胆胰外科 · 本周文献汇编

Obesity is a risk factor for pancreatic cancer, but mechanisms remain unclear. We investigated how anthropometric traits, individually and combined, relate to pancreatic cancer risk and whether associations are mediated by metabolic biomarkers. We analysed 462,300 adults (40-69 years) in the UK Biobank. Principal component analysis derived three body shape phenotypes combining body mass index (BMI), height, weight, waist and hip circumference, and waist-to-hip ratio (WHR). Mediation was assessed using four-way decomposition. Over a median follow-up of 10.9 years, 1115 pancreatic cancer cases occurred. Each one-standard-deviation (SD) increase in BMI or WHR was associated with a higher incidence of pancreatic cancer, with hazard ratios (HRs) of 1.20 (confidence interval, CI: 1.12-1.28) and 1.24 (CI: 1.14-1.36), respectively. Body shape characterizing overall obesity showed a similar association (HR = 1.20; CI: 1.12-1.28 per 1-SD), with glucose and HbA1c accounting for mediated proportions (mediated interaction + pure indirect effect) of 12.2% (CI: 3.4-21.0%) and 15.0% (CI: 5.7-24.2%), respectively. For BMI, glucose accounted for 15.9% (CI: 2.8-28.9%) and HbA1c for 20.0% (CI: 6.3-33.7%) of the association. Glucose and HbA1c mediate a large proportion of the obesity-pancreatic cancer association, highlighting the important role of glycemic control in obesity-related pancreatic carcinogenesis and targeted interventions in at-risk populations.

KRASG12D is the predominant oncogenic driver in pancreatic ductal adenocarcinoma (PDAC). While most investigational KRAS-G12D inhibitors are oral small molecules limited by gastrointestinal toxicities and suboptimal tumor exposure, HRS-4642 is a new, high‑affinity, noncovalent KRAS-G12D inhibitor. Formulated as a liposomal nanoparticle for intravenous administration, it is designed to enhance tumor accumulation and prolong the duration of target inhibition. This phase 1b/2 study evaluated HRS-4642 in combination with nab-paclitaxel and gemcitabine (AG) in patients with advanced KRASG12D-mutant PDAC. As of 5 December 2025, 68 patients were screened and 31 (1 previously treated patient and 30 treatment-naive patients) were enrolled and treated. In the phase 1b portion, no dose-limiting toxicities were observed, and the starting dose (500 mg on day 1 and 1,200 mg on day 8, every 3 weeks) was selected as the recommended phase 2 dose. In the phase 2 portion, with a median follow-up of 12.3 months (95% confidence interval (CI) = 12.2-13.0), the primary endpoint was met-the confirmed objective response rate in 30 treatment-naive patients was 63.3% (95% CI = 43.9-80.1). Grade ≥3 treatment-related adverse events (TRAEs) occurred in 90.3% patients, primarily hematologic toxicities consistent with AG chemotherapy. No TRAEs led to treatment discontinuation or death. In conclusion, HRS-4642 combined with AG demonstrates promising antitumor activity and a manageable safety profile in advanced KRASG12D-mutant PDAC, supporting further investigation. Clinicaltrials.gov registration: NCT06520488 .

Pain is a major concern in patients with advanced pancreatic cancer, significantly affecting quality of life, treatment tolerance, and end-of-life experience. However, data on its prevalence, course, and determinants remain limited, particularly regarding the dynamics of pain as a primary outcome in real-world patient populations. This nationwide cohort study, conducted within the PACAP-PROM registry (2016-2023), included patients with locally advanced (LAPC) or metastatic pancreatic cancer (mPC). Pain was assessed using 5 items from the EORTC QLQ-C30, PAN26, and EQ-5D-5L questionnaires, with the primary outcome being pain prevalence at baseline and comparisons between LAPC and mPC. Secondary outcomes included pain trajectories over 3 to 18 months and the association between chemotherapy and pain, evaluated using mixed-effects logistic regression. Overall, 794 patients were included, of whom 648 (82%) received cancer therapy, and 146 (18%) received best supportive care (BSC). At baseline, 54% to 78% of patients reported pain, with no differences between LAPC and mPC. Although pain prevalence decreased at 3 months, variable patterns developed thereafter. Patients receiving BSC reported more pain (57% vs 38%, P = 0.029) and greater pain-related interference with daily activities (53% vs 38%, P = 0.024). Chemotherapy was independently associated with lower odds of higher pain severity categories for current pain (OR 0.51; 95% CI 0.37-0.70) and pain interference (OR 0.44; 95% CI 0.31-0.63). In conclusion, pain prevalence exceeds 50% in patients with advanced pancreatic cancer and remains a key concern that can be positively influenced by chemotherapy. These findings emphasize the need for routine pain assessment and individualized symptom management from diagnosis onward.

The efficacy of pharmacological glycogen synthase kinase-3β (GSK3β) inhibition is currently being investigated in unselected cohorts of metastatic pancreatic ductal adenocarcinoma (PDAC). Here, we sought to determine the clinical significance of nuclear GSK3β accumulation in patients with resectable PDAC. This study aimed to explore the therapeutic potential and underlying mechanisms of GSK3β pathway disruption in PDAC with enriched nuclear GSK3β levels. We investigated the activation and function of GSK3β and its downstream transcription factor NFATc1 in tumour recurrence, growth and resistance using human PDAC tissues, patient-derived organoids and tumour cells, PDAC explants, cell lines and murine models. GSK3β signalling was disrupted using genetic and pharmacological approaches. Live-cell imaging, proliferation, homologous recombination (HR) repair and comet assays, messenger RNA sequencing and chromatin immunoprecipitation were used to explore GSK3β-NFATc1 signalling-mediated target gene regulation in DNA repair, growth and resistance. Nuclear GSK3β accumulates in a subset of resected PDAC and promotes proliferation and DNA repair through NFATc1. The GSK3βhigh/NFATc1high subtype accounts for 14% of resected PDAC and is associated with rapid tumour recurrence and poor survival. The GSK3β-NFATc1 signalling pathway contributes to cisplatin resistance by inducing BRCA genes transcription, which facilitates HR-mediated DNA double-strand breaks (DSBs) repair. Disruption of the GSK3β-NFATc1 axis impairs HR-driven DSB repair, increasing cisplatin sensitivity in vitro and in preclinical PDAC models. We have identified a highly aggressive GSK3βhigh/NFATc1high subtype that predicts early recurrence, poor survival and cisplatin resistance in PDAC. This subtype reveals new treatment vulnerabilities suggesting that patients with PDAC may benefit from stratification-based tailored treatment strategies.

Cholesterol metabolic reprogramming plays a key role in pancreatic cancer progression by regulating the tumor immune microenvironment (TIME) and influencing responses to immunotherapy. Pancreatic intraepithelial neoplasia (PanIN) is the most common precursor lesion of pancreatic ductal adenocarcinoma (PDAC), a malignancy that commonly occurs in individuals with high BMI and type 2 diabetes mellitus (T2DM). Key metabolic markers (SOAT1 and SREBP2), the tumor suppressor P53, and the immune marker CD8 are implicated in PanIN initiation, PDAC progression, and subsequent metastasis, but their associations-particularly for those that are potentially targetable-remain poorly defined. Using publicly available PDAC datasets from the GEO database, we conducted in silico analyses to investigate the genes of interest. Additionally, we used laser capture microdissection to isolate specific cells from pancreatic cancer tissues obtained from patients. We performed translational research using 17 tumor samples, as well as 17 microdissected PanIN lesions and matched normal tissues. Protein expression analysis was further conducted on a larger cohort, including 65 PanIN lesions and 157 tumor tissues from PDAC patients. Our data identify a dual-direction expression pattern in obese and diabetic patients, with elevated levels of SOAT1, SREBP2, and P53, and reduced CD8 expression, suggesting a coordinated metabolic and immunological alteration in pancreatic cancer. High BMI and T2DM, recognized as independent risk factors for PDAC, are significantly associated with dysregulation of key markers involved in the mevalonate pathway and the TIME, potentially contributing to PanIN and PDAC progression. These findings warrant further longitudinal and mechanistic studies to clarify whether metabolic modulation may influence PDAC risk or clinical outcomes.

