(4-Methyl-3-(4-(5-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-pyrazol-1-yl)phenyl)-6-(trifluoromethyl)picolinamide (IHMT-140) as a novel dual-function agent that acts as a DDR1 inhibitor and a methuosis inducer. It simultaneously remodels the tumor stromal ECM and activates excessive macropinocytosis. Pharmacological evaluation shows that IHMT-140 inhibits tumor invasion and metastasis, enhances gemcitabine accumulation in tumors, and significantly improves therapeutic outcomes. Our findings present a promising therapeutic strategy for pancreatic cancer, integrating DDR1 kinase inhibition with methuosis induction to augment chemotherapy efficacy. This approach provides a potential novel approach for pancreatic cancer therapy.

Pancreatic adenocarcinoma (PAAD) is a highly lethal malignancy with limited prognostic biomarkers and therapeutic targets. Lactate-driven lactylation has recently emerged as an important regulator of cancer progression, but its role in PAAD remains unclear. In this study, integrative analysis of TCGA and GEO datasets, combined with experimental validation, identified a five-gene lactylation-associated signature (LRP3, TTLL6, TSGA13, PRKCG, and SDK2) that effectively stratified PAAD patients by survival risk. High-risk tumors displayed an immunosuppressive phenotype with reduced immune infiltration, Th2-skewed remodeling, checkpoint activation, and distinct mutational and drug-sensitivity features. Among the signature genes, PRKCG was significantly downregulated in PAAD and associated with advanced disease and worse prognosis. PRKCG overexpression inhibited tumor cell proliferation, migration, invasion, and xenograft growth, while enhancing apoptosis. Mechanistically, lactate-induced lactylation impaired PRKCG-dependent activation of the p53 pathway without altering PRKCG expression, and mutation of predicted lactylation sites partially rescued this effect. These findings define a lactylation-associated prognostic model for PAAD and highlight the lactate-PRKCG-p53 axis as a potential therapeutic vulnerability.

Intravital microscopy (IVM) using dorsal skinfold chambers (DSCs) enables real-time, high-resolution imaging of the tumor microenvironment. Conventional, metal-based DSC systems often cause animal distress and require technically demanding surgical implantation. To address these challenges and adhere to the 3Rs principles of animal welfare, this study presents a novel, stereolithography (SLA)-based, 3D-printed DSC featuring a lightweight (0.69 g) biocompatible resin structure and a suture-free "one-click" fixation system. This affordable device simplifies surgical installation and minimizes postoperative inflammation, allowing continuous multimodal imaging-IVM, micro-CT, ultrasound, and in vivo fluorescence-for up to four weeks. The system is validated in mice bearing subcutaneous tumors generated from PDAC93-GFP pancreatic tumor organoids implanted in the DSC. Longitudinal, high-resolution imaging successfully tracks tumor expansion, active stromal remodeling characterized by progressive collagen fiber compaction and alignment, and the emergence of a highly tortuous peritumoral vascular network. Furthermore, in vivo tracking in Catchup mice reveals a distinct transition of tumor-associated neutrophils from random acute inflammatory migration to directed tumor-driven chemotaxis. Overall, this ergonomic and robust DSC design provides a highly reliable platform for sustained, high-quality assessment of tumor-stroma-immune interactions and therapeutic responses, while notably improving animal welfare.

Pancreatic ductal adenocarcinoma (PDA) is among the deadliest malignancies, driven by metastatic progression and profound cellular heterogeneity. We previously identified glutathione S-transferase theta 1 (GSTT1) as a regulator of a slow-cycling, highly metastatic tumor cell population, suggesting that GSTT1High cells may possess stem-like properties. Here, we define the functional and molecular features of this subpopulation in metastatic PDA. Using a mCherry-tagged Gstt1 reporter system in metastatic murine PDAC cells, we enriched for Gstt1High cells and observed increased tumor sphere formation, accompanied by upregulation of stemness-associated genes including PROM1 (CD133) and activation of Wnt and FGF signaling pathways. In human PDA models, CD133HighGSTT1High cells exhibited enhanced tumor sphere initiation and expansion compared to other populations, defining a maximal stem-like state. Notably, sensitivity to FGFR inhibitors was observed only under tumor sphere conditions, highlighting a context-dependent therapeutic vulnerability. Mechanistically, FGFR3 expression correlated with GSTT1 and CD133 levels, and FGF signaling was required to sustain this state. GSTT1 knockdown reduced CD133 protein levels, impaired tumor sphere formation, and altered sensitivity to FGFR inhibition. These findings were largely recapitulated in patient-derived PDA organoids, where GSTT1 and PROM1 co-expression predicted increased tumor sphere formation and enhanced response to the multi-kinase inhibitor Nintedanib. Together, these results identify a GSTT1HighCD133High stem-like subpopulation in metastatic PDA and identify an FGFR-dependent signaling axis that sustains this state, representing a potential therapeutic vulnerability.

Ferroptosis links cellular metabolism to immune regulation. Beyond its role as an iron-dependent form of regulated cell death, ferroptosis generates signals, including oxidized lipids, iron metabolites, and damage-associated molecular patterns, that influence inflammatory and immune responses. The pancreas is particularly susceptible to ferroptotic stress because of its high metabolic demand and close integration with immune and stromal networks. In pancreatitis, ferroptosis translates metabolic injury into innate immune activation, contributing to sterile inflammation and tissue damage. In pancreatic cancer, ferroptotic vulnerabilities can be exploited therapeutically, yet ferroptosis-associated signals may also support immune suppression, immune evasion, and treatment resistance. These findings suggest that ferroptosis functions as an immunometabolic checkpoint rather than simply a cell death program. Here, we discuss how ferroptosis shapes immune responses in pancreatitis and pancreatic cancer and examine the factors that determine whether it promotes inflammation, antitumor immunity, or immune tolerance. We also review ferroptosis-targeted therapies and the challenges associated with their clinical application.

Pancreatic ductal adenocarcinoma (PDAC) presents a substantial challenge due to its resistance to cancer treatments. This limited efficacy is, in part, attributed to the immunosuppressive tumor microenvironment (TME), which impairs effector T (Teff) cell activity. Interleukin-2 (IL-2) is a key cytokine for T cell activation, but its therapeutic use is limited by a short half-life, systemic toxicity, and regulatory T (Treg) activation. To address this limitation, we engineered Bifidobacterium longum, a probiotic obligate anaerobe that selectively colonizes the TME, to continuously secrete Super-mutant IL-2 (SumIL-2), an engineered IL-2 variant that preferentially activates Teff cells over Treg cells, thereby delivering SumIL-2 selectively to the tumor (BifidoSumIL-2). Systemic administration of BifidoSumIL-2 significantly suppressed tumor growth in both subcutaneous tumors and orthotopic PDAC in mice, inducing an improved Teff/Treg ratio. Combining BifidoSumIL-2 with chemotherapy, radiation, and immunotherapy further restrained orthotopic PDAC growth, highlighting its therapeutic potential for difficult-to-treat cancers like PDAC.

KRASG12D mutation drives oncogenic progression and creates an immunosuppressive microenvironment in cancers like pancreatic ductal adenocarcinoma and colorectal cancer. We investigate the immunomodulatory mechanisms of the KRASG12D inhibition and its synergy with natural killer (NK) cell therapies. We demonstrate that KRASG12D inhibition with MRTX1133 remodels the immune landscape by reducing myeloid-derived suppressor cell (MDSC) accumulation and facilitating infiltration and activation of NK and CD8+ T cells. Crucially, MRTX1133 reverses systemic immunosuppression, restoring the fitness of adoptively transferred NK cells. Mechanistically, KRASG12D inhibition impairs IFNGR1 palmitoylation and subsequent lysosomal degradation. MRTX1133 stabilizes IFNGR1 by reducing palmitoyltransferase expression and the palmitate pool. This stabilization increases IFN-γ/IFNGR signaling and up-regulates NK cell-activating ligands ICAM1 and ULBP1, thereby sensitizing cancer cells to NK cells. Consequently, combining MRTX1133 with IL-15 or adoptive NK cell therapy yields synergistic antitumor responses and prolonged survival. Our findings provide mechanistic rationale for combining KRASG12D inhibitors with NK cell-based immunotherapies to improve outcomes for patients with KRASG12D-mutant cancers.

The incidence of early-onset prostate cancer (EOPC) is rising, and by 2045 a 24.5% increase in cases and a 50% rise in mortality are projected. Accumulating evidence indicates that EOPC represents a distinct disease entity, characterized by unique molecular features, risk factor profiles, and clinical behavior that differ from standard-onset prostate cancer (SOPC). Nevertheless, research in this field remains nascent, and no consensus exists regarding the optimal management of EOPC. We synthesize current evidence on the epidemiology, molecular pathology, clinicopathological characteristics, survival, management, and early detection of EOPC. EOPC exhibits a distinctive molecular landscape, with TMPRSS2-ERG fusions occurring in 63-90% of cases as a hallmark alteration, whereas mutations in PTEN, SPOP, and CHD1 are significantly less frequent. Notably, the prevailing focus on hereditary EOPC has inadvertently led to the neglect of sporadic cases, which dominate clinical practice. Although localized EOPC confers no significant prognostic advantage over SOPC, high-risk or metastatic early-onset disease substantially elevates prostate-cancer-specific mortality. By critically appraising the existing evidence, we identify key knowledge gaps, such as the understudied sporadic EOPC subgroup and the lack of dedicated clinical trials, and propose future research directions to inform early detection and optimize therapeutic strategies for this unique patient population.

A recent individual patient data (IPD) meta-analysis of randomized trials found a limited overall survival (OS) and metastasis-free survival (MFS) benefit to the addition of hormonal therapy (HT) to postoperative radiotherapy for prostate cancer in patients with pre-radiotherapy prostate-specific antigen (PSA) ≤0.5 ng/ml. We evaluated whether the OS and MFS benefits of adding HT might be significantly modified by an increasing count of adverse pathological features (adverse feature count [AFC]). We obtained IPD from five randomized phase 3 trials of postoperative radiotherapy ± HT. AFC was prospectively defined as the sum (0-4) of grade group 4-5 disease, seminal vesicle invasion, positive surgical margins, and extracapsular extension. Intention-to-treat one-stage meta-analytical models were used. We also evaluated the interaction of the benefit of HT with a restricted score comprising only GG4-5 and seminal vesicle invasion (range 0-2), as a pre-specified sensitivity analysis. We included IPD from 4781 patients with a median follow-up of 9.1 yr. AFC was independently prognostic for OS and MFS. However, AFC did not significantly modify the OS benefit of HT (interaction hazard ratio [HR], 0.93; 95% confidence interval [CI], 0.79-1.10; p = 0.4) or MFS benefit (interaction HR, 0.88; 95% CI, 0.77-1.01; p = 0.08). Results were similar when AFC was analyzed categorically and in high-risk subsets, and when the restricted AFC score was used. Overall, the data suggest that although adverse pathological features are prognostic, they do not appear predictive of HT benefit after postoperative radiotherapy, which is relevant for patients with a PSA ≤0.5 ng/ml before radiation.

Immune checkpoint blockade is minimally active in unselected castration-resistant prostate cancer (CRPC) and does not reproducibly yield durable decreases in prostate-specific antigen (PSA) levels. The Quick Efficacy Seeking Trial was designed to employ a combination of agents to initiate an immune response (with BN-Brachyury vaccine), potentiate that response (with nogapendekin-alfa inbakicept (NAI), an interleukin (IL)-15 receptor superagonist), and reduce or eliminate immunosuppressive entities in the tumor microenvironment (with bintrafusp alfa, a dual inhibitor of programmed death-ligand 1 and transforming growth factor beta). Epacadostat (an indoleamine 2,3-dioxygenase (IDO) inhibitor) was also employed in one cohort to reduce immune suppression induced by IDO's conversion of tryptophan to kynurenine. Patients with CRPC enrolled sequentially to receive vaccine + bintrafusp alfa (Arm 2.1), vaccine + bintrafusp alfa + NAI (Arm 2.2), and vaccine + bintrafusp alfa + NAI + epacadostat (Arm 2.3), with the primary objective to determine response rate. Adverse events in Arms 2.1 and 2.2 were manageable and consistent with the safety profiles of each agent individually, and notable for five individuals developing isolated adrenocorticotropic hormone deficiency. Arm 2.3 was closed early due to skin toxicity. Sustained declines in PSA were seen in 1/13 (8%) patients in Arm 2.1, 7/24 (29%) patients in Arm 2.2, including six with proficient mismatch repair/microsatellite stable tumors, and 0/6 (0%) patients in Arm 2.3. Analyses of peripheral immune profiles provided evidence of a multifaceted antitumor immune response, including IL-15 receptor superagonist NAI-dependent expansion and activation of natural killer cells and CD8+ T cells, increased effector-to-suppressor immune cell ratios, and induction of cytotoxic immune gene programs. NCT03493945.

Upfront combination therapy in mHSPC improves outcomes; however, without guaranteed postprogression therapy in control arms, survival benefits may partly reflect undertreatment in the control groups rather than true superiority. Trial designs incorporating planned treatment sequencing are needed to support rational deescalation strategies.

Metastatic castration-resistant prostate cancer (mCRPC) is among the leading causes of cancer-related mortality in men worldwide. Treatment options for mCRPC typically include chemotherapy and androgen receptor pathway inhibitors. Immune checkpoint inhibitors (ICIs) have demonstrated a limited effect in mCRPC. We hypothesized that the addition of stereotactic body radiation therapy (SBRT) could enhance immune responses and improve treatment outcomes. Patients with mCRPC who had received at least two prior lines of therapy were randomized 1:1 to receive SBRT with nivolumab and ipilimumab (arm A) or nivolumab and ipilimumab (arm B). The dual primary endpoints in the study were prostate-specific antigen (PSA) response rate and objective response rate (ORR). Secondary endpoints included overall survival (OS), PSA, radiologic progression-free survival, and toxicity. We enrolled 91 patients, and 81 patients received at least one treatment with ICIs and were eligible for evaluation of the efficacy and safety endpoints. Of the evaluable patient population, the confirmed PSA response rate was 21.6% (95% CI 9.8% to 38.2%) in arm A, and 20.5% (95% CI 9.8% to 35.3%) in arm B. ORR was 16.7% (95% CI 4.7% to 37.4%) and 22.2% (95% CI 10.1% to 39.2%) in arms A and B, respectively. Median OS was 10.2 months (95% CI 7.1 to 15.2) in arm A and 9.2 months (95% CI 7.1 to 14.4) in arm B. Treatment-related adverse events grade 3-4 were observed in 27 patients (33.3%). The addition of SBRT to nivolumab and ipilimumab did not improve outcomes; however, a fraction of the patients had a response to the treatment combination of nivolumab and ipilimumab. Explorative translational research is needed to identify possible biomarkers of response to immunotherapy with ICIs in mCRPC. European Union Clinical Trial Registry (https://www.clinicaltrialsregister.eu) EudraCT number: 2018-003461-34 and on https://clinicaltrials.gov (NCT05655715).

AR pathway-independent prostate cancer (ARIPC), particularly neuroendocrine prostate cancer (NEPC), represents one of the most lethal states of metastatic castration-resistant prostate cancer. However, how fatty acid synthesis (FAS) is organized in ARIPC and whether distinct lipogenic states shape neuroendocrine lineage transdifferentiation remain unclear. By integrating single-cell and bulk transcriptomic analyses of mCRPC cohorts, we identify NEPC as a fatty-acid-synthesis-low state associated with poor survival. Within this context, fatty acid synthase (FASN) emerges as a key indicator and functional contributor to lipogenic activity. FASN depletion suppresses lipogenesis while increasing NEPC-associated programs, migration, and metastatic colonization. We further identify FGFRL1 as the FGF family member most consistently associated with fatty acid synthesis activity in ARIPC. FGFRL1 depletion reduces FASN expression and relative free fatty-acid content, while targeted GC-MS supports broader fatty-acid remodeling and fluorescent uptake assays show increased exogenous fatty-acid uptake. FASN restoration partially restores relative free fatty-acid content and attenuates NEPC-associated and migratory phenotypes. Directional perturbation, rescue, AKT phosphorylation, and co-immunoprecipitation analyses further support the functional FGFRL1-FASN relationship. ONECUT2 is prioritized as a candidate downstream transcriptional regulator whose expression correlates with the neuroendocrine program. Together, these findings support an FGFRL1-FASN metabolic axis that regulates neuroendocrine lineage transdifferentiation and metastatic progression in ARIPC.

Prostate cancer (PCa) is one of the most prevalent malignancies in men and frequently progresses to bone metastasis. Understanding the interactions between PCa cells and the bone microenvironment, as well as their impact on therapeutic response, is therefore of critical clinical relevance. This study aimed to investigate the effects of the osteogenic microenvironment on prostate cancer cells by constructing three-dimensional (3D) bioprinted in vitro co-culture models, and to evaluate how the osteogenic niche influences tumor malignant phenotypes. This study employed extrusion-based 3D bioprinting (3DP) to construct in vitro co-culture models of the PCa osteogenic microenvironment. Two 3D-PCa models based on LNCaP and PC-3 cells were each co-cultured with osteogenically differentiated adipose-derived stem cells (ADSCs) to generate corresponding osteogenic niche models. Tumor phenotypes and drug responses were evaluated using functional assays, histological and immunofluorescence analyses, molecular profiling, and RNA sequencing. The 3D bioprinted constructs exhibited structural stability and high reproducibility, providing a 3D growth environment that mimics key aspects of the in vivo tumor niche. The 3D-PCa models showed enhanced drug resistance, invasive potential, and adaptation to androgen-deprivation. Meanwhile, ADSCs exhibited robust osteogenic differentiation within 3D scaffolds. Under co-culture conditions, 3D-PCa cells exhibited enhanced malignant-like phenotypes, including increased proliferation and reduced drug sensitivity. Multi-level analyses further indicated that these phenotypic changes were associated with coordinated activation of epithelial-mesenchymal transition (EMT)-, hypoxia-, and mitogen-activated protein kinase (MAPK)-related signaling programs, accompanied by alterations in drug transporter expression and cell cycle distribution. The 3D co-culture model provides a biomimetic platform for simulating the PCa bone-metastatic microenvironment. This system enables investigation of the effects of the osteogenic niche on tumor behavior and therapeutic responses, and may serve as a useful tool for preclinical drug screening and evaluation of combination treatment strategies for bone-metastatic prostate cancer.

Ferroptosis is triggered by iron-dependent lipid peroxidation, which shows great potential in tumor therapy. However, its biomedical applications are constrained by suboptimal therapeutic efficacy, leakage of metal ions from inducing agents, and the absence of noninvasive evaluation methods. In this study, we aimed to develop a self-reducing nano-theranostic agent (Fe@AP-SAS/RGD, FASR) for tumor-targeted ferroptosis therapy and conducted real-time magnetic resonance imaging(MRI) to monitor ferroptosis by encapsulating iron oxide particles in natural apple pectin (AP). In a weakly acidic tumor microenvironment, the strong reducing ability of AP accelerated the transformation of Fe3+ within FASR into highly active Fe2+. Subsequently, the concentration of Fe2+ increased and boosted the antitumor effect of ferroptosis. Notably, MRI of prostate cancer mouse models after intravenous injection of FASR NPs at varying doses revealed that the T2 value decreased at the tumor sites and exhibited a linear correlation with the changes in ferroptosis hallmark markers, tumor weight, and volume after treatment. These findings demonstrate the capacity of T2 mapping to quantitatively evaluate ferroptosis efficacy. This study not only introduced a tumor-targeted and self-reducing strategy to amplify ferroptosis but also pioneered a novel pathway for noninvasive imaging and monitoring of ferroptosis treatment processes within live tumors.

High-grade central nervous system (CNS) tumors carry a poor prognosis with limited curative options if first-line therapy fails. B7-H3 is expressed in many of these tumors, and chimeric antigen receptor (CAR) T cell therapy is an emerging immunotherapeutic strategy. BrainChild-03 (NCT04185038) is a single-center, dose-escalation phase 1 study of repeated intracerebroventricular (ICV) B7-H3 CAR T cells in children and young adults with recurrent/refractory CNS tumors (Arms A, B) and diffuse intrinsic pontine glioma (DIPG, Arm C). Here, we report results from Arm B, in which patients with refractory/relapsed CNS tumors or pre- or post-progression non-pontine diffuse midline glioma (DMG) received repeated ICV infusions. Primary objectives were feasibility and safety/tolerability; secondary objectives included CAR T cell detection, disease response, and survival. Of 36 enrolled patients (atypical teratoid rhabdoid tumor n = 5, DMG n = 8, embryonal tumor with multilayer rosettes n = 2, ependymoma n = 4, high-grade glioma n = 6, medulloblastoma n = 8, pineoblastoma n = 3), manufacturing was successful for 35 patients, 26 of whom received therapy. Median age was 10 years (range 1-26). Dose escalation from 1 × 107 to 10 × 107 CAR T cells/dose identified this dose as the maximally tolerated dose regimen, with no dose-limiting toxicities observed. Across 181 total doses (median 7/patient), common adverse events included headache (n = 26), fever (n = 15), and nausea (n = 14). Median survival from first infusion was 11.5 months, ranging from 3.2 months (pineoblastoma, HGG) to 21.4 months (ependymoma); two patients achieved a partial response. Repeated ICV B7-H3 CAR T cell dosing is feasible and tolerable across a spectrum of pediatric CNS tumors, supporting continued investigation in future trials.

Diffuse midline glioma (DMG) is a highly aggressive pediatric brain tumor that is difficult to treat because the blood-brain-tumor barrier (BBTB) prevents most drugs from reaching the tumor. A major obstacle for understanding this tumor-vascular niche is the lack of a perfusable vascular model of DMG. To address this, we have established an in vitro model of the DMG-BBTB, comprised of patient-derived DMG tumor cells and primary human brain microvascular endothelial cells, astrocytes, and vascular pericytes, embedded in a fibrin matrix within a microfluidic chip. We report the development and characterization of self-assembled microvascular networks, with consistent vasculature development across a range of tumor cell densities. Perfusion studies showed no significant alterations in vascular permeability when tumor cells were present. To elucidate transcriptomic changes, we employed scRNAseq on devices containing blood-brain barrier (BBB), DMG-BBTB, or tumor necrosis factor alpha (TNFα)-stimulated control. We observed significant differential gene expression between BBB and DMG-BBTB, distinct from inflammatory signatures seen with TNFα perturbation. Our analysis indicated that DMG cells adopted a mesenchymal-like phenotype, with further in silico studies predicting significant receptor-ligand crosstalk between these cells and endothelium. In summary, we developed a high-fidelity DMG-BBTB model that recapitulates transcriptomic features of patient DMG microvasculature and features perfusable vasculature.

Glioblastomas function as intricate cellular networks that extend into the surrounding brain tissue, facilitating long-distance communication. This malignant connectivity spans from the tumor core to remote infiltration zones, in support of the concept of glioblastoma as a whole-brain disease. With growing ethical concerns in biomedical research and the inherent limitations of animal models in recapitulating human glioblastoma biology, there is an increasing demand for human ex vivo platforms capable of capturing the full infiltration spectrum from the tumor core to single-cell dispersion. Here we present a 3D, fully human ex vivo glioblastoma model (Core2Edge) that replicates this extensive infiltration range while preserving the intratumoral heterogeneity of the original tumor. This model involves implanting fluorescently labeled human glioblastoma organoids (GBOs) into organotypic human brain slices, maintaining the genetic integrity and cytoarchitecture of both brain and tumor. By combining tissue expansion with light-sheet fluorescence microscopy, we achieve high-resolution, 3D imaging of the entire GBO-brain slice model. This approach allows the study of initial infiltration steps, in-depth analysis of the invasive front, and exploration of cell-cell interactions between tumor cells and the tumor microenvironment, and offers a platform for drug screening and testing, reducing the need for animal models. Once GBOs are prepared, the protocol takes ~7-12 d. Key steps include brain slice preparation (~4-6 h, depending on quantity), 1 d for initial culture before GBO staining and transplantation, a variable culture period (≤10 d), and fixation (~8 h). The protocol requires experience with human brain slice and organoid culture.

The lack of brain penetrant and biologically stable positron emission tomography reporter systems hampers the development of neurological disease models and the monitoring of gene delivery because existing approaches depend on endogenous receptors that vary unpredictably in pathology. HaloTag, a fully exogenous protein label that forms rapid and irreversible bonds with synthetic ligands, provides a modular platform for engineering reporter probes with defined chemical properties. Here we developed a fluorine-18-labelled small-molecule HaloTag ligand optimized for brain entry and covalent retention at the reporter. The tracer showed specific binding in human cells expressing HaloTag and enabled non-invasive imaging of viral gene transfer to striatal neurons in mice, with clear detection of reporter expressing tissue and rapid clearance from surrounding regions. Optical imaging confirmed viral distribution and reporter expression, and a transgenic model expressing HaloTag fused to a postsynaptic protein demonstrated detection of physiologically expressed intraneuronal targets. This system establishes a modular platform for validating preclinical models and quantifying gene expression in the living brain.

Glioma is the most common malignant tumor of the central nervous system, with high malignancy and poor prognosis, necessitating the development of novel targeted therapies. DNA polymerase delta catalytic subunit 1 (POLD1) is implicated in multiple cancers, but its role in glioma remains unclear. Plant-derived extracellular vesicles (PDEVs) have emerged as biocompatible, targetable nanocarriers with promising applications in cancer therapy. This study aims to elucidate the oncogenic function of POLD1 in glioma and develop a PDEVs -based delivery system for targeted therapy, with the goal of improving the current therapeutic landscape for glioma. POLD1 expression and prognostic significance were analyzed using clinical samples and databases. In vitro, CCK-8, Transwell, and flow cytometry assays evaluated the impact of POLD1 knockdown on glioma cell proliferation, invasion, migration, cell cycle, and apoptosis. In vivo tumorigenesis and survival were assessed in mouse models. Sweet potato-derived nano-vesicles (SPDELNVs) were isolated and characterized. An engineered A2-SPDELNVs-siPOLD1 system was developed via surface modification and siRNA loading, and its targeting efficiency and therapeutic efficacy were evaluated both in vitro and in vivo. POLD1 was upregulated in glioma tissues and correlated with poor prognosis. Its knockdown suppressed proliferation, invasion, and migration, induced cell cycle arrest, and promoted apoptosis in vitro. In vivo, POLD1 targeting inhibited tumor growth and prolonged survival. SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake. The engineered A2-SPDELNVs-siPOLD1 effectively delivered siRNA, silenced POLD1, and significantly inhibited tumor progression both in vitro and in vivo, with enhanced survival. Our findings uncover the oncogenic role of POLD1 in glioma and validate it as a promising therapeutic target. Furthermore, we establish a novel, plant-based A2-SPDELNVs-siPOLD1 delivery platform with effective BBB penetration and tumor targeting, offering a promising strategy for the treatment of glioma.

Central nervous system (CNS) tumors are the most common solid malignancies in children, comprising a highly heterogeneous group of neoplasms defined by distinct molecular alterations and clinical behaviors. Advances in molecular genetics have underscored the relevance of specific signaling pathways in driving pediatric tumorigenesis, among which the fibroblast growth factors (FGFs) and their receptors (FGFRs) have emerged as critical regulators of CNS development and tumor biology. The strict regulation of this system is frequently disrupted in cancer contexts, leading to abnormal activation that promotes tumor growth, invasion, and therapy resistance. Recent genomic profiling studies confirm that FGFR gene aberrations (i.e., activating point mutations, amplifications, and fusions) occur in a non-negligible fraction of pediatric CNS tumors. Notably, FGFR alterations are found in approximately 9% of pediatric gliomas, representing a higher incidence compared to adult gliomas. Across a broader survey of pediatric solid and brain tumors, activating FGFR aberrations were measurable in approximately 3% of cases. This prevalence establishes the FGF/FGFR axis as a measurable therapeutic vulnerability in defined patient subsets. Therefore, a deeper understanding of the role of the FGF/FGFR system across different pediatric CNS tumor subtypes is essential for elucidating disease mechanisms and identifying novel therapeutic opportunities. This review outlines the characteristics of major pediatric CNS tumors and discusses the role and potential therapeutic targeting of the FGF/FGFR system.

Malignant gliomas are lethal brain tumors characterized by profound local immunosuppression and a radically remodeled myeloid landscape. Although these tumors mobilize resident microglia and infiltrating monocyte-derived macrophages, the mechanisms governing their phenotypic convergence and diversification remain elusive. Here, we integrated single-cell profiling and spatial transcriptomics of glioma-associated microglia in the GL261 model. We identified distinct microglial states that aligned with tumor architecture, most notably Cst7-expressing disease-associated microglia (DAMs) that aggregated at the tumor invasive margin and exhibited a conserved transcriptional signature shared across various central nervous system pathologies. Interferon-γ and toll-like receptor signaling sequentially tuned stage-specific DAM features, including transient MHC-II expression and sustained PD-L1 upregulation, thereby recalibrating the local immune equilibrium by reshaping bidirectional DAM-T cell interactions during glioma progression. Our findings highlight microglial state transitions as a stage-specific layer of immune regulation in glioma that shapes T cell fate and support targeting microglial plasticity to rebalance anti-tumor immunity.

Sonic hedgehog-medulloblastoma (SHH-MB) represents a biologically diverse and clinically challenging subgroup, especially in high-risk variants characterized by metastatic dissemination. Despite progress in molecular stratification, current treatments rely on intensive multimodal regimens associated with substantial long-term neurotoxicity. Identifying novel oncogenic mechanisms that can be therapeutically exploited is therefore a critical priority. We combined transcriptomic analyses of patient datasets with immunohistochemistry, RNAscope, and protein profiling to evaluate PTX3 expression in MB subgroups. Functional studies were performed in SHH-MB cell lines following PTX3 knockdown or knockout. Effects on proliferation, migration, and angiogenesis were assessed through multiple in vitro assays. In vivo relevance was tested using subcutaneous and orthotopic xenograft models. Mechanistic insights were obtained through phospho-kinase arrays, Western blotting and GSEA. A high-throughput drug screen and combination studies with specific inhibitors and standard chemotherapy (vincristine/cisplatin/cyclophosphamide; VCC) were performed to assess therapeutic potential. Our results reveal that PTX3 was strongly enriched in SHH-MB subgroup, where it supported proliferation, motility, and angiogenesis. Mechanistically, PTX3 activated a TLR4-dependent IRAK1/PI3K-Akt/GSK-3/β-catenin signaling cascade. Genetic downregulation or pharmacologic blockade of PTX3-TLR4 significantly reduced tumor growth and angiogenesis in vivo. Moreover, the TLR4 inhibitor TAK-242 markedly impacted SHH-MB growth in vitro, and combination therapy of TAK-242 and VCC produced additive/synergistic effects in vitro and significantly prolonged survival in orthotopic SHH-MB-bearing mice. Our findings identify PTX3-TLR4 signaling as key oncogenic driver and a promising therapeutic vulnerability in SHH-MB. Targeting this pathway enhances the efficacy of standard therapy and represents a rationale for mechanism-based combination strategies.

Non-pharmacological treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) aims at modifying lifestyle patterns that promote metabolic dysfunction and hepatic fat accumulation. These include unhealthy dietary habits, which are characterised by excess energy intake and a qualitatively poor diet that is high in ultra-processed food, saturated fat, free sugar, red and processed meat and/or sweetened beverages, and low in fibre. In addition, sedentary habits, and sleep and circadian rhythm disruption have been identified as detrimental lifestyle factors. Lifestyle interventions act upstream of the main pathophysiological mechanisms driving MASLD and, therefore, provide benefits that extend beyond the liver, also improving cardiometabolic health. Weight loss remains the cornerstone of non-pharmacological strategies, and even modest weight loss has been proven beneficial in individuals with MASLD with or without obesity. To achieve weight loss, energy restriction should be combined with physical activity/exercise, since the effects of the latter are partly independent of weight reduction. Importantly, exercise is associated with a dose-dependent reduction in the risk of hepatic steatosis, fibrosis and hepatocellular carcinoma. Despite lifestyle changes proving highly effective for improving MASLD, their implementation is challenging and requires multidisciplinary efforts. Adherence is suboptimal, particularly in the long term, and the variability in individual response calls for research on biomarkers to achieve a more personalised approach. This review provides an overview of evidence of the impact of lifestyle changes/non-pharmacological interventions on MASLD.

Surgical resection is a cornerstone of curative therapy for hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (iCCA), the two most common primary liver malignancies. In most patients with HCC, underlying cirrhosis is present, whereas in iCCA an identifiable risk factor is often absent. Regardless of tumour type, surgical planning must account for the future liver remnant to minimise the risk of post-hepatectomy liver failure, evaluating not only its volume but also its functional reserve. Furthermore, a deeper knowledge of surgical oncological principles combined with integrative strategies, including systemic and locoregional therapies, has enabled downstaging and conversion of initially unresectable liver malignancies, expanding surgical eligibility across a growing spectrum of hepatic tumoural conditions. Additionally, advances in the understanding of molecular biology and new treatment strategies, particularly in iCCA, have the potential to further extend the boundaries of resectability.

Artificial intelligence (AI), particularly foundation and generative models, is reshaping the practice of hepatology through enhanced knowledge synthesis, quantitative and reproducible analysis of multimodal data, and personalized clinical decision support. This narrative review examines the transition from task-specific discrimination AI to large language models (LLMs), multimodal foundation models, and agentic AI. We synthesize evidence from original and validation studies, clinical evaluations, and benchmark studies, as well as expert reviews and regulatory frameworks across metabolic dysfunction-associated steatotic liver disease, chronic hepatitis B, cirrhosis and portal hypertension, hepatocellular carcinoma, and liver transplantation. LLMs can convert free-text notes into structured data, summarize longitudinal electronic health records, support patient education, and retrieve guideline-based information. Retrieval-augmented generation and agentic AI may improve traceability and workflow support, but current evidence is largely retrospective or proof-of-concept. In digital pathology and imaging, discriminative AI has enabled more quantitative and reproducible histologic scoring and biomarker analysis. Pathology and multimodal foundation models offer transferable representations, report generation, and cross-modal reasoning, but hepatology-specific validation remains limited. Key risks include hallucination, automation bias, domain shift across centers and devices, and inequities due to under-representation of patient subgroups. We outline the future directions for safe AI model deployment based on multimodal foundation models, prospective and federated evaluation, lifecycle governance, and continuous monitoring for performance, calibration, and equity. Most generative AI applications in hepatology remain at the proof-of-concept stage, and rigorous prospective validation with human-in-the-loop oversight is required before clinical integration.