Nano-flow cytometry (nFCM) is a single-particle analytical technique for the detection, sizing, and phenotyping of extracellular vesicles (EVs). However, nFCM analysis of small-sized EVs (<100 nm) or those with low-abundance antigen markers for immunostaining remains a major challenge. In this work, we developed a primer exchange reaction-based dual-signal amplified nano-flow cytometry (PER-DSA nFCM) method for the specific detection of MUC1-positive EVs and the phenotyping of exosomes derived from different pancreatic cancer cell lines. The mechanism relies on the specific binding of a DNA hairpin to the MUC1 protein on the EVs surface, followed by the two-dimensional expansion of the opened hairpin via primer exchange reaction to enlarge individual EVs. The enlarged EVs with extended DNA polymers can hybridize with multiple molecular beacons, emitting strong fluorescence, which can be detected by nFCM through the amplified laser side-scattering signal and fluorescence signal. Conventional fluorescence measurement can be also employed for bulk concentration quantification to the expanded EVs. Using PER-DSA nFCM, we successfully distinguished exosomes secreted from pancreatic cancer cell lines HPAC and BxPC-3, which exhibit different surface MUC1 expression levels. Bulk analysis of HPAC exosomes via post-amplification solution fluorescence measurement yielded a good linear quantitative response to concentration of EVs in the range of 5 × 105-7.5 × 106 particles μL-1, with a detection limit of 1.25 × 103 particles μL-1. Applying this assay in plasma biopsy for diagnosing pancreatic cancer, the receiver operating characteristic curve presented a sensitivity of 77.8% and a specificity of 93.3% at the best cutoff, with an area under the curve of 0.844. The proposed method is simple, rapid, and sensitive for the analysis of MUC1-positive EVs, and can be easily modified to detect EVs expressing other surface protein markers, holding potential for clinical applications.

Pancreatic ductal adenocarcinoma has a unique tumor microbiome, and the depletion of gut bacteria or fungi using antibiotic/antifungal cocktails has been shown to decrease pancreatic tumor burden in mice. However, functional studies evaluating the role of tumor-associated microbes are few due to the limited availability of clinically relevant microbiota. Here, we describe in detail an effective workflow for the isolation of bacteria and fungi from the duodenum and tumor of pancreatic cancer patients, specifically optimized for cryopreserved, low biomass samples. Using this workflow we also isolated microbiota from normal pancreatic tissue and duodenum from organ donors, and we confirmed the presence of bacteria and fungi isolated from tissue samples with 16S and ITS sequencing analysis. Isolation and sequencing results show distinct similarities between the pancreatic and duodenal microbiomes and highlight unique bacterial strains that survive in the tumor microenvironment. As a proof of concept, we characterized a select Klebsiella oxytoca strain (UMKO1) isolated from a pancreatic tumor, using whole genome sequencing, metabolomics, and ex- vivo tumor cultures to determine its potential impact on the pancreatic tumor microenvironment. In summary, this optimized workflow allows for the isolation of a variety of bacteria and fungi from low biomass, cryopreserved pancreatic and duodenal tissues, which can then be used for functional studies characterizing clinically relevant tumor-associated microbiota.

Solute carrier family 1 member 1 (SLC1A1, also known as excitatory amino acid transporter 3) is a member of the EAAT family of sodium-dependent glutamate carriers. Ferroptosis is a nonapoptotic cell death that is caused by phospholipid peroxidation promoted by radical reactions involving iron. Here, we identify SLC1A1 as a driver of ferroptosis resistance in PDAC cells and delineate the cause of its upregulation. SLC1A1 was overexpressed in PDAC tumors and correlated with poorer prognosis. Knockdown of SLC1A1 reduced cystine uptake by inhibiting glutamate transport, leading to decreased cysteine, GSH, and glutathione peroxidase 4 (GPX4) content, and the production of ROS and lipid peroxidation. Knockdown of SLC1A1 enhanced the ferroptosis sensitivity of PDAC cells and promoted ferroptosis inducer RSL3-induced ferroptosis and tumor suppression. Long noncoding RNA RASSF1-AS1 bound to CCCTC-binding factor (CTCF) protein, thereby inhibiting CTCF-mediated SLC1A1 transcription. Overexpression of RASSF1-AS1 enhanced the ferroptosis sensitivity of PDAC by blocking glutamate transport in vitro and in vivo, which was reversed by restoring CTCF/SLC1A1 signaling. Taken together, RASSF1-AS1 enhances the ferroptosis sensitivity of PDAC by inhibiting the CTCF/SLC1A1 axis-mediated intracellular transport of glutamate.

Epithelial splicing regulatory protein 1 (ESRP1) is a key regulator of epithelial-mesenchymal transition (EMT) and cancer progression, including in pancreatic ductal adenocarcinoma (PDAC). We show that ESRP1-mediated isoform switching of DNA methyltransferase 3B (DNMT3B) correlates with the metastatic potential of pancreatic cancer cells. The expression of DNMT3B splicing variants was closely linked to ESRP1 level. Ectopic expression of mesenchymal DNMT3B isoforms 3 and 7, lacking part of the methyltransferase catalytic domain, significantly enhanced lung metastasis and tumor growth, while epithelial DNMT3B isoforms 1 and 2, with an intact catalytic domain, suppressed these effects in mouse models. Differential expression of DNMT3B isoforms also correlated with overall survival in PDAC patients. Deletion of exons 21-22, differentially spliced region between epithelial and mesenchymal isoforms, increased cell proliferation and migration, similar to mesenchymal DNMT3B. RNA sequencing revealed that mesenchymal DNMT3B isoforms 3 and 7 were associated with aggressive tumor phenotypes. Among differentially expressed target genes of DNMT3B isoforms, Fibulin 1 (FBLN1), a key regulator of the tumor microenvironment, regulates cancer cell migration and EMT. Taken together, our results highlight the central role of ESRP1-mediated DNMT3B isoform switching in pancreatic cancer progression and metastasis and suggest that DNMT3B isoforms may serve as potential biomarkers for this disease.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a pronounced Warburg effect and high lactate levels. While lysine lactylation (Kla) is an emerging post-translational modification, its role in regulating the S100A11/ANXA2 complex-a critical driver of membrane repair and metastasis-remains unexplored. In this study, we identified that S100A11 (K3, K55) and ANXA2 (K49) are significantly lactylated in PDAC tissues. Lactate-induced lactylation of S100A11 at K3 and K55 inhibits FBXW11-mediated ubiquitination and proteasomal degradation, thereby stabilizing the protein. This stabilized S100A11 subsequently shields ANXA2 from TRIM21-mediated degradation. Reciprocally, lactylation of ANXA2 at K49 enhances its capacity to recruit S100A11 to the plasma membrane. Functional assays demonstrated that this lactylation-driven interdependent regulatory axis promotes PDAC cell migration, invasion, and metastasis both in vitro and in vivo. Clinical analysis of TCGA data revealed that low expression of both S100A11 and ANXA2 predicts significantly improved progression-free and overall survival in PDAC patients. In summary, our findings establish a novel link between metabolic reprogramming and the post-translational regulation of the membrane repair machinery. Lactylation of the S100A11/ANXA2 axis is a key driver of PDAC progression and serves as a promising prognostic biomarker and potential therapeutic target.

Pancreatic cancer remains one of the most lethal malignancies, characterized by a highly immunosuppressive tumor microenvironment (TME) and a dense desmoplastic stroma, representing a significant challenge to immunotherapy application. This systematic review evaluates cytokine-based immunotherapy as a strategy to overcome these barriers. Following PRISMA guidelines, we searched PubMed, SCOPUS, and Web of Science for original preclinical studies published between 2020 and 2025 that evaluated the administration or inhibition of cytokines in monotherapy or combined therapy with checkpoint inhibitors and chemotherapy. Risk of bias was assessed with the QUIN (in vitro) and SYRCLE (in vivo) tools. A total of 46 studies were included, revealing two primary therapeutic approaches: blocking chronic immunosuppressive factors and administering immunostimulatory cytokines. Targeting cytokines such as TGF-β and IL-6 induced high stromal remodeling and fibrosis reduction, thereby facilitating effector cell infiltration. Conversely, the delivery of pro-inflammatory cytokines (e.g. IL-2, IL-12 or IL-15) demonstrated significant potential in enhancing T-cell persistence and preventing exhaustion. Cytokine modulation was rarely sufficient as a monotherapy but produced robust antitumor responses when combined with chemotherapy or immune checkpoint inhibitors (anti-PD-1). Evidence was limited by the predominant use of subcutaneous and xenograft models that incompletely reproduce the human pancreatic stroma and by an unclear-to-high risk of bias among in vivo studies. In conclusion, manipulating the cytokine environment can convert the "cold" pancreatic tumor microenvironment into a "hot" state. Although integrating these strategies into multimodal regimens shows promise, rigorous clinical validation is essential to determine their true efficacy and safety in humans.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by high malignancy and poor prognosis. However, chemotherapy remains the cornerstone of its treatment. Through transcriptomic analysis of gemcitabine-treated PDAC cells, we identified TRDMT1, rather than other known 5-methylcytosine (m5C) methyltransferases, as a principal regulator of mRNA m5C methylation. TRDMT1 deficiency attenuated the malignant phenotype of PDAC cells and increased their sensitivity to gemcitabine, 5-fluorouracil, oxaliplatin, and irinotecan. Furthermore, deletion of TRDMT1 impaired the recruitment of homologous recombination-related proteins, including BRCA1, RAD51, and RAD52. Analysis of mRNA m5C modification sequencing in TRDMT1-deficient PDAC cells demonstrated that IRS2 exhibited significantly reduced m5C methylation levels and decreased expression. The IRS2/PI3K/AKT axis was identified as the potential downstream signaling pathway through which TRDMT1 mediated the chemotherapeutic response. Survival data from patients with PDAC, and findings from TRDMT1-deficient mouse models, further supported the role of TRDMT1 in promoting chemoresistance. Collectively, these findings indicate that TRDMT1-mediated mRNA m5C methylation is essential for homologous recombination repair in PDAC and suggest that TRDMT1 may represent a potential therapeutic target for overcoming chemoresistance.