The liver naturally exhibits robust glycosylation processes, yet how aberrant glycosylation affects hepatocellular carcinoma (HCC) and its underlying mechanisms remains poorly understood. To investigate the role of heparan sulphate (HS) modification in HCC progression and identify its underlying regulatory mechanism. The association of HS with poor prognosis and immune dysfunction was analysed through HCC clinical data and in vivo models. T cell functions were validated in vivo and in vitro via exogenous HS or HS depletion. Transcriptomic screening of HS-high and HS-low tumour clones was performed to identify upstream regulators of HS. Candidate galectin-14 was functionally confirmed through genetic manipulation in vitro and in transgenic mice. Metabolic and protein-interaction studies showed how galectin-14 drives UDP-glycosyl donor production and HS synthesis. Elevated HS modification correlated with poor survival and was linked to T cell exhaustion in patients with HCC. The primate-specific oncofetal protein galectin-14 was identified as an upstream regulator of HS synthesis. Galectin-14 promoted HCC progression and immune evasion by inducing contact-dependent CD8+ T cell exhaustion, partly mediated by FGFR1 signalling on T cells. It reprogrammed glycometabolism to boost synthesis of UDP-glycosyl donors (UDP-Glc, UDP-GlcNAc, UDP-GlcA) required for HS chains. Mechanistically, galectin-14 directly bound UDP-glucose pyrophosphorylase 2 (UGP2), facilitating its oligomerisation and enzymatic activity, thereby fueling HS production. This study defines galectin-14 as a key oncofetal regulator of HCC immunosuppression by rewiring UGP2-mediated glycosyl donor production to promote elevated HS modification and T cell dysfunction, highlighting a potential diagnostic marker and promising therapeutic avenue in HCC.

Hepatocellular carcinoma (HCC) is a leading cause of global cancer mortality, arising predominantly in the context of chronic liver injury and cirrhosis. The tumor microenvironment (TME) is a critical orchestrator of HCC progression and therapeutic resistance. Neutrophils, the most abundant circulating leukocytes and key first responders in hepatic immunity, play a pivotal and multifaceted role in this process. Under physiological conditions, neutrophils are essential for liver homeostasis and acute injury resolution. However, across the pathological continuum from chronic liver disease to HCC, sustained inflammatory signaling drives a profound reprogramming of neutrophil function. Within the established HCC TME, tumor-associated neutrophils (TANs) are frequently polarized into a protumorigenic phenotype, actively driving disease progression through several core mechanisms: they establish immunosuppressive niches by directly suppressing cytotoxic T-cell function; promote angiogenesis and metastasis via the secretion of factors like VEGF and MMPs; and contribute to genomic instability and therapy resistance through mechanisms such as neutrophil extracellular trap (NET) formation. This functional transition from defenders in acute injury to accomplices in malignant progression underscores the central role of neutrophils in liver pathobiology and HCC pathogenesis. This review synthesizes current knowledge on the recruitment, polarization, and functional diversity of neutrophils in the HCC TME, with a particular focus on their mechanisms in mediating immunosuppression through cellular crosstalk and metabolic reprogramming. We further critically evaluate emerging therapeutic strategies that target neutrophil-centric pathways to resuscitate antitumor immunity, including TAN repolarization, depletion of specific subsets, or functional blockade. Finally, we discuss the translational potential of neutrophil-related biomarkers for patient stratification and the rationale for developing combination therapies. A deeper understanding of the dynamic and pleiotropic roles of neutrophils in HCC is essential for devising more effective immunotherapeutic strategies and improving patient outcomes.

Inducing ferroptosis in hepatocellular carcinoma (HCC) cells represents an important therapeutic strategy, but intrinsic resistance mechanisms often limit efficacy. Therefore, elucidating the mechanisms underlying ferroptosis resistance in HCC cells can facilitate the development of effective therapeutic strategies. Here, we performed genome-wide CRISPR/Cas9 library screens to identify TRIM27 as a key determinant of ferroptosis resistance. TRIM27 knockdown markedly potentiated erastin-induced ferroptosis in HCC cells, whereas TRIM27 overexpression suppressed the expression of fatty-acid metabolic enzymes including ACSL4 and reduced oxidized lipid accumulation. Mechanistically, TRIM27 directly binds with ACSL4 and promotes its K48-linked ubiquitination and degradation, thereby attenuating ferroptosis in HCC cells. Furthermore, we developed TRIM27-Cas9-loaded EVs with robust editing efficiency. These engineered EVs were readily internalized by HCC cells and preferentially accumulated in the liver. Functionally, TRIM27-Cas9-loaded EVs inhibited HCC cell proliferation by enhancing ACSL4-mediated ferroptosis and significantly improved the anti-tumor efficacy of anti-PD-1 therapy in HCC. Collectively, our findings suggest that TRIM27 confers ferroptosis resistance via facilitating K48-linked ubiquitination and subsequent proteasomal degradation of ACSL4. TRIM27-Cas9-loaded EVs restore cellular sensitivity to ferroptosis, inhibit HCC proliferation, and sensitize HCC lesions to anti-PD-1 immunotherapy.

Mitophagy is a mitochondrial quality control process that maintains cellular homeostasis in cancer, yet whether its dysregulation can be exploited to induce tumor immunogenicity remains unclear. We integrated pancancer single-cell transcriptomic analyses with genetic perturbation strategies in hepatocellular carcinoma models, including CRISPR/Cas9-mediated gene depletion, in vivo syngeneic tumor systems, and RNA-based lipid nanoparticle delivery. Mechanistic investigations combined mitochondrial functional assays, imaging-based mitophagy analysis, flow cytometry, and transcriptional profiling, together with evaluation of immune checkpoint blockade responses in preclinical and clinical cohorts. We identify translocase of the outer mitochondrial membrane 40 (TOMM40) as a mitochondrial import gatekeeper that restrains PINK1-Parkin-dependent mitophagy. Loss of TOMM40 induces catastrophic mitochondrial dysfunction and triggers a lethal form of hyperactivated mitophagy. This process is immunogenic and converts immune-cold tumors into immune-inflamed states characterized by enhanced CD8+ T-cell infiltration and activation. Mechanistically, TOMM40 deficiency leads to intracellular reactive oxygen species accumulation, which activates NF-κB signaling and drives upregulation of major histocompatibility complex class I antigen presentation machinery, thereby increasing tumor visibility to cytotoxic T cells. In parallel, TOMM40 loss induces programmed death-ligand 1 upregulation, establishing an adaptive immune resistance program. Functionally, TOMM40-deficient tumors exhibit markedly increased responsiveness to immune checkpoint blockade and generate systemic antitumor immune protection. Clinically, a TOMM40-loss transcriptional signature is associated with improved immunotherapy outcomes across multiple independent patient cohorts. TOMM40 functions as a mitochondrial immune checkpoint that controls the threshold of immunogenic mitophagy. Its loss reprograms mitochondrial stress into antigen presentation and immune activation, providing a strategy to convert immune-cold tumors into immune-responsive states.

Photodynamic therapy (PDT) faces significant challenges in treating solid tumors due to the hypoxic tumor microenvironment and high degree of tumor heterogeneity. To address this issue, this study employed a strategy of acceptor planarization coupled with tunable terminal aryl modulation to design and synthesize a series of type I photosensitizers (DPP-1-DPP-3) with systematically tuned push-pull character and electronic structures. Among them, DPP-3 exhibits outstanding near-infrared emission and hypoxia-tolerant reactive oxygen species generation. Building on this core module, we integrated a biomarker-responsive unit into DPP-3 to construct an intelligent theranostic probe, DPP-CE. This probe retains high photodynamic activity both before and after activation, ensuring reliable therapeutic efficacy independent of local activation efficiency, while its near-infrared fluorescence signal is specifically activated only within the tumor microenvironment, enabling imaging-guided precise treatment. Both in vitro and in vivo experiments demonstrate that DPP-CE allows high-contrast fluorescence imaging of tumors and effectively inhibits tumor growth under both normoxic and hypoxic conditions, while showing good biosafety. This work not only provides a new strategy for developing high-performance type I photosensitizers, but also offers a modular approach that integrates a therapeutic core with a biomarker-responsive unit, paving the way toward programmable theranostic platforms adaptable to tumor heterogeneity.

Chemotherapy can induce tumor cell death and stimulate immunity, yet its efficacy is often compromised by the immunosuppressive tumor microenvironment dominated by M2-like tumor-associated macrophages (TAMs). Redirecting TAMs toward an M1-like phenotype through mammalian target of rapamycin (mTOR)-mediated metabolic modulation has therefore emerged as a promising therapeutic strategy. To this end, we developed a carrier-free nanoassembly (SOP) composed of the mTOR inhibitor OSI-027 and the chemotherapeutic agent SN-38. SOP harnesses ribosome-inspired crowding control to suppress liquid-liquid phase separation (LLPS), thereby facilitating M2 reprogramming and enhancing antitumor efficacy. Following intravenous administration, SOP accumulated in tumor tissues with 2-fold greater efficiency than free drugs. Once internalized, SOP released SN-38 to induce tumor cell death and immunogenic responses, while OSI-027 simultaneously inhibited mTORC1/2 signaling and reduced ribosome abundance-key crowding agents driving LLPS. The resulting disruption of LLPS promoted TAM repolarization toward M1, establishing a synergistic interplay between OSI-027 and SN-38. This dual action translated into robust therapeutic outcomes across diverse models, including cell-derived xenografts, patient-derived xenografts, and KPC (LSL-KrasG12D/+; LSL-Trp53R172H/+; Pdx-1-Cre) mice. Moreover, SOP suppressed resistance-related gene expression, overcoming chemotherapy resistance. In summary, this carrier-free nanoassembly not only reprograms the immunosuppressive tumor microenvironment but also introduces LLPS suppression as a mechanism for TAM regulation, positioning SOP as a potent platform for enhanced chemo-immunotherapy.

Mucosal immunity-the body's frontline defense, harboring 80% of the body's immune cells-represents a potent yet underexploited avenue for cancer vaccination. Here, we developed an oral biohybrid vaccine platform by integrating tumor antigen-loaded liposomes with fimbriae-enriched bacteria (Escherichia coli or VNP20009) through bacterial hitchhiking or membrane hybridization. These biohybrids promote mucosal antigen delivery via glycoprotein 2 (GP2)-mediated microfold-cell (M-cell) transcytosis, enhancing antigen cross-presentation and activation of a mucosa-periphery-tumor immune cascade. Bacterial membrane-hybridized vaccines outperform bacteria-hitchhiking counterparts by reconfiguring dendritic cell (DC) subsets within gut-associated lymphoid tissues (GALTs) and triggering C-C chemokine receptor type 7 (CCR7)-dependent immune cell trafficking, thereby propagating mucosal immune activation toward distal tumor microenvironment (TME) reprogramming and tumor control. When combined with PD-1 blockade, this strategy enhances antitumor efficacy by promoting effector cell mobilization and establishing memory against tumor rechallenge. Collectively, these findings position bacteria-derived arsenal biohybrids as a versatile oral vaccine strategy, advancing mucosal immunotherapy for cancer.

Ultrasound is the second most common clinical imaging modality. Ultrasound image quality often suffers from poor contrast resolution and thus can greatly benefit from appropriate contrast agents. While micron-sized gas core particles (microbubbles) are clinically utilized in this space for applications such as echocardiography, a smaller agent could be more broadly applicable to enhance the sensitivity and specificity of disease detection. To this end, lipid-shelled nanobubbles have recently emerged as robust ultrasound contrast agents for both diagnostic and therapeutic purposes. They have been used in preclinical applications ranging from oncology to endocrinology and are notably relevant in diseases that involve pathological vasculature. Owing to their shell composition, nanobubbles are versatile and can be functionalized with fluorophores, targeting agents and therapeutic moieties. Here we provide the steps needed to formulate plain, fluorophore-conjugated, ligand-conjugated, hydrophilic dye-integrated and drug-loaded nanobubbles. The typical formulation for lipid-shelled nanobubbles takes place via self-assembly driven by mechanical agitation, followed by differential centrifugation. The process is deceptively simple, yet there are many nuances in the formulation process that must be followed to produce consistently successful nanobubble batches without contamination from microbubbles. Even minor deviation from the standard protocol can substantially affect nanobubble yield, stability, acoustic performance and batch-to-batch reproducibility. This procedure also details common pitfalls and their potential consequences for bubble quality and performance. The procedure requires 3 h to complete the formulation and activation of the plain nanobubbles by users with basic laboratory expertise.

Horizontal mitochondrial transfer (HMT) outcomes are shaped by donor fitness and transfer context. We propose a post-transfer quality checkpoint that integrates membrane potential, oxidative damage, mitophagy, fusion, and fission to determine the recipient-cell's response. Depending on donor quality and recipient thresholds, HMT may drive bioenergetic restoration, inflammation, or tumor immune escape. This framework extends route-centered accounts of HMT toward a testable, quality-governed model for therapeutic intervention.

Conventional cyanine photosensitizers face challenges in photodynamic therapy (PDT) due to poor intersystem crossing (ISC), hypoxia sensitivity, and low tumor retention. Herein, we report the first asymmetric D-π-A-D' cyanine dyes with hybridized local and charge-transfer (HLCT) characteristics to address these limitations. By precisely regulating triplet excitons of the T2 state, these dyes facilitate enhanced ISC from the S1 to T2 state and boost reactive oxygen species (ROS) generation. These HLCT-type cyanine dyes integrate the charge-transfer state to promote oxygen-independent Type I PDT for hypoxic tumor environments and the locally excited state to maintain high photoluminescence quantum yield (PLQY) for visualized tumor therapy. Among them, CyTY-3 exhibits exceptional dual Type I/II PDT performance, with a singlet oxygen yield outperforming ICG by 27.8-fold, while simultaneously delivering a high NIR-II PLQY of 11.4% for high-contrast imaging. Unique delocalized charge distribution enables excellent tumor targeting and an ultralong retention time of up to 12 days. In vivo, CyTY-3 nanoparticles achieve 99.01% tumor inhibition under 808 nm irradiation with good biosafety. This HLCT strategy provides a robust platform for developing next-generation photosensitizers to overcome the intrinsic limitations of hypoxic tumor therapy.

The persistent issue of poor prognosis in tumor recurrence and metastasis, even with promising treatments such as photodynamic therapy, underscores the urgent need for noninvasive imaging tools to investigate underlying mechanisms. However, achieving real-time, crosstalk-free monitoring of multiple biomarkers within pathological processes remains challenging, largely due to the absence of effective molecular design strategies. Herein, we propose a "spatiotemporal asynchronous" probe design strategy and develop HD-LN, a sequentially activatable fluorescence/photoacoustic (FL/PA) probe for real-time monitoring of neutrophil recruitment after tumor phototherapy. HD-LN is first activated by tumor-overexpressed leucine aminopeptidase (LAP), leading to fluorescence activation and restoration of photodynamic (PDT) activity. After phototherapy, it is further activated by recruited neutrophil elastase (NE), resulting in fluorescence quenching together with enhanced photoacoustic signaling. Through this sequential activation of FL/PA signals, we successfully demonstrated that poor PDT prognosis correlates with neutrophil recruitment and NETs-associated changes during treatment, while inhibition of NE expression significantly improves therapeutic outcomes and prevents tumor metastasis. Furthermore, by leveraging the sequential signal changes, HD-LN first enabled in vivo screening of NE inhibitors. This work not only provides insights into overcoming poor PDT prognosis but also establishes a novel strategy for real-time, crosstalk-free monitoring of multiple biomarkers during physiological processes.

Clinical complete response to systemic therapy for patients with muscle-invasive bladder cancer should be approached as rigorously as one would approach any biomarker. This requires uniform assessments and consistent definitions. Emerging data indicate that clinical response assessment may allow individualized treatment for patients with muscle-invasive bladder cancer but further studies are needed.

Platinum-based chemotherapy (PBC) is frequently used after first-line enfortumab vedotin-pembrolizumab for metastatic urothelial carcinoma (mUC), yet efficacy in this setting remains undefined. We retrospectively reviewed an institutional database of 236 patients with mUC treated with enfortumab vedotin-pembrolizumab between October 2018 and December 2024 to identify those receiving second-line PBC. Responses were investigator-assessed. Progression-free survival (PFS) and overall survival (OS) were estimated from the start of PBC; duration of response (DOR) was estimated from the first documented response. Of 62 patients treated with subsequent systemic therapy, 74% (n = 46) received PBC (carboplatin, n = 34; cisplatin, n = 12); nine patients (20%) then received maintenance avelumab. Median age was 73 years; 65% were male; 30% had upper-tract primary. Objective response rate (ORR) to first-line enfortumab vedotin-pembrolizumab was 41%. Median follow-up was 11 months [95% confidence interval (CI) 9.2-not reached (NR)]. ORR to second-line PBC was 49% (22 of 45, 95% CI 34% to 64%), including two complete responses; median DOR was 5.2 months (95% CI 3.8-NR). Median PFS was 5 months (95% CI 3.6-6.6), and median OS was 12 months (95% CI 10-17). Outcomes did not differ between cisplatin and carboplatin (P > 0.9). In a real-world setting, PBC after enfortumab vedotin-pembrolizumab demonstrated meaningful activity but modest durability. These findings may inform clinical management and future trial design.