Cancer stem cells (CSCs) play a pivotal role in the initiation, metastasis, and recurrence of pancreatic tumors, significantly contributing to drug resistance and reducing the efficacy of traditional chemotherapy. To address these challenges, we developed an amphipathic self-assembled nano-prodrug to co-deliver all-trans retinoic acid (ATRA), gemcitabine (GEM), and ferroptosis inducer (RSL3) for the combined therapy of pancreatic ductal adenocarcinoma (PDAC). The amphipathic prodrug (ATRA-GEM) was synthesized by conjugating ATRA and GEM via an ester bond, enabling good self-assembly and the simultaneous loading of the ferroptosis inducer RSL3. In tumor microenvironment, this combinational nano-prodrug disassembles in response to high level of esterase, triggering the controlled release of ATRA, RSL3, and GEM. The released drugs exhibited a combinational therapeutic effect in vitro by combining apoptosis, ferroptosis, and differentiation therapy. For in vivo application, the nano-prodrug was modified with DSPE-PEG-RGD to enhance tumor-targeting capability and prolong blood circulation. This nano-prodrug demonstrated significant anti-PDAC efficacy and good biosafety in both cell-derived xenograft and patient-derived xenograft models. STATEMENT OF SIGNIFICANCE: Pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest cancers due to drug resistance and tumor relapse driven by cancer stem cells (CSCs). Here, we report an amphipathic self-assembled nano-prodrug that simultaneously delivers a differentiation agent (ATRA), a chemotherapeutic (gemcitabine), and a ferroptosis inducer (RSL3). Unlike existing therapies that target only one tumor population, this strategy integrates CSC differentiation, ferroptosis-induced cell death, and chemotherapy into a single platform, enabling coordinated elimination of both CSCs and bulk tumor cells. This nano-prodrug also features high drug-loading capacity and tumor-targeted, stimulus-responsive release. This work provides a new strategy for overcoming resistance and achieving more durable therapeutic outcomes in PDAC, with broad implications for treating other refractory cancers.

The hallmarks of cancer and enabling characteristics are widely regarded as the key features that best define malignant disease. In a process as intricate as the development of cancer, a simple definition or stepwise series of biological events does little to encapsulate the insurmountable complexity that is this disease. Indeed, over the course of the last 25 years alone, there have been four renditions of the "Hallmarks of Cancer", each attempting to encapsulate the mechanisms involved in the development of cancer by providing a list of strict criteria that must be met in order to define the disease. With each of these four sets of principles, modern research has caused the ensuing recapitulation to be revised, and without exception, to expand. In fact, what originally began as six hallmarks of cancer, has become a list of parameters that is fourteen strong, and which aim to summarize the functional capabilities that, when acquired by human cells, provides them with the ability to form a malignant, life-threatening tumour. While at present these hallmark capabilities have largely been discussed in the context of all cancers, this review discusses each in the specific context of pancreatic cancer (PC). PC has the worst prognosis of all cancers globally, with late-stage diagnoses, aggressive tumour microenvironments, and limited treatment options contributing to the current 5-year survival rate of 13%. Here, we highlight PC-specific mechanisms underpinning each hallmark of cancer to provide a comprehensive review of the current state of knowledge around this complex disease, with the aim of aiding understanding and facilitating more meaningful research avenues.

Methionine dependence represents a well-known metabolic vulnerability in cancer. Despite promising preclinical results, methionine restriction is impaired by the presence of methionine-independent cancer cells. After confirming both inter- and intra-tumoral heterogeneity in methionine dependence, we demonstrate that "methionine-independent" cells are rather "methionine self-sufficient," relying on the vitamin B12 (B12)-dependent methionine synthase (MTR) to sustain growth without exogenous methionine, which renders them highly vulnerable to B12 deprivation. Dual methionine and B12 deprivation produced synergistic cytotoxicity, inhibiting proliferation and inducing apoptosis across multiple cancer types and primary tumor cells, while sparing fibroblasts. This synergy persisted under moderate nutrient restriction, supporting translational potential. Moreover, dual therapy prevented the adaptive metabolic shift seen with methionine deprivation alone, avoiding rebound proliferation and resistance. In vivo, a methionine-restricted diet plus a synthesized B12 antagonist significantly suppressed growth of methionine-independent pancreatic xenografts without hematologic toxicity. These findings uncover a selective, synergistic anticancer strategy targeting methionine self-sufficiency.

Nature killer (NK) cell plays a critical role in cancer immunosurveillance and is considered a potent immunotherapeutic tool for many cancers, including pancreatic ductal adenocarcinomas (PDACs). Increasing evidence suggests that cancer occurs with systemic immune perturbations. However, the effects of PDAC tumor burden on systemic NK cells remain poorly understood. PDAC tumor-bearing mice display decreased frequency and dysfunction of NKs the spleens. We identified an increase in Gr-1+ myeloid cells within the spleens, which negatively impacts both endogenous and adoptively transferred NK cell frequency and function. Apolipoprotein E (ApoE), a lipid metabolism regulator, is upregulated in Gr-1+ myeloid cells of tumor-bearing mice, promoting lipid oxidation and reactive oxygen species (ROS) generation. Genetic knockout of Apoe in Gr-1+ myeloid cells abrogate their suppressive effects on NK cell function. Furthermore, treatment with lipid metabolism inhibitors restores endogenous and adoptively transferred NKs' effector function in the spleens and tumor microenvironment. These studies underscore the importance of understanding preexisting systemic alterations in PDAC patients before applying NK cell-based immunotherapies.

The development of various cancers is intricately linked with the human microbiome. Recent studies have highlighted a substantial association between pancreatic cancer and gut microbiota. However, the specific roles and underlying regulatory mechanisms of individual gut microbial species in pancreatic cancer progression remain poorly understood. Enterococcus faecalis, a common member of the human commensal microbiota, has been found to be enriched in the tumor tissues of pancreatic cancer patients. However, its functional contribution has not been clearly defined. In this study, we established a co-culture system involving E. faecalis and pancreatic cancer cells. Our results show that E. faecalis promoted the proliferation, migration, and invasion of pancreatic cancer cells. Following pre-treatment with E. faecalis, the phosphorylation level of epidermal growth factor receptor (EGFR) was markedly elevated. Inhibition of EGFR effectively suppressed the pro-proliferative effects induced by E. faecalis. Further investigation revealed that E. faecalis stimulated the production of reactive oxygen species (ROS) in pancreatic cancer cells. This ROS production might be sensed by Toll-like receptors (TLRs), leading to the activation of the EGFR signaling pathway. When cells were incubated with TLR inhibitors or ROS scavengers, both EGFR expression and its downstream pro-proliferative effects were significantly attenuated. Collectively, this study provides mechanistic insights into how E. faecalis contributes to pancreatic cancer progression and offers new perspectives for the development of diagnostic and therapeutic strategies targeting this microbial signaling pathway.IMPORTANCEA diverse microbiome is closely associated with cancer, as bacterial presence has been detected in the majority of solid tumors. However, the composition, abundance, and functional profiles of the microbiota vary significantly across different tumor types, thereby exerting distinct effects on tumorigenesis and disease progression. Recent studies have shown that pancreatic cancer hosts a variety of bacterial populations, including gut-derived bacteria that may translocate to pancreatic tissue via mesenteric venous or lymphatic drainage pathways. For example, Enterococcus and Enterobacter species have been identified in the cyst fluid of patients with pancreatic cystic neoplasms. Moreover, antibodies against Enterococcus faecalis capsular polysaccharide have been detected in the sera of patients with pancreatitis and pancreatic cancer, and E. faecalis has been observed in pancreatic ducts. Despite these observations, the precise mechanisms through which E. faecalis influences pancreatic cancer remain unclear. Our study demonstrates that E. faecalis promotes pancreatic cancer cell proliferation through the activation of the Toll-like receptor-reactive oxygen species-epidermal growth factor receptor signaling pathway.