The perioperative treatment landscape of muscle-invasive bladder cancer (MIBC) is rapidly evolving with the introduction of immune checkpoint inhibitor (ICI)- and antibody-drug conjugate (ADC)-based strategies. However, direct randomized comparisons among contemporary regimens are lacking, limiting the interpretation and positioning of treatments. We aimed to compare the efficacy of perioperative systemic treatment strategies for MIBC with a network meta-analysis (NMA) of randomized controlled trials (RCTs). PubMed, Embase, Web of Science, and congress abstracts were systematically searched up to 28 February 2026. Phase II-III RCTs enrolling patients with MIBC treated with perioperative systemic therapy initiated before radical cystectomy and reporting overall survival (OS), event-free survival (EFS), and/or pathological complete response (pCR) were included. A frequentist graph-theoretical NMA was carried out to compare three contemporary RCTs using a star-shaped network anchored to gemcitabine-cisplatin (GC) as the common comparator. Treatment ranking was estimated using P-scores. The primary outcome was OS. Secondary outcomes included EFS and pCR. In total, 2293 patients from three RCTs contributed to the primary network. Compared with GC, enfortumab vedotin-pembrolizumab (EVP) was associated with improved OS [hazard ratio (HR) 0.65], as was GC plus durvalumab (HR 0.75). EVP and GC plus durvalumab were also associated with improved EFS (EVP HR 0.53, GC plus durvalumab HR 0.68) and higher pCR rates [EVP odds ratio (OR) 2.62, GC plus durvalumab OR 1.57] compared with GC. Across evaluated endpoints, EVP consistently achieved the most relevant P-score-based ranking. Subgroup-specific and sensitivity analyses demonstrated a consistent treatment direction. In this NMA of RCTs, perioperative regimens incorporating ICIs and ADCs were associated with improved survival and pathologic response outcomes compared with GC alone. Given the sparse star-shaped network and the absence of closed loops, these findings should be interpreted as indirect comparative estimates rather than definitive evidence of treatment superiority, while still providing a clinically relevant framework for contextualizing contemporary perioperative regimens.

Bladder cancer remains a significant clinical challenge due to high recurrence and progression rates, necessitating novel therapeutic strategies. Oncolytic viruses, such as the herpes simplex virus type 2-based OHSV2, have demonstrated promising antitumor effects through direct oncolysis and immune activation. This study investigated the efficacy, safety, and molecular mechanisms of OHSV2 in the treatment of bladder cancer. In vitro and in vivo experiments demonstrated that OHSV2 potently inhibited bladder cancer cell proliferation, migration, and clonogenicity in a dose-dependent manner, while exhibiting a favorable safety profile. Critically, OHSV2 treatment triggered a pro-inflammatory tumor immune microenvironment, characterized by increased infiltration and activation of CD8+ T cells. Mechanistically, OHSV2 induced pyroptosis in bladder cancer cells via the canonical Caspase-1/Gasdermin D (GSDMD) pathway, accompanied by increased interleukin-18 (IL-18), interleukin-1β (IL-1β), and lactate dehydrogenase (LDH) release. Further analysis identified NOD-like receptor family pyrin domain containing 3 (NLRP3) as the key upstream pattern recognition receptor for Caspase-1/GSDMD activation, and Toll-like receptor 4 (TLR4) as a critical mediator of NLRP3-dependent pyroptosis. Inhibition of TLR4 or NLRP3 partially reversed OHSV2-induced cytotoxicity, confirming their functional roles. Additionally, combining OHSV2 with the TLR4 agonist enhanced pyroptosis in a subcutaneous xenograft model, amplified antitumor immunity, and improved tumor control in vivo, suggesting potential synergism for clinical translation. These findings elucidate a novel TLR4/NLRP3/Caspase-1/GSDMD axis as the core mechanism behind the antitumor effect of OHSV2 and propose a rationale for its combination with immunomodulators to improve outcomes in bladder cancer.

Ferroptosis is an iron-dependent form of regulated cell death driven by phospholipid peroxidation, and it has emerged as a potential therapeutic vulnerability of cancer. Here we identify the secretory phospholipase PLA2G2F (phospholipase A2 group IIF) as a ferroptosis suppressor in bladder cancer and elucidate its regulation and mechanism of action. PLA2G2F functions through an intracellular mechanism by localizing to the endoplasmic reticulum to inhibit ferroptosis. Our genetic and pharmacological analyses reveal that peroxisome proliferator-activated receptor γ (PPARG), a nuclear hormone receptor and transcription factor previously implicated in ferroptosis regulation, upregulates PLA2G2F and that PPARG-mediated ferroptosis resistance is largely dependent on PLA2G2F in bladder cancer. Further, lipidomic profiling suggests that PLA2G2F preferentially acts on ether-linked phospholipids containing polyunsaturated fatty acids, thereby reducing the pool of peroxidation-prone polyunsaturated fatty acid-containing phospholipids. Together, our findings establish PLA2G2F as an endoplasmic reticulum-resident ferroptosis suppressor regulated by PPARG and show that inhibiting PPARG signaling or PLA2G2F activity can sensitize bladder cancer cells to ferroptosis induction.

The intratumoral microbiome has recently emerged as a potential hallmark of cancer, with implications for response or resistance to therapy. Bacteria can either promote or inhibit cancer growth. However, intratumoral bacteria can also be engineered using synthetic biology to remodel the tumor microenvironment. Here, we engineered the probiotic bacterium Escherichia coli Nissle 1917 (EcN) to express the human chemokine CXCL13 (C-X-C motif chemokine ligand 13), a critical component of germinal center (GC) formation. Antibody affinity maturation and class switching are fundamental aspects of adaptive immune response. Both occur primarily in the GCs of secondary lymphoid organs for defense against pathogens. Immune checkpoint blockade (ICB) efficacy is primarily driven by T cells; however, recent studies in mice and humans have shown that humoral immune responses act as critical partners for ICB-mediated antitumor activity. Using orthotopic models of bladder cancer, intravesically delivered engineered CXCL13-expressing EcN colonized bladder tumors and elicited GC responses in bladder tumor-draining lymph nodes after intravesical delivery. When combined with programmed cell death protein 1 (PD-1) blockade, engineered EcN improved antitumor activity in two aggressive, fast-growing, and immunologically cold orthotopic mouse models of bladder cancer. Mechanistically, this antitumor effect was dependent on the presence of CD8+ T cells and CD4+ T follicular helper cells; combination therapy increased tumor-specific antibody responses and promoted long-term survival and protective immunity upon tumor rechallenge. Thus, we demonstrate that synthetically engineered CXCL13-expressing EcN can enhance the efficacy of PD-1 checkpoint blockade immunotherapy by amplifying tumor-specific humoral immunity.

The efficacy of postsurgical immunotherapy for gastric cancer (GC) remains controversial, and the effect of the number of dissected lymph nodes (DLNs) on the efficacy of immunotherapy remains unknown. We retrospectively reviewed 451 patients who received adjuvant or palliative therapies (including immunotherapy and chemotherapy) for GC at nationwide centers between 2020 and 2025 to investigate the influence of DLNs on postsurgical immunotherapy. Prognoses were assessed based on DLNs. In the adjuvant immunotherapy cohort (n = 139), the disease-free survival (DFS) was significantly lower in patients with DLNs >36 compared with those with DLNs ≤36 [hazard ratio (HR), 2.62; 95% confidence interval (CI), 1.39-4.95, P = 0.003)]. Moreover, patients with DLNs≤36 who received combined immunotherapy had a better DFS than patients who received only adjuvant chemotherapy (HR, 0.37; 95% CI, 0.17-0.78, P = 0.006). In the palliative immunotherapy cohort (n = 47), patients with DLNs>36 had significantly lower progression-free survival (PFS) (HR, 3.57; 95% CI, 1.61-7.94, P = 0.001) and DCR (6.67% vs 65.6%, P < 0.001) than those with DLNs≤36. Additionally, when matching with patients who received palliative immunotherapy without previous gastrectomy (DLNs = 0), the PFS decreased progressively in patients with DLNs = 0, DLNs>0 to ≤36, and DLNs>36 (median PFS, 16 vs. 11 vs. 3 months, P = 0.002). In conclusion, excessive lymph node dissection (DLNs≥36) compromised the efficacy of postsurgical adjuvant or palliative immunotherapy for GC.

Extracellular vesicles (EVs) are promising liquid biopsy candidates for gastric cancer because they can capture tumor-, immune-, microbial-, and microenvironment-related signals. We argue that the main challenge is no longer simply to find more candidates but to advance EV biomarkers with demonstrated diagnostic, therapeutic-monitoring, or prognostic value toward clinical translation. In this opinion article, we highlight EVs from nonblood biofluids, non-tumor-derived EVs, and underexplored EV cargo types, particularly small noncoding RNA cargoes, as sources of biomarker potential. We further identify insufficient clinical evidence, technical bottlenecks, and incomplete quality-control and regulatory standards as key barriers to their clinical deployment.

Helicobacter pylori-associated stomach adenocarcinoma (STAD) represents a highly severe malady, with up to 1 million new cases annually. Here, we examined novel human and bacterial risk determinants as well as related signal transduction events associated with STAD development. H. pylori serine protease HtrA cleaves the junctional protein E-cadherin, which results in the disruption of epithelial cell connections, the release and nuclear accumulation of β-catenin, and the onset of epithelial-mesenchymal transition (EMT), a hallmark of many tumors. In addition, the injection of H. pylori oncoprotein CagA into host epithelial cells targets β-catenin-mediated cell proliferation and other cancer signaling pathways. By analyzing over 2,000 H. pylori genomes, we identified single-nucleotide polymorphism (SNP) variants of HtrA and CagA that are associated with STAD progression. In addition, we investigated the role of the human serine proteases HTRA1, HTRA2, HTRA3, and HTRA4 in STAD progression and linked the genetic and expression data with specific signaling pathways. Elevated HTRA1, HTRA2, and HTRA3 expression in STAD patients correlated with upregulated extracellular matrix (ECM) receptor interactions and signaling that are critical for EMT. Moreover, H. pylori-positive STAD patients exhibited increased epithelial cell signaling, chronic inflammation, transcription factor Wnt/β-catenin signaling, ECM damage and metastasis, and single-cell analyses showed a strong association between HTRA1, the receptor Wnt, β-catenin, and oncogene MYC expression. Analyses of mutations in human HTRA1 and H. pylori HtrA revealed a role in the up- or downregulation of STAD progression. Together, our data show that SNPs in human and H. pylori serine protease HtrA and CagA modulate cancer signaling in complex Wnt-/β-catenin and ECM signaling networks, and that the protein variants can be causative or protective factors. A signaling model is proposed that highlights the complex interplay of human and bacterial factors in critical tumor signaling events, which could serve as predictive STAD biomarkers in patients.

Patients with gastric cancer (GC) and peritoneal metastasis (PM) have poor prognoses due to drug resistance and metastatic relapse. The mechanism underlying PM recurrence remains unclear. This study investigates the mechanisms of ascitic neutrophil extracellular traps (NETs) in promoting GC immune escape and PM recurrence. Ascites from a patient with GC were analysed using flow cytometry, immunofluorescence and electron microscopy. In vitro and in vivo experiments demonstrated NETs promoted GC immune evasion. RNA sequencing and functional models identified the immune checkpoint molecule CEACAM1. Internalisation assay combined with inhibitors determined tumour cells internalised NETs via macropinocytosis. Immunofluorescence and western blot revealed that NETs upregulated CEACAM1 and programmed death-ligand 1 (PD-L1) by activating the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway. The cotranscription factor interferon regulatory factor 1(IRF1) for CEACAM1 and PD-L1 were identified by dual-luciferase reporter assays and chromatin immunoprecipitation. Conditioned medium-culture systems identified CSF2 and CSF3 as NETs-inducing factors. Combination of DNase I and PD-L1 antibody was evaluated in humanised mice, organoid-T cell cocultures and ex-vivo ascites therapy models. We uncovered that macropinocytosis mediated uptake of NETs remodels an immunosuppressive microenvironment and promotes PM recurrence. Increased neutrophils and NETs in PM-relapsed ascites promote GC cell immune escape. Mechanistically, GC cells uptake NETs-DNA via macropinocytosis. Macropinocytic NETs-DNA activate the cGAS-STING-IRF1 pathway, upregulate immune-checkpoint molecules CEACAM1 and PD-L1, leading to immune escape. Moreover, clearing NETs-DNA combining with PD-L1 antibody achieves a synergistic inhibitory effect on PM. We highlight the importance of macropinocytosis mediated uptake of NETs in GC immune escape and PM recurrence, providing a potential combination therapeutic strategy for patients with PM.

Immune evasion driven by the tumor microenvironment is a major obstacle to effective immunotherapy in gastric cancer. To overcome this barrier, a detailed understanding of the mechanisms by which gastric cancer circumvents antitumor immunity is essential. In this study, we identified KDM5B as a pivotal epigenetic-metabolic orchestrator of immune evasion in gastric cancer. Clinically, KDM5B overexpression correlated with poor prognosis and diminished CD8+ T-cell infiltration. Mechanistically, KDM5B suppressed NLRP3-dependent pyroptosis via H3K4me3 demethylation and inhibited JAK1-STAT1/3-driven chemokine production. Crucially, a lactate-KDM5B feedforward loop facilitated immunosuppression, with KDM5B promoting lactate production and K868 lactylation enhancing KDM5B activity. Therapeutically, dual targeting of KDM5B and glycolysis overcame anti-PD-L1 resistance. Overall, these findings establish KDM5B as a central hub integrating epigenetic reprogramming, metabolic rewiring, and immunosuppression to drive progression and immune escape, positioning KDM5B as an actionable therapeutic target for reversing immunotherapy resistance in gastric cancer. KDM5B regulates an epigenetic-metabolic axis of immune evasion in gastric cancer that can be targeted to overcome anti-PD-L1 resistance, providing an effective combination immunotherapeutic strategy.

The trends and contributors of racial and ethnic disparities in causes of death among patients with clear cell renal cell carcinoma remain unclear. We analyse SEER data (2000-2019) and find that Black patients have the highest 5-year cumulative incidence of death (22.4% vs. 21.7% in whites). Despite a decreasing trend in mortality, the disparities persist between Black and white people (HR 1.15, 95% CI 1.09-1.22) and between AIAN and white people (1.25, 1.10-1.43). Disparities in death from cardiovascular disease between Black and white people increase over time. Stage at diagnosis, receipt of surgery, income, and geographic factors partially explain the observed disparities in mortality patterns. This study identifies specific, measurable contributors to mortality disparities, suggesting that interventions targeting earlier detection, equitable treatment, and socioeconomic barriers are needed.

We performed a systematic review and meta-analysis to evaluate the diagnostic accuracy, safety, and clinical impact of renal tumor biopsy (RTB) in managing renal cell carcinoma. MEDLINE, Embase, and Cochrane CENTRAL were systematically searched for studies evaluating diagnostic accuracy, complications, and clinical impact of RTB. Diagnostic performance was synthesized using hierarchical bivariate random-effects models; concordance between biopsy and surgical specimens was assessed using pooled proportions that were estimated using random-effects meta-analysis. Risk of bias was evaluated with QUADAS-2. Overall, 103 studies were included. The pooled diagnostic yield of RTB was 90%. Summary sensitivity and specificity for malignancy were 99% and 85%, respectively. Concordance between RTB and surgical pathology was high for benign versus malignant (96%) and histological subtype (89%), but moderate for grade (four-tier: 64%; two-tier: 78%). The overall complication rate was low (3%), with negligible (<1%) Clavien-Dindo ≥3 complications and 1% bleeding events (with only two patients requiring embolization). Tumor seeding was reported only in one study with a pooled seeding proportion of 0%. RTB findings was associated with a change in clinical management in 39% of patients and avoidance of active treatment in 28%. Most studies were at high risk of bias, mainly due to patient selection and flow/timing, and were predominantly retrospective. RTB is safe and high-yield, with excellent sensitivity, high concordance for benign vs malignant and histotype, but only moderate reliability for tumor grade. RTB is therefore an important aid in refining diagnosis in patients with a renal mass and supporting individualized decision-making.

Tertiary lymphoid structures (TLSs) are associated with improved responses to immune checkpoint blockade across solid tumours1,2, but how they impact the phenotypic properties of tumour-specific T cells remains unclear. Here we found, across 24 treatment-naive renal cell carcinoma (RCC) tumours, that TLS-containing tumours are more heavily infiltrated by exhausted CD8+ T cells and have a reduced terminal exhaustion transcriptional program compared with TLS- tumours. Specificity screening of 554 T cell clonotypes expanded within the microenvironment of 6 RCC tumours revealed 82 TCRs that were reactive against tumour cells and/or RCC antigens. A subset of tumour-specific T cell clonotypes (12%) was enriched within TLSs, and these expressed an increased program of stem-like progenitor exhaustion, associated with favourable anti-tumour immunity. However, in 60 independent RCC tumours, macrophages within tumour margins of TLS-containing tumours had an inferred immunosuppressive phenotype and were colocalized with exhausted putative tumour-reactive T cells in a subgroup that was further analysed, therefore supporting this mode of immune evasion as a counterbalance to T cell immune pressure. Our data reveal that TLSs are reservoirs of tumour-specific T cells with stem-like progenitor features that could be leveraged by T cell immunotherapies.

Sacituzumab govitecan (SG), a trophoblast cell surface antigen 2 (TROP2)-targeting antibody-drug conjugate, is effective against solid tumors and a promising carrier for molecular imaging, yet its role in near-infrared (NIR) fluorescence imaging of metastases is unclear. Based on the dual rationale of frequent lymph node metastasis in esophageal squamous cell carcinoma (ESCC) and prevalent TROP2 overexpression, the IRDye800cw-labeled SG probe (SG-IR800) was evaluated for detecting metastases of ESCC. Cellular uptake of SG was significantly higher in TROP2-positive KYSE150 cells than in TROP2-negative EC109 cells. Both subcutaneous KYSE150 tumors and patient-derived xenograft models with high TROP2 expression demonstrated significantly higher accumulation of SG-IR800 compared to the control probe. Peritoneal metastases yielded an NIR signal in the second window (NIR-II) comparable to primary tumors, which outperformed the NIR-I signal. Metastatic lymph nodes showed higher NIR-II contrast than inflammatory nodes. This study demonstrates that the SG-IR800 probe possesses high specificity for TROP2-positive ESCC, facilitates the detection of metastatic lesions, and holds strong potential for guiding precise intraoperative navigation.