Antibody production by B cells has emerged as an important factor in regulating antitumor immunity with both suppressive and promotive roles in cancer. However, the specific effect of antibody deficiency during development of pancreatic ductal adenocarcinoma (PDAC) has not been explored. To address this question, we crossed the well-established KPC mouse model to mice lacking all circulating immunoglobulin (Ig) due to genetic ablation of both Ig secretion and Ig class switching (KPC-μSAID mice). KPC-μSAID mice exhibited a two-fold acceleration in tumor formation, a two-fold reduction in median survival, and increased liver metastases versus KPC-WT control mice. Immunofluorescence analysis of pancreatic tissues from antibody-sufficient KC- and KPC-WT mice showed that IgG was predominantly localized within the extracellular matrix (ECM). Furthermore, in both KC- and KPC-μSAID mice, ECM density and podoplanin+ cancer-associated fibroblasts (CAFs) were significantly reduced. In the KPC-μSAID tumor microenvironment (TME), intratumoral myeloid-derived suppressor cells (MDSC) were also increased, while CD4+ and CD8+ T cells decreased, relative to tumor-bearing KPC-WT mice, with macrophage exhibiting a mixed polarization phenotype. These findings were recapitulated in antibody subclass-deficient, KPC-AID mice, suggesting a potentially novel function of IgG in suppressing PDAC progression by directly or indirectly regulating pancreatic fibrosis and the density of the ECM.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy characterised by remarkable cellular heterogeneity, which emerges early from the interplay of oncogenic KRAS signalling and inflammatory injury. However, the transcriptional, metabolic and functional properties of these pre-malignant cell states that initiate and drive PDAC progression remain elusive. This study aimed to identify and functionally characterise the critical premalignant cell states that arise from this heterogeneity, to define novel biomarkers and targets for early intervention. Public and in-house scRNA-seq data of pancreatic tumour models were analysed to identify key subpopulations in early cellular heterogeneity. Genetic perturbation in KrasG12D-driven models was performed to assess functional impact. Mechanistic studies used TurboID proximity proteomics, epigenetic profiling and metabolic assays. Clinical relevance was validated in human PDAC cohorts. We identified LY6D as a marker of a distinct, gastric-like cell state that emerges early and persists throughout tumourigenesis. The LY6D+ population exhibits conserved stemness and a unique, pan-stage dependency on oxidative phosphorylation (OXPHOS). Genetic ablation of Ly6d specifically impaired the gastric lineage and delayed tumourigenesis, while its overexpression enhanced tumourigenic and metastatic potential. Mechanistically, the glycosylphosphatidylinositol (GPI)-anchored LY6D protein scaffolds a lipid raft-associated kinase network that drives FOSL1-dependent epigenetic-transcriptional reprogramming. In human PDAC, LY6D+ cells harbour stemness and Epithelial-Mesenchymal Transition (EMT) signatures, and high LY6D expression is an independent prognostic marker of poor survival. Our work defines the LY6D+ gastric-like cell state as a key driver linking early pre-malignant heterogeneity to PDAC initiation and progression. LY6D represents a pan-stage therapeutic target and a candidate biomarker for early detection and therapeutic targeting.

The biological significance of forkhead box A1 (FOXA1) in non-steroid-driven malignancies, such as pancreatic ductal adenocarcinoma (PDAC), has garnered increasing recognition. It assumes a pivotal role in regulating critical processes such as PDAC cell lineage, metabolism, and metastasis. However, its regulatory mechanisms remain elusive. Here, we demonstrate that histone deacetylase 5 (HDAC5) mediates the deacetylation of FOXA1 at lysine residue 270 (K270), leading to repression of FOXA1's global chromatin occupancy. In HDAC5-loss PDAC, K270 hyper-acetylated FOXA1 is reprogrammed to the transcription start sites (TSSs) of HIF1α-targeted genes, functioning as a pioneer factor of HIF1α signaling. Additionally, we show that HDAC5 antagonizes LSD1-mediated FOXA1 activation by converging on the dynamic equilibrium of acetylation-methylation transition at K270. Pharmacological inhibition of HIF1α/LSD1 suppresses the growth and progression of HDAC5-deficient PDAC in in vivo and in vitro models. Our study reveals the role of FOXA1 as a pioneer factor of HIF1α in PDAC, providing potential therapeutic strategies for HDAC5-deficient PDAC.

Pancreatic ductal adenocarcinoma (PDAC) exhibits profound therapy resistance driven by lysosome-dependent nutrient recycling, metabolic adaptation, and stress tolerance. Current lysosome targeting agents such as chloroquine (CQ)/hydroxychloroquine (HCQ) show limited efficacy due to transient activity and dose-limiting-toxicities. To overcome these limitations, we developed lysostilbenes, a new class of hybrid small molecules combining the CQ pharmacophore with lysosome-disrupting stilbene analogs. Stilbene pharmacophore is the core structural component of resveratrol. Among the synthesized hybrids, lysostilbene-4 emerged as the lead candidate, demonstrating ~30-40-fold greater cytotoxicity against PDAC cells than parent compounds, while sparing nonmalignant cells. At nanomolar concentrations, lysostilbene-4 induced rapid, irreversible lysosomal membrane permeabilization (LMP), initiating a lysosome mitochondria apoptotic cascade via CTSB (cathepsin B) release, BID cleavage, BAX activation, and caspase-mediated apoptosis. In parallel, it abrogated lysosomal recovery by significantly reducing repair, lysophagy, autophagosome maturation, and uncoupling TFEB-driven transcriptional programs from effective lysosome biogenesis. Reduced TFEB mRNA expression correlated with poor overall-survival and disease-free-survival across multiple cancer patients, with a particularly strong association in pancreatic cancer patients. Using TFEB+/+ and TFEB-/- knockout pancreatic cancer cells we establish that lysostilbene-4 exerts severe cytotoxicity by inducing persistent lysosomal-damage and disrupting autophagosome-lysosome assembly, with vulnerability further amplified in TFEB-deficient cells. This finding underscores TFEB as a key determinant of lysosomal-resilience and a potential predictive biomarker. Importantly, lysostilbene-4 was well tolerated in preclinical mouse-models at supra-therapeutic doses without systemic-toxicity. These findings position lysostilbene-4 as a first-in-class lysosome-targeting therapeutic that enforces sustained lysosomal collapse while compromising adaptive recovery-mechanisms, providing a mechanistically precise and safe strategy against PDAC.Abbreviations: ALG: autophagy-lysosome genes; AMPK: AMP-activated protein kinase; CASM: conjugation of ATG8s to single membranes; CTSB: cathepsin B; LGALS3: galectin 3; LMP: lysosomal membrane permeabilization; LS: lysostilbene; MTOR: mechanistic target of rapamycin kinase; PDAC: pancreatic ductal adenocarcinoma; TCGA: The Cancer Genome Atlas; TFEB: transcription factor EB; ULK1: unc-51 like autophagy activating kinase 1.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest cancers due to late diagnosis, early metastasis and resistance to therapy. Cancer stem cells (CSCs) have been implicated in PDAC aggressiveness and treatment failure. MAP kinase-interacting kinase 1 (MNK1) is overexpressed in PDAC and plays a critical role in tumor progression and CSC maintenance. Here, we show the potential of apMNKQ2, a DNA aptamer targeting MNK1, to therapeutically target CSCs and reduce PDAC tumor burden in patient-derived xenografts (PDXs). PDX cell lines and in vivo mouse models were used to assess the effects of apMNKQ2 on cell viability, apoptosis, cell cycle progression, migration, epithelial-to-mesenchymal transition (EMT) and CSC properties. Functional CSC targeting was validated through clonogenic and self-renewal assays as well as extreme limiting dilution analysis. Systemic administration of free apMNKQ2 was tested for biodistribution, pharmacokinetics, toxicity, and antitumor efficacy at escalating doses. apMNKQ2 downregulated MNK1 and anti-apoptotic proteins (MCL1, XIAP), impaired cell proliferation, induced apoptosis, and disrupted cell cycle progression in PDX PDAC cells. Importantly, apMNKQ2 also inhibited migration, mesenchymal properties and angiogenesis in vitro, and lung colonization in vivo. Notably, apMNKQ2 strongly targeted PDAC CSCs, reducing CD24, CD133, CXCR4 and ALDH expression, clonogenicity and in vivo tumor initiation over 600-fold. Free apMNKQ2 (without transfection agents) entered PDAC cells efficiently, retaining anti-CSC activity. Systemic delivery of free apMNKQ2 accumulated in tumors, was well tolerated up to 400 mg/kg and showed no toxicity. Importantly, 10 mg/kg of apMNKQ2 produced strong antitumor effects in PDX models. Increasing the dose 20-fold enhanced tumor uptake but not efficacy, suggesting a therapeutic plateau at 10 mg/kg. MNK1 plays a central role in PDAC progression and CSC maintenance. apMNKQ2 is a potent anti-MNK1 DNA aptamer with robust preclinical activity, including CSC-targeting and anti-invasive effects. Its low toxicity, systemic bioavailability, and efficacy at low doses support further development as a novel therapeutic strategy for PDAC.

Pancreatic cancer remains a lethal malignancy with limited treatment options. Trophoblast cell surface antigen 2 (TROP2), which is frequently overexpressed in pancreatic tumors and closely associated with poor prognosis, represents a promising therapeutic target. hIMB1636-LDM is a novel antibody-drug conjugate (ADC) developed by our laboratory, comprising the anti-TROP2 antibody hIMB1636 covalently linked to cytotoxic payload lidamycin (LDM) via an uncleavable linker. In this study, we systematically evaluated the antitumor efficacy and underlying mechanism of action of hIMB1636-LDM in pancreatic cancer. The results showed that hIMB1636-LDM exhibited TROP2-dependent binding and internalization, as well as potent cytotoxicity (Half maximal inhibitory concentration (IC50): 0.27-0.5 nM) against pancreatic cancer cells. Mechanistically, beyond inducing cell cycle arrest and apoptosis, it was for the first time found to possess the ability to trigger immunogenic cell death (ICD). In vivo, hIMB1636-LDM at a dose of 0.8 mg/kg significantly suppressed tumor growth in both T3M4 and BxPC3 models, with no obvious toxic reactions observed. hIMB1636-LDM exerts potent antitumor activity through a dual mechanism combining targeted cytotoxicity and ICD induction. Its favorable safety profile, potent efficacy, and unprecedented ICD-inducing property highlight its potential as a promising therapeutic candidate for TROP2-positive pancreatic cancer.

A liposomal CPT-11 formulation was previously developed and approved for pancreatic cancer (Onivyde), but it carries a FDA black-box warning because it exacerbated CPT-11 associated gastrointestinal (GI) toxicities. An alternative liposomal irinotecan formulation, Irinophore C, reduced GI toxicity as judged by a validated animal model of irinotecan mediated early and late onset diarrhea. Irinophore C did not progress to clinical trials, partly because its development coincided with the approval of Onivyde. Nevertheless, an Irinophore C-like formulation remains of interest because of its potential to reduce GI toxicity and its potential to be developed as a combination product. Irinophore C was prepared using liposomes containing entrapped copper (Cu2+). The divalent metal ionophore A23187 was used to exchange entrapped Cu2+ for protons, creating a transmembrane pH gradient. Following addition of CPT-11 to the liposomes, it crossed into the liposome interior to achieve efficient (>95%) liposome association. The results were consistent with previous findings that CPT-11 can be loaded into liposomes with a transmembrane pH gradient (inside acid). The presence of copper, however, imparted unique properties. The resulting formulation retained CPT-11 better than liposomes loaded using a simple transmembrane pH gradient or a pH gradient generated with A23187 added to liposomes containing other divalent metal ions (e.g., Mg2+, Mn2+, Zn2+). The data suggested improved drug retention arose due to Cu2+ binding with CPT-11 and/or with lipids in the inner membrane of the liposomes. It was further suggested that Cu2+ may mitigate GI toxicity. In this study it is argued that if Cu2+ ions confer unique advantages to the liposomal CPT-11 formulation, then the original method used to prepare Irinophore C was not optimal. This is because A23187 significantly reduced the amount of copper retained in the liposomes. Here, we characterize an alternative method for CPT-11 encapsulation; a method that retains Cu2+ within liposomes. In this formulation, A23187 is not required and the low internal pH is maintained with ammonium sulfate. The results showed that CPT-11 can be efficiently loaded into these liposomes, achieving CPT-11 to liposomal-lipid ratios exceeding 0.6 (mol:mol). The resulting formulation exhibits significantly improved drug retention and cryo-transmission electron microscopy analysis suggests the morphology of the new formulation differs from Irinophore C. Efficacy studies in a syngeneic BALB/c CT26 colorectal cancer model demonstrated the therapeutic potential of the new formulation, but highlight a challenge when developing high CPT-11 to liposomal-lipid ratio formulations. Specifically, high drug to liposomal-lipid formulations utilize significantly lower lipid doses that are eliminated rapidly after administration. It is suggested that this new formulation is suitable for development in combination with other therapies, however the rapid elimination of the liposomes administered at low liposomal lipid dosed must be addressed.

The pancreatic tissue is composite and plastic. The epigenetic makeup of different cell types is poorly characterized due to the lack of protocols that enable efficient recovery of epithelial cells. Here, we improve the canonical pipeline for single-cell analysis, optimizing the harvesting of nuclei from hard-to-dissociate tissues. We provide an in-depth mapping of the murine pancreas through paired RNA/ATAC multiomic single-nucleus sequencing, documenting the transcriptomic and chromatin accessibility profiles of every cell in the parenchyma. This enables a superior examination of the exocrine fraction of the pancreas. We described endocrine-exocrine interaction in silico and validated pro-proliferative signals in vitro. At the same time, we assessed pancreatic cellular heterogeneity in a holistic manner. We showed that pancreatic acinar cells can acquire multiple phenotypic states that lead to the functional diversification of pancreatic acini. In particular, we identified ultra-specialized ZG16HIGH cells in vivo and associated them with superior protein synthesis by tracking the incorporation of a synthetic amino acid. Leveraging the pairing of gene expression and chromatin opening, we studied the epigenetic elements that dictate acinar cell specialization and how they are affected by tissue repair after inflammation. We found that a subset of acinar cells shows enhanced sensitivity to inflammatory cues that determine extensive and persistent chromatin opening. The dataset is an open-source framework to interrogate the molecular histology of the murine pancreas.

Combined hepatocellular-cholangiocarcinoma (cHCC-CCA) is a rare primary liver cancer characterized by features of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA). As cHCC-CCA prognosis often falls between HCC and CCA or resembles CCA, we compared the outcomes of durvalumab plus gemcitabine-cisplatin (GCD) in patients with cHCC-CCA and CCA in a real-world setting. This retrospective multicenter study included patients with histologically confirmed unresectable or metastatic CCA or cHCC-CCA who underwent first-line GCD at four centers in Taiwan between 2021 and 2025. Outcomes included overall survival (OS), progression-free survival (PFS), tumor response, and safety. Propensity score matching (PSM) was performed to balance baseline characteristics. Among the 175 patients, 154 had CCA and 21 had cHCC-CCA. Before matching, patients with cHCC-CCA more frequently had elevated α-fetoprotein levels and poor biliary drainage; however, these imbalances were resolved after PSM. Before PSM, patients with CCA and cHCC-CCA had median OS of 13.3 months [95% confidence interval (CI) 9.5-17.1] and 13.8 months (95% CI 2.1-25.6, P = 0.60) and median PFS of 5.1 months (95% CI 4.7-5.5) and 5.3 months (95% CI 3.1-7.5, P = 0.40), respectively. After PSM, both OS and PFS rates were similar between the groups. The objective response rate was higher in the cHCC-CCA group (38.1% versus 23.4% before PSM, P = 0.15; 38.1% versus 23.8% after PSM, P = 0.24), whereas the disease control rates were identical (66.7% after PSM). The most common toxicities were hematologic events, with both groups demonstrating similar rates of anemia, thrombocytopenia, leukopenia, and neutropenia. This study demonstrates that patients with cHCC-CCA derive comparable clinical benefit from GCD as those with CCA in terms of efficacy and safety. These findings support the incorporation of immunochemotherapy into the treatment framework for cHCC-CCA and provide a rationale for its use in clinical practice.