Hypoxia is a hallmark of the tumor microenvironment, but its role in immune evasion in esophageal squamous cell carcinoma (ESCC) remains to be fully elucidated. Here, we demonstrate hypoxia inversely correlated with anti-tumor immune signatures and CD8+ T cell infiltration in clinical samples and murine models. Functionally, reducing hypoxia with the agents TH-302 or PX-478 in the AKR model enhanced intratumoral CD8+ T cell infiltration and increased their expression of Granzyme B, IFNγ, and TNFα. Mechanistically, hypoxia-induced immune suppression was dependent on protein lactylation. Inhibiting lactylation reversed the hypoxic suppression of CD8+ T cell function and abrogated the hypoxia-driven transcriptional program, which involved pathways like glycolysis, TGFβ, and Notch signaling. This lactylation-dependent regulation operated by facilitating the formation of the HIF-1 transcription complex. Specifically, mass spectrometry identified lactylation at the K172 site of HIF-1α, which was crucial for its binding to HIF-1β and subsequent target gene activation. Furthermore, in a preclinical ESCC model, pharmacological inhibition of HIF-1α with PX-478 synergized with anti-PD-1 therapy, leading to superior tumor control and enhanced CD8+ T cell cytotoxicity. Our study identifies HIF-1α K172 lactylation as a pivotal mechanism of hypoxia-mediated immune escape in ESCC, suggesting a therapeutic strategy to improve immunotherapy.

Impaired antigen processing and presentation are critical for cancer immune evasion, but the underlying mechanism is not fully known. Here, we show that downregulation of Krüppel-like factor 4 (KLF4) expression during the development of cancer is essential in this immuno-decisive process. We find that suppressed KLF4 expression causes diminished MHC class I production in esophageal cancer, forming immunosuppressive niches that limit CD8⁺ T cell infiltration. Mechanistically, KLF4 loss reduces chromatin accessibility at the MHC class I loci and the enhanceosome complex formation, suppressing MHC class I expression. In mouse allografts, KLF4 loss promotes tumor progression and immunotherapy resistance. Extending beyond esophageal cancer, we reveal that low KLF4 levels are significantly correlated with high immunotherapy failure rates across multiple human cancer types. We demonstrate that pharmacological induction of KLF4 results in enhanced antigen presentation, increases activity of CD8⁺ T cells and improves therapeutic efficacy to PD-1 blockade in mouse models. These findings extend our knowledge of cancer immunology and provide a novel insight for improving cancer immunotherapy.

In the era of personalized medicine, the development of affordable, portable biosensors for early, noninvasive diagnosis and screening of oral squamous cell carcinoma (OSCC)-particularly in remote areas-is imperative. Conventional point-of-care (POC) diagnostics are often constrained by insufficient sensitivity, which impedes timely detection. Herein, we present an unprecedented POC electrochemical biosensor that integrates UiO-66@Fe-DNA coordination polymer-functionalized flexible electrodes with a methylene blue (MB)/G-quadruplex biobarcode technology for the detection of OSCC-associated salivary miRNA-31. UiO-66@Fe-DNA coordination polymers were successfully synthesized via a facile self-assembly of Fe-DNA hybrids on UiO-66. These polymers exhibit good electrical conductivity, high stability, and a large active surface area. Meanwhile, leveraging the strong interaction between MB and G-quadruplex, the MB/G-quadruplex biobarcode technology was innovatively deployed as both a signal amplifier and an electrochemical indicator. This novel and label-free electrochemical POC device achieved an ultralow limit of detection (LOD) of 306 aM and good specificity for miRNA-31. Preliminary clinical feasibility assessments revealed good diagnostic performance (AUC = 0.98) in differentiating OSCC patients from healthy controls. These findings underscore its substantial clinical potential for early, noninvasive, and portable OSCC diagnostics.

Optimal induction chemoimmunotherapy sequential concurrent chemoradiotherapy (cCRT) in locally advanced hypopharyngeal squamous cell carcinoma (HSCC) remains undefined. This study aimed to evaluate toripalimab plus chemotherapy sequential cCRT with nimotuzumab, followed by toripalimab maintenance in unresectable stage IVB HSCC. In this single-arm, open-label, phase 2 study, untreated patients received two 21-day cycles of induction toripalimab 240 mg (d1) plus nab-paclitaxel 230 mg/m2 (d1) and cisplatin/nedaplatin 40 mg/m2 (d1-2) sequential cCRT (helical tomotherapy; nab-paclitaxel 230 mg/m2 (d1, 22, 43)) with nimotuzumab 200 mg weekly for 7 weeks. Subsequently, toripalimab maintenance was administered for eight cycles. Primary endpoint was progression-free survival (PFS). From March 16, 2022 to April 24, 2024, 31 patients completed induction chemoimmunotherapy. With a median follow-up of 31 months, median PFS and overall survival (OS) were not reached; 2-year PFS and OS rates were 67.7% and 86.4%. The 2-year larynx preservation rate was 93.3%. 30 (96.8%) patients achieved the best objective response, with 28 (90.3%) complete responses and 2 (6.5%) partial responses. The most common grade 3-4 treatment-related adverse events (TRAEs) were radiation-induced oropharyngeal mucositis and decreased appetite (each 46.9%). No grade 5 and fatal TRAEs occurred. No difficulties in speech and swallowing functions were reported in 96.8% of patients. There was no significant association between PFS and PD-L1 combined positive score, epidermal growth factor receptor expression, tumor-infiltrating T-lymphocytes, tumor mutational burden, or genetic alterations. High baseline peripheral CD3+ and CD8+CD28+ T-cell levels were associated with favorable PFS. Exploratory biomarker analyses identified that patients without disease progression after maintenance exhibited significantly higher levels of CD8+CD28⁻ T cells with a memory phenotype and lower B-cell levels. This exploratory study demonstrated preliminary efficacy and larynx preservation with manageable toxicity, possibly supporting further evaluation of induction toripalimab plus chemotherapy sequential cCRT with nimotuzumab, followed by toripalimab maintenance in stage IVB HSCC. Peripheral lymphocytes may warrant further investigation as candidate biomarkers in locally advanced disease. NCT05860335; ChiCTR2300074672.

JCOG1212 is a multi-institutional prospective single-arm trial evaluating the efficacy and safety of superselective intra-arterial cisplatin infusion with concomitant radiotherapy (RADPLAT) for locally advanced maxillary sinus squamous-cell carcinoma (MS-SCC). We previously reported the results for the T4aN0M0 cohort (n = 64), in which RADPLAT was shown to be feasible and to provide favorable survival outcomes. This report presents the results for the T4bN0M0 cohort. Patients with previously untreated T4bN0M0 MS-SCC received weekly intra-arterial cisplatin (100 mg/m2) for seven cycles in combination with radiotherapy (70 Gy in 35 fractions). The primary endpoint was 3-year overall survival (OS), with a predefined efficacy threshold of 35% and a one-sided alpha of 5%. Secondary endpoints included event-free survival (EFS), local EFS (LEFS), clinical response, and toxicity. Sixty-four patients were enrolled from 20 institutions. The 3-year OS was 68.8% [90% confidence interval (CI) 58.1% to 77.2%], exceeding the efficacy threshold. The clinical complete response rate was 54.7%. The 3-year EFS and LEFS was 51.6% (95% CI 38.8% to 63.0%) and 57.8% (95% CI 44.8% to 68.8%), respectively. With regard to acute adverse events, neutropenia ≥ grade 3 (18.0%), mucositis ≥ grade 3 (23.0%), hearing impairment ≥ grade 2 (3.3%), and stroke ≥ grade 2 (1.6%) were observed. No grade 2 ≥ creatinine increase was observed. There were no treatment-related deaths. RADPLAT achieved favorable survival outcomes with acceptable toxicity in patients with T4bN0M0 MS-SCC. These results establish RADPLAT as a feasible and effective organ-preserving treatment and demonstrate the potential of intra-arterial chemoradiotherapy for unresectable MS-SCC.

Mitochondria-ER contact sites (MERCs) are crucial signaling hubs, but their role in anti-tumor immunity is unclear. This study revealed that the mitophagy regulator PRKN ubiquitinated CD274 at MERCs in human cervical cancer cells, a key mechanism for anti-tumor immunity. CD274 expression inversely correlated with PRKN in cervical cancer. Upon mitophagy activation, CD274 was recruited from ER to MERCs by PINK1, enhancing its interaction with PRKN. PRKN then ubiquitinated CD274 at residues K89 and K105 within its extracellular domain. Functionally, a ubiquitination-deficient CD274 mutant promoted anaerobic glycolysis and MTOR signaling, accelerating cancer cell growth. Coculture with ubiquitination-deficient CD274 mutant-expressing cancer cells increased the CD8+ T-cells' exhaustion. Single-cell RNA sequencing of mouse tumors showed the expansion of the exhausted CD8+ T cells and myeloid-derived suppressor cells (MDSCs) with ubiquitination-deficient CD274 mutation. In vivo, a ubiquitination-deficient CD274 mutant accelerated tumor growth and reduced the therapy efficacy of immune checkpoint inhibitors. Conversely, clinical sample analysis showed that CD274 localization at MERCs or its ubiquitination levels were closely associated with the improved immunotherapy efficacy. Thus, mitophagy-dependent recruitment of CD274 to MERCs for PRKN-mediated ubiquitination is a novel pathway that activates the anti-tumor immunity and improves the immunotherapy efficacy, presenting a promising strategic target for cervical cancer treatment.Abbreviations: CCCP, carbonyl cyanide m-chlorophenylhydrazone; CD, cluster of differentiation; CHX, cycloheximide; FCCP, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GZMB, granzyme B; IFNG, interferon gamma; LDHA, lactate dehygrogenase A; MAP1LC3, microtubule-associated protein 1 light chain 3; MFN2, mitofusin 2; MHC, major histocompatibility complex; MTOR, mechanistic target of rapamycin kinase; OCR, oxygen consumption rate; PBMC, peripheral blood mononuclear cell; PDCD1, programmed cell death 1; PI, propidium iodide; PINK1, PTEN induced putative kinase 1; PKM, pyruvate kinase, muscle; RPS6, ribosomal protein S6; TNF, tumor necrosis factor; TME, tumor microenvironment.

The therapeutic efficacy for cervical cancer treatment is limited by insufficient drug accumulation and penetration due to physiological barriers such as mucin-rich environments after systemic administration. Thus, developing a local drug delivery system is essential to overcome these hindrances. Active transcytosis of cancer nanomedicines holds great promise for enhancing tumor extravasation, infiltration, and antitumor activity. Herein, polyzwitterionic OPDEA-PCL was developed to encapsulate paclitaxel (PTX) into micelles, serving as an intravaginal therapy for orthotopic cervical cancer. The OPDEA-PCL/PTX micelles efficiently penetrated mucus and exhibited strong resistance to mucin fouling, thereby facilitating rapid transcytosis into tumors. Furthermore, OPDEA-PCL/PTX micelles colocalized with the mitochondria of tumor cells, reversing PTX resistance. In the orthotopic cervical tumor model, the inhibition rate of OPDEA-PCL/PTX micelles was 90.0%, more than 2-fold higher than that of free PTX. In the subcutaneous cervical cancer model which is resistant to PTX. Intravenous administration of OPDEA-PCL/PTX micelles significantly overcame PTX resistance, achieving a tumor inhibition rate of 95.2%, and extending the median survival time by more than 2-fold compared to free PTX and PEG-PCL/PTX treated groups. In summary, this approach holds great promise as a potent localized nanomedicine for cervical cancer treatment with minimal side effects.

Cervical cancer (CC) is a prevalent malignancy threatening women's health. Due to its complex pathogenesis and tumor heterogeneity, it is urgent to thoroughly investigate its pathological mechanisms. Based on the TCGA database, the expression and survival analysis of PIN1 and YAP1 were assessed in CC. To verify their roles in regulating CC, stable knockdown and overexpression cell lines were constructed in CC cell lines. In vivo and in vitro experiments were conducted to verify that PIN1 regulates YAP1's SUMOylation levels and induces ACSL4's autophagic degradation. This research shows that PIN1 binds to YAP1, promoting its SUMOylation and causing autophagy-mediated degradation of ACSL4, enabling cervical cancer cells to resist ferroptosis. In vivo xenograft models confirm these findings. This study identifies PIN1 as a key redox-balance node and presents it as a potential therapeutic target for CC ferroptosis. Inhibiting PIN1 and YAP1 can inhibit tumor progression, offering a promising approach for treating aggressive CC.

Engineering the surface chemistry of nanoparticles (NPs) via the electrostatic layer-by-layer (LbL) assembly approach is a viable strategy for developing targeted drug delivery vehicles. However, how assembly solution conditions and polymer molecular weight (MW) govern LbL-NP properties remains poorly understood. We investigated the influence of layering buffer ionic strength and polymer MW on the functional performance of PLR/PLE bilayer-coated LbL-NPs designed to promote cancer cell targeting and NP retention on the extracellular membrane to deliver the cytokine interleukin-12 (IL-12). Assembly under mild ionic strength conditions increased LbL film stability but reduced IL-12 bioavailability and lipid exchange at the NP surface, ultimately diminishing therapeutic efficacy. In contrast, highly stable LbL-NPs assembled using high-MW (HMW) polymers under low ionic strength conditions exhibited enhanced in vitro NP association, increased cancer cell surface retention, and improved tumor accumulation in vivo in a metastatic ovarian cancer model, leading to improved efficacy and long-term survival. Molecular dynamics (MD) simulations revealed that HMW polymer layering produced denser PLE outer layer architecture compared to more highly interpenetrated LMW layers, providing structural insight into the superior performance of HMW LbL-NPs. These results demonstrate that control over LbL-NP delivery performance is influenced by polymer MW and assembly conditions, with HMW polymers augmenting the performance of PLR/PLE-coated NPs for IL-12 delivery.

Notch signaling is an emerging regulator of liposarcoma (LPS), but its role in mediating communication with the tumor microenvironment (TME) is unclear. Here, we investigate how Notch activation (NICD overexpression) alters the proteomes of LPS-derived extracellular vesicles (EVs). We used quantitative mass spectrometry to profile the EV proteome in multiple contexts: cultured LPS cells, LPS tumor, circulating EVs of LPS-bearing mice, and human LPS samples. We found that Notch signaling increases the secretion of EV proteins that favor tumor progression and metastasis but suppresses immune responses in murine LPS cells. Overlapping murine and human LPS data identifies 18 proteins that are increased in LPS EVs of both species, including endotrophin as a biomarker of LPS. Functional analysis supports a role of LPS EVs in regulating gene expression and behaviors of endothelial cells in TME. Together, these data demonstrate that in addition to its known function in driving tumorigenesis, Notch signaling also regulates TME through EV secretion.

Pediatric solid high-risk malignancies mostly lack established molecular biomarkers for early detection, minimal residual disease assessment, or treatment monitoring. Challenges include small patient numbers, limited sample volumes, low tumor mutational burden, and few recurrent alterations. Within the multicenter pediatric precision oncology program INFORM, we prospectively collected liquid biopsies from 130 pediatric patients and optimized cell-free DNA isolation and analysis. Whole-genome, whole-exome, and targeted panel sequencing were performed using liquid biopsy-adapted protocols. Integrating tissue-derived molecular profiles and orthogonal validation revealed that low-coverage whole-genome sequencing reliably detects circulating tumor DNA. An in silico ctDNA estimation score, combining fragment length and genome segment alterations, improved sensitivity and specificity to 95%, enabling plasma-based tumor detection in 93% of patients. Whole-exome and panel sequencing effectively identified clinically relevant, potentially druggable molecular targets. However, their utility varied substantially across different tumor entities, underscoring the need for entity-specific considerations in the interpretation and application of these methodologies. In-depth analyses demonstrated liquid biopsy's potential to track tumor evolution, identifying common tumor ancestors and refining patient stratification. This study advances liquid biopsy methodologies in pediatric oncology and provides a rationale that, as SNVs are more sensitively captured by panel sequencing and WES, while CNVs are better represented by lcWGS and WES. The underlying tumor genomic profile should guide the selection of liquid biopsy assays to optimize clinical decision-making. Systematic liquid biopsy analyses within the pediatric precision oncology INFORM registry enabled a real-world, multicenter comparison of sequencing approaches across high-risk malignancies. By optimizing preanalytical and bioinformatic tools for pediatric settings, we improved plasma-based cancer detection, molecular tumor characterization, and identification of targetable alterations, laying the groundwork for integration into personalized medicine programs and clinical trials.

The clinical significance of one or two measurable residual disease (MRD) assessments is established in multiple myeloma (MM). However, how to stratify patients according to ≥3 MRD assessments remains unknown. The traits of MRD resistance and if treatment of persistent MRD vs relapse could improve outcomes also remains unknown. MRD dynamics were computed using next-generation flow cytometry and Connector in 539 newly-diagnosed MM patients with ≥3 assessments in the GEM2012MENOS65/GEM2014MAIN and GEM2017FIT trials. Molecular and immune profiling were performed in matched diagnostic and MRD samples. The survival impact of treating persistent MRD vs relapse was investigated with anti-BCMA CAR T cells in MIcγ1huCRBN mice. Computed MRD dynamics based on 3,610 MRD assessments identified five subgroups with different survival. Patients with late-sustained MRD response had excellent outcomes, similar to those with early-sustained MRD response. Patients with volatile results and those with primarily and resurgent MRD resistance had dismal survival. MRD dynamics outperformed transplant-eligibility and the R-ISS. These results were validated in 249 MM patients treated in routine practice. Multiomics characterization of MRD dynamics in patients and mouse models of MRD resistance revealed genomic evolution, transcriptional adaption and a pro-inflammatory tumor-immune microenvironment. Increasing clonality and exhaustion of endogenous T cells throughout disease progression urged investigating if MRD interception with anti-BCMA CAR-T cells could improve outcomes. Infusion at MRD resistance prolonged mouse survival compared to identical treatment at relapse. Altogether, MRD dynamics is the strongest predictor of progression and may help tailoring treatment to prevent additional tumor and immune alterations prior to relapse.

Gallbladder carcinoma (GBC) is a highly aggressive malignancy characterized by an extremely poor prognosis and limited therapeutic options. N6-methyladenosine (m6A), the most prevalent post-transcriptional modification in eukaryotic mRNA, plays a pivotal role in regulating tumor progression. Screening of an siRNA library targeting YTH-domain family reader proteins identified YTH domain-containing family protein 2 (YTHDF2) as essential for the malignant phenotype of GBC cells. YTHDF2 was aberrantly upregulated in GBC tissues and served as an independent predictor of poor prognosis. Genetic silencing of YTHDF2 markedly suppressed both proliferation and metastatic potential of GBC cells in vitro and in vivo. Mechanistically, YTHDF2 transcription is upregulated through P300 histone acetyltransferase-mediated H3K27ac modification. YTHDF2 promotes mRNA degradation by binding to m6A-modified sites within the transcript encoding the metastasis suppressor non-metastatic protein 1 (NME1), thereby repressing NME1 expression and accelerating GBC malignancy. Co-treatment with the P300 inhibitor C646 and the m6A modification inhibitor STM2457 synergistically inhibited YTHDF2-mediated GBC malignancy. Collectively, these findings establish the P300-YTHDF2-m6A-NME1 axis as a critical driver of GBC progression and indicate that combined C646 and STM2457 treatment represents a novel and promising therapeutic strategy for GBC.

Curative-intent surgery in patients with perihilar cholangiocarcinoma (pCCA) is associated with substantial surgical risks and high early-recurrence rates. This study aimed to develop an ABC system for preoperative staging of patients with pCCA. This retrospective international multicenter cohort study included patients with resected pCCA across 27 participating centers from 9 countries (2006-2022). The prognostic performance of the ABC system for overall survival (OS) and recurrence was assessed using multivariable (cause-specific) Cox regression. Among 1307 included patients (median age, 66 [IQR, 57 to 73] years), independent prognostic factors for OS were: tumor size ≥25 mm (adjusted hazard ratio [aHR], 1.32 [95% CI, 1.13 to 1.55]; P=0.0003), CA19-9 ≥500 U/mL (aHR, 1.49 [1.22 to 1.81]; P<0.0001), and WHO performance status ≥1 (aHR, 1.35 [1.12-1.63]; P=0.002). The ABC score for OS consisted of 1 point for each independent prognostic factor (0-3 points); the ABC score for recurrence did not include WHO PS ≥1 (0-2 points). Compared to the ABC-0 group, the highest ABC risk group had a 3.4 times higher 90-day mortality risk (21 vs 6%; P=0.005), a 3.3 times higher 6-month recurrence rate (18 vs 5%; P<0.0001), a 3.0 times shorter median OS (13 vs 39 months; P=0.0001), and a 3.6 times lower 5-year OS rate (11 vs 35%; P=0.018). The prognostic value of the ABC score for OS remained similar in a sensitivity analysis including only patients with a resection post-2015 (Pinteraction=0.90). Following the ABC system, we should be reluctant to offer resection to patients with an ABC score of 3 with a tumor size ≥25 mm, CA19-9 ≥500 U/mL, and a WHO PS ≥1.

TIGIT (T cell immunoreceptor with Ig and ITIM domains) has emerged as a key exhaustion marker of intratumoral natural killer (NK) cells, but results from clinical trials with TIGIT blockade have been largely negative. Recent data have suggested that a subset of TIGIT-expressing NK cells can show increased functionality. We hypothesized that there are differences in function between peripheral and intratumoral TIGIT-expressing NK cells in patients with sarcoma and preclinical sarcoma models, which undermine the efficacy of systemic TIGIT blockade. We sought to investigate differences in TIGIT+ NK cells using systemic versus intratumoral TIGIT-blocking strategies. Peripheral and intratumoral NK cells were analyzed from human patients and mice with osteosarcoma (OSA) and soft tissue sarcoma (STS). NK phenotype and function were evaluated using flow cytometry, immunohistochemistry, live-cell imaging, and RNA sequencing. Mouse antimouse-IgG1 TIGIT blockade was delivered systemically or intratumorally in flank models of OSA (K7M2) and STS (MCA-205). Clinical and genomic data were evaluated using Caris CODEai. Expression of the TIGIT ligand CD155 on myeloid and tumor cells was evaluated as a marker of TIGIT function. Using multiple readouts, TIGIT+ NK cells from the spleen of tumor-bearing mice or peripheral blood of patients with STS and OSA showed increased functionality compared with TIGIT- NK, while TIGIT+ NK cells from the sarcoma tumor microenvironment (TME) of mice and humans were dysfunctional, with upregulated senescence and inhibitory gene pathways. Systemic TIGIT blockade reinvigorated intratumoral NK cell function but inhibited peripheral NK cells, while intratumoral administration of TIGIT blockade significantly delayed tumor growth and prolonged survival with partial reversal of NK dysfunction in the TME. Clinical and genomic data demonstrated that more NK cell infiltration in human STS was prognostic of improved overall survival only when intratumoral CD155 expression was low, indicating that elevated CD155 expression is correlated with greater NK cell dysfunction. We identified a cross-species role of TIGIT expression dependent on location, where peripheral TIGIT+ NK cells showed evidence of enhanced functionality, while intratumoral TIGIT+ NK cells were dysfunctional. These differences impacted antitumor effects of TIGIT blockade, suggesting that intratumoral delivery of TIGIT blockade may be a novel translational strategy in high-risk bone and soft tissue sarcomas.

Inborn errors of immunity (IEI) are monogenic disorders of the immune system that lead to immunodeficiency, autoimmunity, autoinflammation, allergy, and/or cancer. This review provides a comparative review of the epidemiology, clinical manifestations, and management of IEI in China. While individual forms of IEI are generally rare disorders globally, collectively they represent a significant disease burden. China has recently made great strides in the diagnosis and treatment of IEIs with the increasing utility of next-generation sequencing and the availability of hematopoietic stem cell transplantation. Challenges remain in the establishment of national registry databases comparable to the European Society for Immunodeficiencies and The United States Immunodeficiency Network. Also, the absence of a national newborn screening program for severe combined immunodeficiency poses risks regarding live attenuated vaccination in undiagnosed infants. Current access to treatment including subcutaneous immunoglobulin, targeted molecular therapies, and commercialized gene therapy remains limited in China. Nevertheless, the therapeutic landscape is rapidly evolving, marked by an increasing clinical application of precision molecular-targeted treatments and the emergence of AI-empowered gene editing therapies. Ultimately, advancing the field of IEI necessitates global collaboration, integrating the complementary strengths of both regions to establish scientific breakthroughs and clinical innovation.

Dapagliflozin improves kidney outcomes in patients with chronic kidney disease (CKD), yet the underlying mechanisms by which it exerts protective effects are not fully elucidated. Here, we applied plasma proteomics to identify proteins and pathways linked to CKD progression and examined which of these are modulated by dapagliflozin. We measured 3072 circulating plasma proteins (Olink, Upsala, Sweden) in 2485 (57.7%) of the 4304 patients with CKD from the DAPA-CKD trial (eGFR 25-75 mL/min/1.73m2 ; and urinary albumin to creatinine ratio 200-5000 mg/g). Proteins associated with the composite kidney outcome (50% eGFR decline, kidney failure, and kidney death) and with dapagliflozin treatment were respectively identified by multivariable Cox proportional hazard regression and ANCOVA. Ingenuity pathway analysis was used to identify relevant molecular pathways and upstream regulators. Pathways that had the strongest association with the composite kidney outcome were hepatic fibrosis, insulin-like growth factor transport regulation, and tumor necrosis factors signaling. These pathways were generally related to fibrosis and inflammation. After 12 months dapagliflozin treatment showed a (placebo corrected) modification of 216 proteins, of which kidney injury molecule-1 showed the strongest reduction (-14.9%; 95% confidence interval-17.9, -11.7). Of the 137 pathways associated with the kidney outcome, 35 (25%) were modified by dapagliflozin. Among these, activity of 11 (31% of 35) pathways decreased, and 10 (29% of 35) pathways were increased. Pathways associated with kidney outcome and modulated by dapagliflozin were related to extracellular matrix remodeling, inflammation and fibrosis, and immune-vascular interactions. This proteomic analysis reveals mechanisms underlying dapagliflozin's beneficial effects in CKD progression and shows that key pathways associated with CKD progression are modified by dapagliflozin, supporting its anti-inflammatory and anti-fibrotic potential.

Fistulising perianal Crohn's disease (CD) remains a debilitating condition, with few randomised controlled trials conducted so far. To evaluate the efficacy and safety of ustekinumab (UST) in patients with CD with active draining perianal fistulas (NCT04496063) in a randomised placebo-controlled trial. In this double-blind, multicentre Groupe d'Etudes Therapeutiques des Affections Inflammatoires Digestives trial, patients enrolled with active fistulising perianal CD were randomised 1:1 to receive UST (6 mg/kg intravenously at baseline followed by 90 mg subcutaneously at week 8, then every 8 weeks) or placebo, after standardised surgical management. The primary endpoint was combined clinical remission (absence of drainage from all external fistula openings) and radiological remission (absence of abscesses >2 cm on blinded central MRI) at week 12. At week 12, placebo non-responders could switch to UST, and UST non-responders could be intensified during the open-label phase. 32 patients were randomised (UST: 16; placebo: 16) across 10 French centres and 69% had prior anti-tumour necrosis factor failure. At week 12, combined remission was achieved in 62% with UST versus 25% with placebo (OR=5.1 (95% CI 1.07-24.4)). Clinical remission occurred in 62.5% vs 31% (OR=3.75 (95% CI 0.84 to 16.8)) and radiological remission in 87.5% vs 75% (OR=2.7 (95% CI 0.34 to 21.1)). Nine placebo-patients switched to UST during the open-label phase. Combined remission rates at week 48 were 50% in the placebo arm and 31% in the UST arm, respectively. These findings suggest greater short-term clinical benefit with UST compared with placebo and support its use in patients with active fistulising perianal CD. NCT04496063.

To evaluate the safety and effectiveness of eye-preserving therapies in patients with American Joint Committee on Cancer (AJCC) eighth edition cT3c retinoblastoma presenting with neovascular glaucoma (NVG) without buphthalmos (defined as early cT3c), focusing on overall survival and eye preservation. Retrospective, single-center cohort study. 132 patients diagnosed with early cT3c retinoblastoma from May 2014 through October 2024. The patients were divided into primary enucleation (50 patients) and primary eye-preserving groups (82 patients). They were followed up for survival status and ocular outcomes. Overall survival, high-risk pathological features, globe salvage and vision preservation. After a median follow-up of 52.9 months, one death occurred in each group, and overall survival did not differ significantly between the primary eye-preserving and primary enucleation groups (log-rank test, P = 0.775). Eye-preserving therapies were associated with a lower incidence of high-risk pathological features (odds ratio [OR], 0.21; P = 0.003), with attenuated severity of both choroidal (OR, 0.25; P = 0.002) and optic nerve invasion (OR, 0.23; P = 0.008). The globe salvage rate was 49.4% (41/83) in the primary eye-preserving group. And among these preserved eyes, 46.3% (19/41) regained light projection or better after receiving eye-preserving therapies. Importantly, presenting intraocular pressure (IOP) ≥32 mmHg (hazard ratio [HR], 2.37; P = 0.010) and corneal edema (HR, 2.86; P = 0.007) were high risk factors for globe salvage failure. Compared with intravenous chemotherapy (IVC) alone, application of intra-arterial chemotherapy (IAC; HR, 0.13; P = 0.001) alone and combined IVC-IAC regimens (HR, 0.15; P = 0.001) demonstrated a significantly association with better globe salvage outcomes. Additionally, cryotherapy (HR, 0.14; P < 0.001) was identified as an independent protective factor for overall globe salvage. Primary eye-preserving therapies can secure high rates of globe salvage with partial visual function in patients with early cT3c retinoblastoma, without jeopardizing patient survival. However, eyes exhibiting corneal edema or IOP ≥32 mmHg demonstrate markedly inferior salvage outcomes, the treatment strategy for such patients must therefore be cautiously individualized.

Previous studies of risk factors for subsequent soft-tissue sarcoma (STS) among childhood cancer survivors had small numbers and were unable to comprehensively investigate the dose-response relationships with radiation from radiotherapy and with cumulative exposure to specific cytotoxics. We conducted a nested case-control study, encompassing 275 subsequent STS cases and 275 matched controls, within the Pan-European cohort of 69 460 5-year survivors from 12 countries. Odds ratios (ORs) and 95% confidence intervals (CIs) for subsequent STS were calculated for different levels of radiation dose to the STS location (in Gy) and for cumulative doses of specific chemotherapeutic agents (in g/m2). Additionally, excess ORs per Gy (EOR/Gy) or per g/m2 (EOR/g/m2) were calculated to assess dose-response relationships. The OR for subsequent STS was 22-fold [95% CI 6.9-95.4] higher in soft tissue exposed to ≥30 Gy and remained in excess with exposure to 5-9 Gy [OR = 3.8, 95% CI 1.3-12.0] compared with no radiation. The EOR/Gy was 0.86 [95% CI 0.35-2.16], with a particularly high risk observed in survivors of neuroblastoma [EOR/Gy = 5.52, 95% CI 0.71-52.31] or bone sarcoma [EOR/Gy = 4.16, 95% CI 0.40-24.72], and in females [EOR/Gy = 2.35; 95% CI 0.65-8.14]. For patients who had received a cumulative procarbazine dose of ≥6.0 g/m2, the OR was 4.7 [95% CI 1.3-25.1] compared with non-exposure after controlling for radiation. No association was found for other alkylating agents or other specific cytotoxic drugs. Although high radiation doses remain the primary risk factor for secondary STS, our findings suggest a possible increase in risk at lower doses (5-9 Gy) and following procarbazine treatment among childhood cancer survivors. These observations warrant further investigation and may merit consideration in treatment planning and long-term follow-up guidelines for cancer survivors.

Cardiovascular-kidney-metabolic (CKM) syndrome, formally defined by the American Heart Association in 2023, affects approximately 90% of US adults, who meet criteria for stage 1 or higher. The rapid convergence of multiple drug classes on CKM pathways-SGLT2 inhibitors, finerenone, GLP-1 receptor agonists, ARNI, and interleukin-directed therapies-has created an urgent need for pharmacologically grounded frameworks that guide drug selection, interpret biomarker responses, and monitor target engagement across interconnected organ systems. This review proposes a three-dimensional biomarker-guided approach to precision pharmacotherapy in CKM syndrome. In the organ-specific dimension, we map key biomarkers to their corresponding drug targets and elucidate the molecular mechanisms underlying drug-biomarker interactions: SGLT2 inhibitors attenuate myocardial injury through metabolic substrate shifting toward ketone body utilization and, based on preclinical evidence, NHE1 inhibition; neprilysin selectivity of sacubitril/valsartan explains the differential natriuretic peptide response; and tubuloglomerular feedback mediates the renoprotective hemodynamic effects of SGLT2 inhibitors. In the pathway-specific dimension, we identify cross-system biomarkers-hs-CRP, IL-6, galectin-3, GDF-15, and FGF21-that reveal shared druggable targets spanning the IL-1β/NLRP3 inflammasome axis (canakinumab, colchicine), IL-6 trans-signaling (ziltivekimab), and FGF21/β-klotho metabolic signaling. In the temporal dimension, we demonstrate how serial biomarker trajectories serve as pharmacodynamic readouts that distinguish therapeutic drug effects from disease progression, including the initial eGFR dip with SGLT2 inhibitors and natriuretic peptide changes during combination therapy. Central to this framework is the concept of "pharmacological phenotyping"-using multi-biomarker panels to define drug-responsive pathophysiological states that directly inform therapeutic selection, analogous to companion diagnostics in oncology. We further present a comprehensive drug-biomarker interaction matrix with pharmacological rationale and analyze the emerging drug development pipeline, including RNA-based Lp(a) therapeutics, FGF21 analogues, galectin-3 inhibitors, and in vivo CAR-T anti-fibrotic approaches. This framework provides a practical roadmap for biomarker-guided precision pharmacotherapy in CKM syndrome.

Sarcopenia is a progressive muscle-wasting condition driven in part by chronic inflammation; however, the specific inflammatory mediators that contribute to functional decline in humans and their responses to exercise remain poorly defined. We analyzed public ribonucleic acid (RNA) datasets to identify inflammatory pathways associated with aging and exercise. Additionally, we validated candidate factors in older adults through 2 human studies. Complementary cellular and animal experiments were performed to dissect the mechanistic responses to muscle atrophy, injury, and exercise-mimetic therapies. C-X-C motif chemokine ligand 2 (CXCL2) was identified as a prominent age-associated exercise-responsive inflammatory factor in human datasets. In older adults, plasma CXCL2 levels increased with sarcopenia severity and correlated with SPS-defined functional impairment (r = 0.6472, p = 0.0003). In the 48-week human intervention, structured exercise substantially improved strength, mobility, and endurance and markedly reduced circulating CXCL2 (p < 0.0001) and tumor necrosis factor-alpha (TNF-α) levels (p < 0.0001). In contrast, individuals in the non-exercise group exhibited progressive functional decline and increased cytokine levels. Moreover, Cxcl2 expression was upregulated in the skeletal muscles of aged mice and induced by muscle atrophy or injury (p = 0.0082). In contrast, exercise-mimetic stimulation and acute endurance exercise suppressed Cxcl2 expression (p = 0.0148, p = 0.0072). Importantly, blocking CXCL2-CXCR2 signaling attenuated dexamethasone-induced expression of the atrophy markers Fbxo32 (p = 0.0336, 0.0127, 0.0061) and Trim63 (p = 0.0296, 00160, 0.0042). Human evidence from year-long clinical exercise interventions, supported by transcriptomic, cellular, and animal experiments, identified CXCL2 as a modifiable inflammatory mediator linking aging to muscle atrophy. Consistent suppression of CXCL2 by exercise and attenuation of atrophic signaling through CXCR2 inhibition highlights the CXCL2-CXCR2 axis as a promising target for mitigating sarcopenia.