Treatment options for advanced biliary tract cancers (BTCs) progressing after first-line chemoimmunotherapy remain limited. Whether platinum-based chemotherapy reintroduction can restore disease control is unknown. This study evaluates cisplatin-gemcitabine-durvalumab (CGD) reintroduction in patients with advanced BTC progressing during durvalumab maintenance. A multinational cohort of patients with BTC received first-line CGD followed by durvalumab maintenance. Those progressing during maintenance and treated with CGD reintroduction or FOLFOX/XELOX were analyzed. Primary endpoints were overall survival (OS) and progression-free survival (PFS) in the CGD reintroduction group; secondary endpoints included survival outcomes from maintenance and second-line initiation, overall response rate, and disease control rate. Among 1258 patients treated with first-line CGD, 475 received durvalumab maintenance. After disease progression during maintenance (n = 309, 65.1%), 31 patients (6.5%) underwent CGD reintroduction, while 114 (24%) received FOLFOX/XELOX. Baseline characteristics were comparable between groups, except for bilirubin levels. After a median follow-up of 22.6 months [95% confidence interval (CI) 20.0-25.1], median OS was not reached in the reintroduction cohort, while median PFS was 13.3 months (95% CI 9.7-15.8) from first-line initiation. From durvalumab maintenance start, median OS remained not reached and median PFS was 7.1 months (95% CI 4.8-11.3). Following reintroduction, median OS and PFS were 10.5 months (95% CI 8.0-10.5) and 9.0 months (95% CI 5.5-10.2). Compared with FOLFOX/XELOX, CGD reintroduction was associated with significantly improved OS [hazard ratio (HR) 0.47; P = 0.006] and PFS (HR 0.44; P < 0.0001) from first-line therapy start, as well as improved PFS from second-line treatment start (HR 0.60; P = 0.027). These findings were confirmed after multivariable and inverse probability of treatment weighting adjustment. Response outcomes were comparable between groups. This is the first study evaluating CGD reintroduction in patients with BTC progressing during durvalumab maintenance. Although the retrospective design does not allow definitive conclusions, our findings suggest potential benefit in selected patients.

Primary Sclerosing Cholangitis (PSC) is a chronic cholestatic liver condition that is closely associated with inflammatory bowel disease (IBD). PSC is associated with substantial long-term hepatobiliary morbidity, including an increased risk of cholangiocarcinoma. Reported prevalence and incidence estimates of PSC vary widely across different populations and study designs. This meta-analysis aims to provide an updated estimate of the prevalence of PSC in the general and IBD populations, and PSC incidence in the general population. A systematic search of MEDLINE (Ovid) and Embase from inception to October 20, 2025, for population- or cohort-based studies reporting PSC prevalence or incidence was performed. Seventy-four studies met the inclusion criteria. Random-effects meta-analyses were performed to pool prevalence and incidence estimates. Subgroup analyses, meta-regression, and sensitivity analyses were conducted to assess robustness and sources of heterogeneity. Seventeen studies including 196,635,709 individuals showed a pooled overall PSC prevalence of 8.06 per 100,000 (95% CI 4.75-13.36) in the general population. Among 516,548 patients with IBD in thirty-four studies, pooled PSC prevalence was 15.2 per 1,000 (95% CI 10.7-21.7). PSC prevalence was higher in ulcerative colitis (20.6 per 1,000) than in Crohn's disease (10.2 per 1,000). Using Crohn's disease as the reference group, ulcerative colitis was associated with higher odds of PSC (OR 1.787, 95% CI 1.066-2.996, p = 0.028). Thirteen studies reported a pooled incidence of PSC in the general population of 0.892 per 100,000 person-years (95% CI 0.705-1.130). This meta-analysis provides an updated synthesis of PSC epidemiology, extending prior work by incorporating detailed subgroup analyses across IBD phenotype, PSC subtype, geographic region, socioeconomic context, diagnostic modality, case-ascertainment method, and time period, which are dimensions not comprehensively addressed in previous meta-analyses. PSC remains a rare disease in the general population but is substantially more prevalent among individuals with IBD. Marked geographic and methodological heterogeneity, with notable underrepresentation of data from lower-SDI regions and the African Region, underscores the need for standardized case definitions and improved epidemiological surveillance across diverse settings. Given the markedly increased risk of cholangiocarcinoma in PSC, these findings highlight the clinical importance of recognizing PSC in higher-risk IBD populations and provide an epidemiological foundation for future studies evaluating risk stratification, early detection, and surveillance.

In the TOPAZ-1 trial's primary analysis, durvalumab plus gemcitabine and cisplatin (GemCis) showed statistically significant improved overall survival (OS) vs placebo plus GemCis with comparable safety between treatment groups in participants with advanced biliary tract cancer (aBTC). Durvalumab plus GemCis was recently established as the first-line standard of care among patients with aBTC. To evaluate 4-year OS and safety of durvalumab plus GemCis in participants with aBTC. This post hoc analysis of the global, double-blind, placebo-controlled, phase 3 TOPAZ-1 randomized clinical trial included participants 18 years and older with histologically confirmed unresectable, locally advanced, or metastatic biliary tract adenocarcinoma. In the TOPAZ-1 trial, patients were enrolled from April 2019 to December 2020 at 105 sites in 17 countries. Data cutoff was February 28, 2025. Participants received intravenous durvalumab, 1500 mg, or placebo plus gemcitabine, 1000 mg/m2, and cisplatin, 25 mg/m2, on days 1 and 8 every 3 weeks for up to 8 cycles, followed by durvalumab or placebo monotherapy every 4 weeks. OS, duration of treatment exposure, serious adverse events, and adverse events resulting in discontinuation were assessed approximately 48 months after the last participant was randomized. Overall, 685 participants were randomized, with 341 receiving durvalumab plus GemCis (median [range] age, 64 [20-84] years; 172 [50.4%] female) and 344 receiving placebo plus GemCis (median [range] age, 64 [31-85] years; 168 [48.8%] female). Median (range) follow-up in censored participants was 56.9 (1.7-67.2) months for participants who received durvalumab plus GemCis and 50.7 (0.9-62.6) months for participants who received placebo plus GemCis. Median OS was 13.0 (95% CI, 11.6-14.1) months for durvalumab plus GemCis and 11.4 (95% CI, 10.1-12.5) months for placebo plus GemCis (hazard ratio, 0.75; 95% CI, 0.64-0.88); 48-month OS rate was 11.8% vs 4.3%, respectively. The rate of serious adverse events possibly related to treatment was similar between arms (52 of 338 participants [15.4%] in the durvalumab plus GemCis arm vs 59 of 342 participants [17.3%] in the placebo plus GemCis arm). In the durvalumab plus GemCis and placebo plus GemCis arms, 21 of 338 participants (6.2%) and 18 of 342 participants (5.3%), respectively, experienced adverse events leading to study drug discontinuation. In this post hoc analysis of the phase 3 TOPAZ-1 randomized clinical trial, durvalumab plus GemCis demonstrated long-term survival benefit and a clinically manageable safety profile, supporting its use as a first-line treatment for aBTC. ClinicalTrials.gov Identifier: NCT03875235.