Advanced heart failure (AdvHF) remains a major clinical challenge, yet contemporary epidemiological real-world data on AdvHF patients are limited. Aim was to assess 20-year trends in clinical characteristics, outcomes, and use of left ventricular assist devices (LVAD) and heart transplantation (HTx) in patients with AdvHF and reduced ejection fraction (AdvHFrEF) in Sweden. Between 2003-2022, 5,323 patients (median age 76 [IQR 68-82], 23% females) included in the Swedish HF Registry met adapted ESC-HFA criteria for AdvHF (NYHA III-IV, EF<30%, ≥1 HF hospitalizations ≤6 months). Of these, 29% were eligible for AdvHF therapies (median age 63 [IQR 57-67] years, 18% females) according to the following additional criteria: ≤70 years old, no dialysis within ≤5 years, no active cancer within ≤3 years.Unadjusted temporal trends in 1-year outcomes (all-cause, CV and non-CV death, HTx and LVAD implantation) were presented as estimated annual percent change (eAPC). 1-year all-cause mortality declined significantly in the overall cohort (eAPC -3.8; 95%CI -5.3 to -2.2) as well as in patients eligible for AdvHF therapies (eAPC -3.2; 95%CI -6.3 to -0.1) over the study period. These improvements were largely driven by reductions in CV mortality (overall cohort: eAPC -4.3; 95%CI -6.0 to -2.6; eligible cohort: eAPC -5.6, 95%CI -9.4 to -1.7). Use of LVAD significantly increased over the study period, while HTx showed a modest but non-significant rise. In a large nationwide cohort, all-cause and CV-mortality among AdvHFrEF patients markedly declined over the past 20 years, potentially reflecting improvements in treatments. Despite progress, significant opportunities remain to improve access to and adoption of advanced therapies.

Ritlecitinib, an oral Janus kinase (JAK) 3/tyrosine kinase expressed in hepatocellular carcinoma (TEC) family kinase inhibitor, demonstrated safety in patients aged 12 years and older with alopecia areata (AA) in an initial integrated analysis of 4 clinical studies for up to 2 years. This updated integrated safety analysis evaluated the safety of ritlecitinib up to ~ 5 years in patients aged ≥ 12 years with AA from the ALLEGRO clinical trial program. Safety data were pooled from 4 studies. Two groups were analyzed: patients who received any dose of ritlecitinib (30 mg or 50 mg with or without a 4-week 200 mg loading dose, or 10 mg daily) ("any-ritlecitinib" group) and a subset of the any-ritlecitinib group including only patients who received ritlecitinib 50 mg daily with or without a 4-week 200 mg daily loading dose (ritlecitinib 50 mg ± 200-mg group). Safety data were summarized descriptively. Proportions and incidence rates (IRs; IR/100 patient-years [PYs]) of adverse events (AEs) were evaluated. In the ritlecitinib 50-mg ± 200-mg (N = 1228) and any-ritlecitinib (N = 1294) groups, median duration of exposure was 1197 days (3261.5 PYs) and 1204 days (3539.5 PYs), respectively. AEs occurred in 1070 patients (87.1%; 148.1/100 PYs) in the 50-mg ± 200-mg group and 1158 patients (89.5%; 167.9/100 PYs) in the any-ritlecitinib group. The most common AEs included headache, positive SARS-CoV-2 test, and nasopharyngitis. Serious AEs were reported in 6.8% of patients (2.6/100 PYs) in the 50-mg ± 200-mg group and 6.8% of patients (2.5/100 PYs) in the any-ritlecitinib group. There were two deaths. Overall, 8.1% of patients (3.0/100 PYs) and 8.4% of patients (3.0/100 PYs) in the 50-mg ± 200-mg and any-ritlecitinib groups, respectively, had an AE that led to discontinuation from the study or study drug. IRs for both groups were 0.1/100 PYs for opportunistic infections, 1.0/100 PYs for herpes zoster, 0.3/100 PYs for malignancies (excluding nonmelanoma skin cancer), and 0.2/100 PYs for major adverse cardiovascular events. In this updated integrated safety analysis of the ALLEGRO clinical trials, long-term ritlecitinib treatment was generally well tolerated up to ~ 5 years in patients aged ≥ 12 years with AA. The overall safety profile was consistent with previously reported data. NCT03732807, NCT04006457, NCT04517864, NCT02974868. Video Abstract Updated integrated safety analysis of ritlecitinib up to ~ 5 years in patients with alopecia areata from the ALLEGRO clinical trial program (MP4 407314 KB).

Lymphatic endothelial cells (LECs) line the lymphatic vasculature and support interstitial fluid drainage, lipid transport, and immune-cell trafficking. Beyond these classical functions, recent studies now recognize LECs as heterogeneous, spatially organized endothelial regulators that maintain lymphatic identity while adopting tissue-, segment-, and disease-associated states. Advances in lineage tracing, multiomics, functional imaging, and perturbation studies now link LEC heterogeneity to drainage control, immune surveillance, antigen handling, metabolic homeostasis, and tissue repair. Despite these advances, a unified framework explaining how developmental origin, tissue niche, and context-dependent state transitions collectively shape LEC function across physiology and disease remains lacking. In this review, we summarize the developmental origins, identity-maintenance mechanisms, and anatomical deployment of LEC states across the lymphatic network. We then discuss how specialized LEC states coordinate lymphatic transport, immune surveillance, and peripheral tolerance under homeostasis. We further examine how context-dependent LEC reprogramming contributes to lymphatic disorders, inflammation and autoimmunity, cancer, cardiometabolic disease, and central nervous system dysfunction. Finally, we highlight challenges and opportunities for translational LEC-targeted therapy. Together, this state-centered framework reframes LECs as actionable regulators of tissue homeostasis and disease progression, providing a conceptual foundation for precision lymphatic medicine.

Coronary microvascular dysfunction (CMD) is a key contributor to myocardial ischemia and cardiovascular diseases. It is characterized by abnormalities of the coronary microvasculature, leading to impaired myocardial perfusion. Although CMD has a high prevalence in patients with nonobstructive coronary artery disease and is associated with adverse cardiovascular events, its pathogenesis has not yet been fully elucidated. Current diagnostic approaches combine noninvasive and invasive methods, but there remains a lack of effective targeted therapies. This review discusses the epidemiology, pathophysiology, diagnostic strategies, and treatment options for CMD, with a particular focus on the protective role of the neuregulin-1 (NRG-1)/v-erb-b2 erythroblastic leukemia viral oncogene homolog B (ErbB) signaling pathway. We initially outline how the NRG-1/ErbB pathway affects endothelial function, ventricular remodeling, oxidative stress, and myocardial angiogenesis, highlighting its potential as a therapeutic target. In addition, we explore emerging evidence that traditional Chinese medicine (TCM) interventions may regulate the NRG-1/ErbB axis to improve microvascular function and cardiac outcomes. Overall, this review deepens our understanding of the mechanisms underlying CMD and provides new avenues for integrated precision therapies, including TCM, with the aim of improving clinical management and prognosis in patients with CMD.

Rhabdomyosarcoma (RMS) is the most common soft tissue malignancy in children and adolescents and is characterized by expression of myogenic regulatory factors without terminal differentiation. Treatment of RMS remains challenging, with nearly one-third of patients experiencing relapse due to chemotherapy resistance. However, the molecular mechanisms underlying RMS growth and chemoresistance remain poorly understood. Fibroblast growth factor-inducible 14 (FN14), a receptor for the TWEAK cytokine, has been implicated in tumor progression in several cancers, but its role in RMS is unknown. Here, we demonstrate that FN14 expression is highly upregulated in human RMS samples and in the RD and RH30 RMS cell lines. Silencing FN14 reduced proliferation and survival of RMS cells. In a xenograft model, inducible FN14 knockdown transiently delayed tumor growth, supporting a functional role for FN14 in RMS growth in vivo. FN14 expression was further increased in vincristine-resistant RD cells, and FN14 knockdown suppressed the proliferation and survival of these chemoresistant cells. Mechanistic studies performed primarily in RD cells showed that FN14 promotes ERK1/2, but not p38 MAPK, signaling. Consistent with this finding, pharmacological inhibition of ERK1/2 reduced RD cell proliferation and survival. Furthermore, both FN14 silencing and ERK1/2 inhibition decreased mitochondrial oxidative phosphorylation in RD cells. Finally, inhibition of either FN14 or ERK1/2 promoted myogenic differentiation in parental and vincristine-resistant RD cells. Collectively, our experiments support FN14 as an important regulator of proliferation, survival, chemoresistance, and differentiation in RMS and support further investigation of FN14-mediated signaling in RMS pathogenesis.

Immunotherapy has revolutionized the treatment of solid cancers in recent years. However, T-cell lymphomas (T-NHLs) originate from immune cells themselves and are biologically heterogeneous, rendering investigations of immune checkpoint inhibitor (ICI) mechanisms of action complex. While case reports and individual Anaplastic Large Cell Lymphoma (ALCL) cases enrolled in T-NHL trials demonstrated favourable responses to ICI, hyperprogression was observed in other T-NHL subtypes. We therefore utilized a syngeneic mouse model of ALK+ ALCL to investigate immune surveillance and ICI-induced immune response. Transplantation experiments combined with depletion of relevant immune axes revealed that ALCL immune surveillance is mediated by CD4+ T cells and NK cells. Innate and adaptive immune cell infiltration was confirmed on a large series of primary human ALK+ ALCL samples. ICI monotherapy demonstrated robust efficacy in murine ALCL, inducing complete remissions in approximately 50% of treated animals. Mechanistically, PD-L1 blockade reversed Treg-mediated immunosuppression and increased the frequency of circulating effector CD8+ T lymphocytes, thereby prolonging survival significantly. Importantly, CD4+ T cells proved indispensable for driving and sustaining immunotherapy-induced anti-tumour responses in murine ALCL. CD4+ T cells of non-responder animals exhibited an exhausted phenotype and a transcriptomic Th22-like signature, implicating persistent T-cell exhaustion and polarization as a meaningful immune-evasion mechanism. Our findings uncover CD4+ T cells as key players in spontaneous and immunotherapy-mediated anti T-cell lymphoma immunity, which demonstrates the critically needed preclinical proof-of-concept for the safe and effective use of immunotherapy for ALCL.

The intricate balance between angiotensin-converting enzyme 1 (ACE1) and 2 (ACE2) in the pulmonary vasculature is pivotal for the pathogenesis of pulmonary arterial hypertension (PAH). Catalysing the K48-linked deubiquitination, ubiquitin carboxyl-terminal hydrolase 10 (USP10) is involved in tumour suppression, autophagy, and cell proliferation. This study aims to determine whether a positive feedback loop of USP10 and AMP-activated protein kinase (AMPK) in pulmonary endothelium is protective against PAH. In silico data analyses and in vitro culture cell experiments were used to investigate the role of USP10 in human idiopathic PAH (IPAH) and rodent pulmonary hypertension (PH) as well as the underlying mechanism involving a positive feedback loop of AMPK and USP10 in lung endothelium. Endothelial cell (EC)-specific USP10 transgenic (Tg) mice and mice administered liraglutide were used to explore the efficacy of the AMPK/USP10 loop in mitigating PH in rodents. USP10 level was decreased in the lung endothelium of human IPAH and rodent PH. AMPK/USP10 loop activation increased ACE2 Ser-680 phosphorylation and Lys-788 deubiquitination, thus contributing to the homeostatic level of ACE2 and lung vascular patency. Mice with liraglutide administration phenocopied the mitigated PH in EC-specific USP10 Tg mice, in part because of the activated AMPK/USP10 loop in the pulmonary endothelium. Genetic or pharmacologic [via glucagon-like peptide-1 receptor agonists (GLP-1 RAs)] interventions in the AMPK/USP10 loop can augment ACE2 level in lung endothelium. This type of ACE2 enhancement garners protection against PAH in humans and PH in rodents, which provides a rationale for using GLP-1 RAs to alleviate PAH.

Bone metastases are refractory to current therapies, primarily owing to the immunosuppressive metastasis tumor microenvironment (TME), which is dominated by myeloid cells. However, the function and regulatory mechanisms of myeloid compartments within the bone TME are incompletely understood. Herein we sought to delineate the role of the DKK1-CKAP4 axis in shaping myeloid cell-mediated immunosuppression in bone metastases and to identify potential therapeutic strategies targeting this pathway. The composition and phenotypic characteristics of myeloid cells in the bone TME were analyzed. Mechanistic studies were conducted using ex vivo co-culture systems and an in vivo Ckap4fl/flS100a8Cre (neutrophil-specific Ckap4 knockout) mouse model to delineate the role of the DKK1-CKAP4 axis in regulating neutrophil maturation, osteoclast-like cell differentiation, and macrophage polarization. The therapeutic efficacy of DKK1 blockade in combination with zoledronic acid, as well as the impact on remodeling the TME, was further validated in bone metastasis mouse models. Significant expansion and predominance of myeloid cells within the bone TME was noted. Mechanistically, the DKK1-CKAP4 axis regulated the development and function of multiple myeloid populations. The DKK1-CKAP4 axis drove neutrophils toward an immature-like, immunosuppressive phenotype, promoted osteoclast-like cell differentiation, and induced macrophage polarization into an M2-like phenotype. Importantly, combined therapy with DKK1 blockade and zoledronic acid reprogrammed immunosuppressive myeloid cells, restored antitumor immunity, and significantly reduced tumor burden in bone metastasis models. The findings herein showed the DKK1-CKAP4 axis to be a key regulator of myeloid-driven immunosuppression in the bone TME. The combination of DKK1 blockade with zoledronic acid represents a potential therapeutic strategy for bone metastases.

Neuroblastoma (NB) is the most prevalent pediatric solid tumor, associated with a poor prognosis. While alternative polyadenylation (APA) is a key post-transcriptional regulatory mechanism in many cancers, its role in NB remains largely unexplored. We used public expression profiles from NB and normal samples, along with single-cell transcriptomic data from our MYCN-transgenic mouse models, to screen NB-upregulated APA related genes. Knockdown method was used to examine the function of NUDT21 in NB cells and in nude mice. RNA-seq and PAS-seq were performed to identify differentially expressed genes with APA events. CUT&Tag was used to analyze transcriptional activation of downstream genes. NUDT21 was found substantially upregulated in NB, driven by NB master transcription factors (TFs) like MYCN. NUDT21 knockdown inhibited NB cell proliferation, induced cycle arrest and apoptosis in vitro, and suppressed xenograft growth in nude mice. Mechanistically, NUDT21 silencing triggered global APA remodeling, with approximately 85% of APA genes shifting to proximal poly(A) sites. Integrative PAS-seq and RNA-seq analyses revealed that NUDT21 regulated E2F target genes (e.g., MKI67) via APA, promoting longer 3' UTR isoforms to enhance mRNA stability and translation. CUT&Tag analysis further indicated direct transcriptional regulation on E2F target genes by NUDT21 through promoter binding. This convergence of APA-mediated post-transcriptional regulation and transcriptional activation on the E2F network established a sophisticated oncogenic paradigm. Our study identified NUDT21 as a critical driver of NB progression through both transcriptional and post-transcriptional mechanisms, laying the foundation for NUDT21-targeted NB therapies.

Triterpenoids are important natural secondary metabolites with diverse bioactivities, including antioxidant, anti-inflammatory, and anti-cancer properties, making them valuable for applications in the pharmaceutical, cosmetic, and food industries. Currently, triterpenoids are mainly obtained through natural extraction or chemical synthesis. However, these conventional approaches are often limited by production efficiency, environmental burdens, and product diversity. Rapid advances in metabolic engineering and synthetic biology have promoted the emergence of heterologous biosynthesis as a promising, efficient, and sustainable strategy for triterpenoids production. In this review, we first summarize the classification and bioactive properties of triterpenoids, together with the challenges and potential solutions associated with their microbial synthesis. Then, we analyze the key characteristics of microbial hosts and their corresponding biosynthetic pathways for triterpenoids production, aiming to establish programmable platforms that overcome the limitations of natural biosynthesis. Subsequently, we propose metabolic engineering and synthetic biology strategies, including enzyme optimization, pathway optimization, compartmentalization engineering, and systems biology approaches, for optimizing matter and energy transmission and thereby enhancing triterpenoids production. We further discuss the potential challenges for scaling up triterpenoids production from laboratory-scale studies to industrial-scale applications, including the optimization of large-scale fermentation process and the improvement of downstream extraction and recovery. Finally, we discuss the techno-economic feasibility and industrial prospects of microbial triterpenoid production, highlight current regulation and governance in synthetic biology related to triterpenoids biosynthesis, analyze existing limitations, and propose potential solutions to provide insights for future research on the biomanufacturing of triterpenoids.