Patients with advanced biliary tract cancers (aBTC) are in urgent need of additional/new treatment options. We aimed to assess the efficacy and safety of ivonescimab plus chemotherapy in patients with aBTC. In this multicenter, open-label, phase II study, 30 patients with treatment-naive unresectable locally advanced or metastatic BTC received ivonescimab (20 mg/kg or 30 mg/kg) combined with gemcitabine (1000 mg/m2) and cisplatin (25 mg/m2) every 3 weeks for up to eight cycles, followed by ivonescimab maintenance. The primary endpoint was the investigator-assessed objective response rate (ORR) and safety. Pretreatment tumor specimens available from the trial were subjected to a post hoc exploratory proteomic analysis. The correlation between MAP2K7 levels and ivonescimab efficacy was assessed by BTC tumor cell-T cell co-culture and BTC organoids-T cell. At data cutoff, 1 patient achieved complete response and 19 patients achieved partial response yielding an ORR of 66.7% (95% Confidence Interval [CI]: 47.2-82.7). The disease control rate was 100%. The median progression-free survival (mPFS) was 8.5 months (95% CI: 7.6-10.5) and the median overall survival (mOS) was 16.8 months (95% CI: 11.1-22.5). Treatment-related adverse events occurred in 100.0% of patients with the most common being anemia (25, 83.3%), neutrophil count decreased (23, 76.7%), white blood cell count decreased (22, 73.3%), and platelet count decreased (22, 73.3%). No treatment-related deaths occurred. Additionally, exploratory proteomic and functional analyses identified MAP2K7 as a resistance-associated biomarker. MAP2K7 was upregulated in non-responders, and MAP2K7 suppression enhanced ivonescimab-mediated antitumor activity in immune co-culture models. Ivonescimab plus chemotherapy showed potential anti-tumor activity and tolerable safety as first-line treatment of aBTC patients. Exploratory analyses suggest that MAP2K7 may serve as a candidate biomarker of resistance and a potential therapeutic target for optimizing ivonescimab-based therapy of aBTC patients. Currently, the standard of care for first-line therapy in patients with aBTC is the addition of immune checkpoint inhibitors to chemotherapy based on the TOPAZ-1 and KEYNOTE-966 trials. However, this new regimen only improved OS by less than 2 months. The ORR of 66.7%, DCR of 100% and the median overall survival of 16.8 months were observed with ivonescimab plus chemotherapy. This study provides evidence supporting the potential role of ivonescimab plus chemotherapy as a first-line therapy for patients with treatment-naive unresectable locally advanced or metastatic BTC. NCT05214482 and NCT06048289. NCT05214482 and NCT06048289.

TOPAZ-1 (NCT03875235) demonstrated improved overall survival (OS) with durvalumab plus gemcitabine and cisplatin vs. placebo plus gemcitabine and cisplatin with manageable safety in advanced biliary tract cancer (aBTC). The phase IIIb TOURMALINE study (NCT05771480) is evaluating the safety and efficacy of first-line durvalumab plus seven gemcitabine-based chemotherapy regimens in a close-to-real-world aBTC population, including participants with poor prognosis. Participants received 1500 mg IV durvalumab (first infusion: 60 minutes; subsequent infusions: 30 minutes) plus investigator's choice of one of seven gemcitabine-based chemotherapy regimens. The primary endpoint was incidence of Grade 3/4 adverse events (AEs) possibly related to treatment (PRAEs) within 6 months of first durvalumab dose. Secondary endpoints included progression-free survival (PFS), OS, objective response rate (ORR), and safety. As of 17 September 2025, 142 participants (median age 68.0 years) had follow-up ≥6 months and received ≥1 treatment dose; most had metastatic disease (72.5%). Participants received a median of 9 cycles of durvalumab; median (IQR) follow-up was 11.63 (6.87-17.25) months. Overall, 50.7% (95% confidence interval [CI] 42.19-59.19) of participants experienced Grade 3/4 PRAEs within 6 months of durvalumab initiation, with no serious AEs of special interest with an outcome of death. Median PFS was 7.39 months (95% CI 6.74-9.07); median OS was 13.50 months (95% CI 11.24-20.53); ORR was 33.1% (95% CI 25.44-41.48). Safety profiles of durvalumab plus seven gemcitabine-based chemotherapy regimens were manageable, and shorter durvalumab infusion duration (30 minutes vs. 60 minutes) did not increase infusion-related reactions. Efficacy data are indicative of potential clinical activity across treatment arms. These findings demonstrate the feasibility of combining durvalumab with alternative gemcitabine-based regimens in a broad aBTC population. The choice of gemcitabine-based chemotherapy backbones for the treatment of advanced biliary tract cancer (aBTC) varies globally and their use in conjunction with immunotherapy has not been fully investigated. The TOURMALINE phase IIIb study demonstrated manageable safety and efficacy data showed potential clinical activity in a close-to-real-world population of participants with aBTC receiving durvalumab plus one of seven gemcitabine-based chemotherapy regimens. These findings point towards the feasibility of combining durvalumab with different gemcitabine-based chemotherapy regimens, even in participants with less favourable disease characteristics. NCT05771480 TRIAL REGISTRATION: ClinicalTrials.gov: NCT05771480; https://clinicaltrials.gov/study/NCT05771480.

FGFR2 fusion is a common alteration in malignancies, including intrahepatic cholangiocarcinoma (iCCA). While FGFR2 fusion has oncogenic properties, iCCA patients harboring this alteration demonstrate favorable prognosis, remaining a paradox. Here, we delineated the transcriptomic landscape of FGFR2 fusion-positive iCCA. We observed transcriptional features related to PI3K-AKT signaling in tumor cells and reduced neutrophil infiltration, particularly of the PD-L1+ neutrophil subtype. Mechanistically, FGFR2 fusion was associated with reduced H3K27ac enrichment at the CXCL3 promoter and decreased CXCL3 expression in tumor cells, which may contribute to impaired neutrophil recruitment to tumor tissues. In preclinical models, pharmacological FGFR inhibition increased CXCL3 levels and neutrophil infiltration. Combining neutrophil blockade with clinically available FGFR inhibitors enhanced antitumor activity. In conclusion, this study provides mechanistic insights into the paradox between the oncogenic properties of FGFR2 fusion and favorable clinical outcomes in FGFR2 fusion-positive iCCA, and suggests a potential strategy to optimize FGFR2-targeted therapies.

Identifying diagnostic and prognostic biomarkers and therapeutic targets for hepatocellular carcinoma (HCC) is essential to improve risk stratification, guide individualized treatment, and enhance therapeutic efficacy.The expression of SAMM50 (Sorting and Assembly Machinery Component 50) was initially analyzed in publicly accessible curated genomic and proteomic databases, such as the Cancer Cell Line Encyclopedia, the Human Protein Atlas, and other HCC-specific repositories. This analysis revealed differential expression patterns between HCC and non-neoplastic liver tissue. Subsequently, clinicopathological data and tissue specimens were collected from 200 HCC patients who underwent treatment at our institution. The protein and transcript levels of SAMM50 were experimentally measured in paired HCC and adjacent non-tumorous tissues using immunohistochemistry (IHC) and quantitative reverse transcription polymerase chain reaction (qRT-PCR). The association between SAMM50 expression and key clinicopathological features was further evaluated. Univariate and multivariate Cox proportional hazards analyses were performed to determine the independent prognostic value of SAMM50 expression in HCC. Based on these results, a reproducible and clinically applicable nomogram, supported by a forest plot, was constructed to facilitate prognostic prediction and support individualized therapeutic decision-making. Finally, in vitro and in vivo experiments were conducted to characterize the phenotypic alterations in HCC cells after SAMM50 knockdown, thereby confirming its involvement in critical oncogenic behaviors.This research demonstrated that the mRNA and protein levels of SAMM50 in HCC tissues were elevated compared to those in normal liver and adjacent tissues. Immunohistochemistry findings confirmed that SAMM50 protein levels were persistently higher in HCC tissues than in paired adjacent tissues. High expression of SAMM50 was correlated with unfavorable clinicopathological factors, encompassing pretreatment alpha-fetoprotein (AFP) levels, tumor size, T stage, American Joint Committee on Cancer (AJCC) stage, histological grade, and worse overall survival.Specifically, high expression of SAMM50 was linked to shorter overall survival (OS), progression-free survival (PFS), and disease-free survival (DFS). Moreover, univariate and multivariate Cox analyses were conducted to investigate the association between SAMM50 expression and clinicopathological features in HCC patients and to identify independent prognostic factors. The area under the receiver operating characteristic (ROC) curve (AUC) for SAMM50 was 0.863, suggesting its potential as a diagnostic marker for HCC, though further validation in independent cohorts is needed. Silencing of SAMM50 inhibited HCC cell proliferation, migration, and invasion, promoted apoptosis in vitro, and suppressed HCC growth in vivo.This research demonstrates that SAMM50 shows potential diagnostic value for HCC, though this observation requires further validation in larger, independent, and prospective cohorts. The results of this study not only contribute to the evaluation of baseline data and risk stratification in HCC but also offer novel approaches for the development of precise treatment strategies and targeted therapies.