Hypoxia in the tumor microenvironment (TME) is a hallmark of solid tumors and is tightly associated with the development of chemoresistance and immunosuppression, severely compromising the efficacy of mainstream clinical oncological treatments. Platinum-based metallodrugs, especially oxaliplatin (Oxa), serve as first-line chemotherapeutics in clinical practice. However, their clinical utility is greatly restricted by acquired drug resistance, insufficient tumor accumulation, and weak immunostimulatory capacity. Herein, we synthesize a platinum-ruthenium nanohybrid prodrug (denoted as PR) via self-assembly, which integrates Oxa PR and ruthenium ions for synergistic chemo-/chemodynamic-/immunotherapy of hypoxic tumors. The PR nanohybrid possesses intrinsic multi-enzyme activities (catalase, peroxidase, and glutathione peroxidase), enabling efficient oxygen generation, hydroxyl radical production, and glutathione depletion. These cascading events enhance chemosensitivity, trigger robust immunogenic cell death, and activate the cGAS-STING signaling pathway. Furthermore, nanocatalytic modulation of the hypoxic TME alleviates hypoxia-driven immunosuppression, downregulates PD-L1 expression on cancer cells, and reinforces antitumor immune responses. In vitro and in vivo investigations demonstrate that PR nanohybrid exhibits superior anticancer efficacy over free Oxa and displays promising potential in combination with PD-1 blockade therapy. These findings highlight the PR nanohybrid as a versatile TME-modulating platform for hypoxic tumor treatment, offering a novel strategy to advance platinum-based combination cancer therapy.

Dietary protein is a key regulator of metabolic health in humans and rodents. Many of the benefits of protein restriction are mediated by reduced intake of dietary branched-chain amino acids (leucine, valine and isoleucine) and restriction of the branched-chain amino acids is sufficient to extend healthspan and lifespan in mice. Here we find that valine restriction (Val-R) improves metabolic health in C57BL/6J mice, promotes leanness and glycemic control across ages, and reduces frailty, cancer prevalence and senescent cell burden in both sexes while increasing median male lifespan by 23%. Assessing gene relationships across tissues, we identified a liver gene module enriched in mitochondrial pathways and increased mitochondrial respiration in Val-R-fed male mice. Our results demonstrate that Val-R improves multiple aspects of healthspan in mice of both sexes, extends lifespan in male mice and suggests that interventions that mimic Val-R may have translational potential for aging and age-related diseases.

Gastro-entero-pancreatic neuroendocrine tumors (GEP-NETs) are a rare subset of cancers with increasing incidence. Due to their slow growth and lack of targetable mutations, the identification of effective treatments remains limited. One reason behind this stagnation is the lack of applicable, accurate study models. One solution is patient tumor organoids (PTOs) that maintain tumor characteristics and can scale for high throughput assays. In this study, PTOs were generated from 35 tumors of pancreatic, small intestinal, and gastric origin, obtained from 17 patients. Important subtypes including hormone functional and MEN1/VHL mutant GEP-NETs are represented, with each demonstrating growth in culture while maintaining GEP-NET immunohistochemistry and genomic characteristics. Half of G2/G3 tumors (10 of 20) could be cultured past passage 6, whereas G1 tumors (n = 15) were capable of growth until passage 4. Therapeutic targeting of the PTOs displayed both tissue-origin and grade-based response to standard of care and investigational therapies while maintaining patient tumor sensitivity and resistance. Last, a successful PTO xenograft model was developed from one PTO line. This study describes GEP-NET organoid development that demonstrates feasibility for expansion, enabling their use for translational investigations.

Chordomas are rare cancers that arise along the axial skeleton. Alterations in metabolism are a hallmark of cancer, and we sought to identify metabolic vulnerabilities in chordoma. We discovered that the tricarboxylic acid (TCA)-related enzyme isocitrate dehydrogenase-1 (IDH1) was expressed highly in bulk and single-cell patient-derived chordomas and was associated with worse survival outcomes. IDH1 catalyzes the conversion of isocitrate and nicotinamide adenine dinucleotide phosphate (NADP+) to alpha-ketoglutarate (⍺-KG) and NADPH. This critical reaction influences TCA cycle metabolism, regulates epigenetic pathways, and affects redox balance. Both IDH1 knockdown and treatment with an inhibitor targeting IDH1 were toxic to chordoma cells. An integrated analysis of the transcriptomic, chromatin, and metabolomic responses on IDH1 inhibition converged on deregulated glutathione metabolism. IDH1 inhibition was associated with increased expression and enrichment of activating H3K27ac at NRF2 (nuclear factor erythroid 2-related factor 2) signature genes including those in the glutathione biosynthetic pathway. This was accompanied by reduction of both NADPH/NADP+ and reduced/oxidized glutathione (GSH/GSSG) ratios. Importantly, IDH1 inhibitor-driven toxicity was rescued via media supplementation with the antioxidant N-acetylcysteine, suggesting that IDH1 inhibition in chordomas creates a redox-dependent metabolic vulnerability. Finally, IDH1 inhibitor treatment reduced tumor growth in two independent chordoma mouse xenograft models. Our findings suggest a potential therapeutic avenue for further exploration in chordoma.

Cancer stem-like cells contribute to innate tumour immunoresistance and an immunosuppressive tumour microenvironment, leading to poor responses to immune checkpoint inhibitors. Chemotherapeutic agents can elicit tumour immunogenicity by inducing immunogenic cell death to reinforce the therapeutic efficacies of immune checkpoint inhibitors, but suffer from inefficient immunogenic cell death activation in highly resistant cancer stem-like cells. Here we report an immunostimulatory lyotropic liquid-crystal-based lipogel for localized co-delivery of all-trans retinoic acid, a differentiation-inducing drug, and doxorubicin, an immunogenic-cell-death-inducing chemotherapeutic agent with distinct release kinetics. The lipogel is tailored to release the combinatorial drugs in a differential and sustained manner, which fulfils the requirement for enhanced drug synergism in promoting the immunogenic cell death of cancer stem-like cells. Local implantation of the immunostimulatory lipogel elicits an antitumour immune response that is further augmented by an immune checkpoint inhibitor to suppress tumour growth and metastasis, as well as to prevent post-surgical recurrence in murine models of high-stemness tumours.

Basophils, the least abundant leukocytes, are increasingly recognised as potent immunomodulators. Upon activation, they rapidly release preformed granule-associated mediators including histamine and lipid mediators such as LTC4, while cytokine production occurs over a longer timescale, contributing to downstream immune responses. Basophil activation is driven by IgE-dependent FcεRI crosslinking or IL-3, complement and Toll-like receptor (TLR) pathways, enabling diverse effector responses, including type I hypersensitivity to allergens. Through IL-4 and IL-13 secretion, basophils bridge innate and adaptive immunity, promoting Th2 differentiation, B cell activation and recruiting eosinophils. Beyond allergy, basophils contribute to inflammation, angiogenesis, cancer immune surveillance and tumour progression. Basophils participate in IgE-dependent type I hypersensitivity to small-molecule drugs, chemotherapeutics and monoclonal antibodies, becoming activated when drugs cross-link specific IgE-FcεRI complexes, driving rapid mediator release and amplifying Th2-skewed inflammation. The basophil activation test (BAT), which quantifies activation markers, such as CD63 and CD203c, provides an ex vivo functional measurement of these responses and serves as a biomarker of IgE-mediated hypersensitivity. In oncology, BAT is applied to predict and monitor hypersensitivity reactions, drug desensitisation and support IgE-based therapeutics development. Collectively, basophils are emerging as pivotal immune regulators and BAT presents a mechanistic, clinically-actionable tool for advancing precision medicine in AllergoOncology.

Bromodomain-containing protein 4 (BRD4) is an important therapeutic target for anticancer, antiviral, and anti-inflammatory responses. Although its role in transcription, chromatin dynamics, and epigenetic programs is well recognized, recent studies have expanded the functions of BRD4 and its related bromodomain and extra-terminal (BET) family members (BRD2, BRD3, and BRDT) to other molecular and biological processes. How a universal epigenetic regulator is tailored for context-specific gene/pathway regulation is mechanistically intriguing. In this review, we highlight the importance of protein isoforms and posttranslational modifications, particularly phosphorylation, in generating the protein diversity necessary for selective factor recruitment and context-dependent regulation. Three BRD4 protein isoforms have been identified, including the universally expressed BRD4-L and BRD4-S(a) (simplified as BRD4-S), representing the long and short isoform a, and cell-/stress-specific short isoform b, BRD4-S(b). A phospho-switch mechanism controlling the open/closed state of the bromodomain and BRD4 interaction with partner proteins will also be discussed. Mechanistic understanding of BET protein action has led not only to the development of diverse bromodomain-binding compounds currently used in clinical trials but also to the discovery of a new class of small-molecule inhibitors targeting an intrinsically disordered region (IDR) of phospho-BRD4 to alter specific protein-protein interaction (PPI) networks without globally perturbing transcription programs and chromatin landscapes, thus significantly reducing off-target effects and providing new directions for therapeutic drug development.

Autoimmune uveitis is a vision-threatening inflammatory disorder driven by dysregulated T helper 17 (Th17) responses, yet therapeutic strategies targeting Th17 differentiation are lacking. Through transcriptomic screening of an experimental autoimmune uveitis (EAU) model and validation in peripheral blood mononuclear cells from Vogt-Koyanagi-Harada patients, we identified MAP1S (microtubule-associated protein 1S) as a pivotal, conserved regulator. Here, we demonstrate that MAP1S constrains pathogenic Th17 responses and alleviates EAU through a dual mechanism coordinating transcriptional control and autophagic degradation. Mechanistically, MAP1S binds to EGR2 (early growth response 2) and restrains its acetylation at Lys368, thereby suppressing Lcn2 (lipocalin 2) transcription. Besides, MAP1S facilitates autophagosome biogenesis and lysosomal trafficking, promoting the autophagic clearance of LCN2 protein. Notably, MAP1S deficiency enhances EGR2 acetylation, increases Lcn2 transcription, disrupts autophagosome trafficking, impairs LCN2 degradation, and promotes LCN2 accumulation, collectively driving Th17 polarization and exacerbating EAU pathology. Adoptive transfer of cervical lymph node cells from map1s knockout mice reproduced severe disease in wild-type recipients. Moreover, pharmacological activation of MAP1S with spermidine suppressed Th17 responses and alleviated disease severity. Our findings establish MAP1S as a critical node integrating acetylation signaling of EGR2 and autophagic flux to govern LCN2 homeostasis and Th17 pathogenicity, revealing a promising therapeutic target for autoimmune uveitis and potentially other Th17-mediated diseases.Abbreviations: AAV: adeno-associated virus; ACOD1: aconitate decarboxylase 1; AU: autoimmune uveitis; BCL2: B cell leukemia/lymphoma 2; CDLNs: cervical draining lymph nodes; CFA: complete Freund's adjuvant; ChIP: chromatin immunoprecipitation; Co-IP: co-immunoprecipitation; CQ: chloroquine; EAU: experimental autoimmune uveitis; EGR2: early growth response 2; GDF15: growth differentiation factor 15; HDAC4: histone deacetylase 4; HDAC6: histone deacetylase 6; IL17: interleukin 17; IL17f: interleukin 17f; IL22: interleukin 22; K: lysine; KAT2A/GCN5: K(lysine) acetyltransferase 2A; KO: knockout; LCN2: lipocalin 2; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MAP1S: microtubule-associated protein 1S; MS: mass spectrometry; PBMC: peripheral blood mononuclear cell; PCR: polymerase chain rection; PPI: protein-protein interaction; PTX: pertussis toxin; qPCR: quantitative PCR; RT-qPCR: reverse transcription and quantitative real-time RCR; SAA3: serum amyloid A3; SPD: spermidine; Th1 cells: T helper 1 cells; Th17 cells: T helper 17 cells; TF: transcriptional factor; Tregcells: regulatory T cells; VKH disease: Vogt-Koyanagi-Harada disease; WT: wild-type.

Extensive bone defects complicated by infection, malignancy, or metabolic disorders remain a critical clinical challenge, as conventional calcium phosphate bioceramics provide only passive osteoconductive support. The integration of additive manufacturing with external physical stimuli, such as mechanical, piezoelectric, photothermal, magnetothermal, and ultrasonic, has catalyzed a paradigm shift from static scaffolding to responsive therapeutic platforms. This review examines how advanced techniques, including digital light processing (DLP), direct ink writing (DIW), and two-photon lithography (TPL), enable precise architectural programming of porosity, topology, and compositional gradients, establishing the physicochemical foundation for efficient field coupling. We dissect the mechanisms by which field-active bioceramics transduce external stimuli into bioelectrical, thermal, and mechanical cues, activating the mechanotransduction pathway that orchestrates osteogenic differentiation, immunomodulation, angiogenesis, and antibacterial activity. Particular emphasis is placed on multifunctional strategies, including tumor ablation-to-regeneration transitions, antibacterial-to-osteogenic modality switching, and 4D-printed shape memory architectures, alongside emerging self-powered systems harvesting endogenous mechanical energy. By elucidating the synergistic interplay among scaffold structure, material composition, and external field stimulation, this review establishes design principles for next-generation biomaterials that adaptively respond to complex bone-defect microenvironments.

Chemically modified nucleic acids such as antisense oligonucleotides, small interfering RNA and CRISPR guide RNA have emerged as transformative modalities in modern medicine, providing new strategies for the treatment of genetic and metabolic disorders, viral infections and cancer. However, their application is limited by challenges related to intracellular delivery and targeting, advances in which would benefit from a broader range of methods for chemical functionalization. Here we describe a streamlined approach using inexpensive, bench-stable alcohols or nucleosides that enables site-selective modification at the 5' and 3' termini and at internal sites of synthetic oligonucleotides. Our strategy offers a conceptually straightforward solution to this problem through the in situ generation of phosphoramidites on the solid-supported oligonucleotide. The methodology displays high coupling efficiency, high functional-group compatibility with azides, amines and thiols, and efficient recovery of excess alcohols/monomers, thus offering a robust platform for advancing and diversifying therapeutic oligonucleotide modification.

Two live-attenuated vaccines, LMA and LMP, were evaluated alone or in combination with a trivalent adenoviral vector-based vaccine (Ad5-YFV) for their protective efficacy against pneumonic plague in wild-type (WT) and interferon-γ (IFN-γ) knockout (KO) mice. LMA and LMP comprise triple deletion mutants of Yersinia pestis CO92, which causes pneumonic plague, and Ad5-YFV incorporates three protective plague immunogens. Protection of 80 to 100% was observed in vaccinated mice when challenged with highly lethal intranasal doses of parental Y. pestis CO92. All vaccinated mice generated robust humoral and cellular immune responses. Immunized WT mice generated overall greater antibody responses in both serum and bronchoalveolar lavage fluid with higher percentages of polyfunctional T cell populations. Vaccinated IFN-γ KO mice displayed better B cell activity in germinal centers with higher percentages of activated antigen-specific and memory T cells. Superior lung immunity and recall immune responses were observed in both WT and IFN-γ KO mice immunized using a prime-pull vaccine strategy, which also provided full protection against pneumonic plague to mice lacking IFN-α, IFN-β, and IFN-γ receptors. Depletion of IFN-γ or tumor necrosis factor-α from immunized WT mice before and during infection did not reduce protection against pulmonary Y. pestis CO92 challenge. These data suggest that IFN-γ may not be required for protection against pneumonic plague by these vaccines. Combining live-attenuated and adenovirus-based vaccines resulted in augmentation of systemic and local immune responses, which could be beneficial in providing long-lasting immunity against pneumonic plague.

Human ornithine aminotransferase (hOAT), a pyridoxal 5'-phosphate (PLP)-dependent enzyme, plays a central role in glutamine, proline, and polyamine metabolism and is increasingly recognized as a metabolic vulnerability in multiple cancers. Previously, we established a second deprotonation strategy to achieve efficient mechanism-based inactivation of hOAT over closely related aminotransferases. Building on this concept, we report the rational design, synthesis, and mechanistic investigation of cyclopentene-based γ-aminobutyric acid analogues bearing alkyne or nitrile warheads as potent hOAT inactivators. These compounds undergo enzyme-catalyzed γ-deprotonation to form ketimine intermediates, priming for a subsequent tautomerization event that leads to irreversible inhibition. Inhibitory activity evaluation revealed pronounced stereochemical effects on binding affinity and partition ratio, with one nitrile analogue (4b) exhibiting an exceptional inactivation efficiency (kinact/KI = 111.8 mM-1·min-1) and ∼400-fold selectivity for hOAT over γ-aminobutyric acid aminotransferase. Intact protein mass spectrometry and X-ray crystallography demonstrated that alkyne-containing analogues form covalent adducts with hOAT, whereas nitrile-containing analogues generate noncovalent but tight-binding species. Kinetic isotope effect studies identified γ-deprotonation as the rate-determining step, and a complementary small-molecule mass and computational study elucidated the inactivation and turnover pathways. Collectively, these results expand the mechanistic repertoire of PLP-dependent enzyme inactivation and provide a generalizable framework for designing highly selective mechanism-based inactivators.

Integral membrane proteins face unique folding challenges in the endoplasmic reticulum (ER) due to their hydrophobic transmembrane domains and complex topologies. Membrane protein misfolding disrupts ER homeostasis, contributing to diseases such as neurodegeneration, metabolic disorders, cystic fibrosis, and cancers. ER-associated degradation (ERAD) is a protein quality control pathway that detects, ubiquitinates, and retrotranslocates terminally misfolded proteins for proteasomal degradation in the cytosol. Recognition of membrane substrates involves multiple determinants, including topological lesions, exposed hydrophobic patches, and sequence-specific degrons. Recent advances in cryo-EM, functional genomics, and biochemical reconstitution studies have revealed new insights into the mechanistic actions of ERAD. This review addresses the current understanding of ERAD machinery dedicated to targeting misfolded membrane proteins, highlights emerging mechanistic models, and discusses how ERAD drives pathology. Understanding these principles will inform strategies for targeted manipulation of ER membrane protein quality control in health and disease.