The comparative outcomes of entecavir (ETV), tenofovir disoproxil fumarate (TDF), and tenofovir alafenamide (TAF) in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) after curative resection remain unclear. This study aimed to evaluate the survival benefits of these antiviral regimens in a large real-world cohort. We analysed 902 patients with BCLC stage 0 or A HBV-related HCC who underwent curative resection between September 2001 and August 2025. Patients received ETV (n = 474), TDF (n = 363), or TAF (n = 65) as first-line therapy within 3 months post-resection. Inverse probability of treatment weighting (IPTW) was implemented to balance baseline covariates. The primary and secondary endpoints were overall survival (OS) and recurrence-free survival (RFS). The IPTW-adjusted 5-year OS rates for ETV, TDF, and TAF were 90.7%, 95.2%, and 97.1%, respectively (global p = 0.079). The 5-year RFS rates were comparable for ETV, TDF, and TAF (57.4% vs. 61.0% vs. 69.4%; global p = 0.533). In multivariable Cox regression analysis, however, TDF was independently associated with a significantly lower risk of death compared with ETV (aHR 0.48; 95% CI, 0.26-0.89; p = 0.021). TAF showed higher survival rates compared to ETV, although the difference did not reach statistical significance (aHR 0.34; 95% CI, 0.04-2.55; p = 0.292). The choice of antiviral regimen was not associated with RFS after curative resection for HBV-related HCC, whereas TDF was independently associated with improved OS compared with ETV. Further large-scale studies are warranted to validate the long-term prognostic impact of TAF.

Risk stratification in hepatocellular carcinoma (HCC) is limited by the lack of robust biomarkers reflecting tumor biology. The antioxidant enzyme Paraoxonase-1 (PON1) shows prognostic potential, yet its role in tumor tissues and the feasibility of non-invasive assessment remain unclear. Senescence-related pathways and prognostic candidates were screened using transcriptomic data from TCGA-LIHC and GTEx. PON1 expression was validated in multicenter cohorts through qPCR, immunohistochemistry, and Western blotting. Functional assays in PON1 knockdown and overexpression models evaluated oxidative stress, glutathione balance, mitochondrial dysfunction, and senescence markers. We developed a radiomics model based on contrast-enhanced CT scans with super-resolution reconstruction to predict tumoral PON1 expression. This radiomics signature was then integrated with clinical variables to build a combined model, which was evaluated across training, validation, test, and external cohorts. PON1 was identified as a downregulated senescence-related prognostic gene. Low PON1 expression was associated with poorer overall and progression-free survival across independent clinical cohorts. PON1 depletion increased intracellular and mitochondrial ROS, lowered the GSH/GSSG ratio, impaired mitochondrial membrane potential, and induced senescence phenotypes, while its restoration mitigated these effects. SR-enhanced radiomics improved prediction of tumoral PON1 expression across all cohorts. Integration of radiomics signatures with clinical variables further improved discrimination, achieving the highest accuracy and net clinical benefit. PON1 downregulation contributes to oxidative stress-driven senescence and unfavorable clinical outcomes in HCC. SR-enhanced radiomics provides an accurate, non-invasive method for estimating tumoral PON1 expression, demonstrating potential value for radiogenomic profiling and preoperative risk stratification.

The efficacy of tyrosine kinase inhibitor (TKI)-based systemic therapy in advanced hepatocellular carcinoma (HCC) is often limited by drug resistance, the mechanisms of which remain incompletely understood. Here, we demonstrate that CD276, an immune checkpoint protein, promotes TKI resistance in HCC by reprogramming lipid metabolism. Upon TKI treatment, CD276 binds pSTAT3 and undergoes importin α/β-dependent nuclear translocation. In the nucleus, CD276 cooperates with pSTAT3 to promote CD36 transcription, thereby potentiating fatty acid uptake, lipid droplet accumulation, and mitochondrial fatty acid β-oxidation. This metabolic rewiring drives HCC proliferation and confers TKI resistance. Importantly, pharmacological inhibition of CD36 with sulfosuccinimidyl oleate sodium suppresses fatty acid uptake and tumor lipid metabolism, resensitizing resistant HCC cells to TKIs. Our findings reveal the CD276-pSTAT3-CD36 axis as a key regulator of lipid metabolic reprogramming in TKI resistance, providing a promising therapeutic target to overcome treatment resistance in HCC.

Laparoscopic liver resection (LLR) is challenging due to the complex and variable intrahepatic vascular anatomy, limited surgical field of view, and lack of tactile feedback, which collectively increase the risk of intraoperative injury. Accurate identification of anatomical structures is therefore essential for safe LLR. This study explores Vivim, a video segmentation model based on the Mamba architecture-a state-space model that integrates the long-range dependency modeling of transformers with the efficiency of recurrent networks. Evaluated on a multicenter dataset of 15,865 annotated frames from 45 videos, Vivim outperformed several image-based and video-based baselines, including U-Net, DeepLab v3 + , nnU-Net, FPN + EfficientNetV2-L, SegFormer, ConvLSTM-U-Net, ViViT and SegFormer + T-Mamba, in segmenting the Glissonean pedicle (GP) and hepatic vein (HV). By leveraging Mamba's selective scanning and linear complexity, Vivim achieved Dice scores of 0.71 and 0.66 in single- and multi-target segmentation tasks, respectively, while maintaining real-time inference at 25 fps. The model demonstrated strong generalization on a cross-center external test set and robustness to typical surgical challenges. Clinically validated by 13 surgeons, Vivim improved recognition speed and accuracy, aiding intraoperative decision-making. Despite difficulties in differentiating similar vessels, this Mamba-based framework represents a promising step toward AI-assisted surgical navigation, bridging laboratory precision and clinical reliability.

As trypsin activation occurs early in the pathogenesis of pancreatitis, evaluating serum trypsin (TRY) may allow immediate prediction of post-ERCP pancreatitis (PEP). This study aims to evaluate the diagnostic performance of TRY measurement for predicting PEP. This multicenter prospective observational study was conducted at 12 tertiary centers in Japan. Serum TRY, amylase (AMY), pancreatic-type AMY (P-AMY), and lipase (LIP) were measured before ERCP, immediately post-procedure (0 h), and 2 h post-procedure. Predictive performances of pancreatic enzymes for PEP were evaluated using receiver operating characteristic (ROC) analysis, Youden's index, and DeLong's test. PEP was identified in 48 of the 463 patients analyzed (10.3%). The optimal 0-h TRY cut-off was 2043 ng/mL, yielding an area under the ROC curve (AUC) of 0.83 (68.7% sensitivity, 84.6% specificity), significantly higher than those of 0-h AMY (AUC 0.74, p = 0.002), 0-h P-AMY (0.78, p = 0.007), or 0-h LIP (0.80, p = 0.048). At 2-h, TRY showed an AUC of 0.89, superior to 2-h AMY (0.84) and comparable to 2-h P-AMY (0.89) and 2-h LIP (0.90). When cutoffs were set as 2 × , 2.5 × and 3 × of the upper limit of normal, TRY consistently showed higher sensitivity than AMY or P-AMY. TRY showed the earliest and steepest post-procedural increase among all enzymes in patients with PEP. TRY measured immediately after ERCP provides significantly better early prediction of PEP than conventional enzymes. Rapid measurement of TRY may enable immediate risk stratification, potentially improving clinical outcomes.

To investigate whether cyst shrinkage in intraductal papillary mucinous neoplasm (IPMN) is paradoxically associated with a higher risk of concomitant pancreatic ductal adenocarcinoma (PDAC). This retrospective cohort included 1103 patients with pathologically or cytologically confirmed branch-duct or mixed-type IPMN followed for ≥ 1 year. Patients were classified by annual cyst size change rate per the 2024 Kyoto guideline thresholds: shrinkage (< 0 mm/year), stable (0 to < 2.5 mm/year), and growth (≥ 2.5 mm/year). The primary outcome was concomitant PDAC, with IPMN-derived carcinoma as a competing event. Multivariable Fine-Gray regression compared shrinkage with a pre-specified merged non-shrinkage reference; cause-specific Cox regression served as sensitivity analysis. Of 1103 patients (541 men, 562 women; mean age, 67.3 ± 9.8 years), 282 (25.6%) showed shrinkage, 728 (66.0%) remained stable, and 93 (8.4%) showed growth. Concomitant PDAC was most frequent in shrinkage (6/282, 2.1%), followed by stable (8/728, 1.1%), with no events in growth (0/93). IPMN-derived carcinoma was most frequent in growth (9/93, 9.7%). Adjusted for age, sex, initial cyst size, and main pancreatic duct diameter, shrinkage was independently associated with concomitant PDAC (subdistribution hazard ratio 4.46; 95% CI 1.62-12.30; p = 0.004); cause-specific Cox regression yielded a concordant estimate (hazard ratio 4.54; 95% CI 1.52-13.50; p = 0.007). In this single-center retrospective cohort, cyst shrinkage was associated with an increased incidence of concomitant PDAC, contrary to the prevailing assumption that shrinkage reflects a benign course. Given the modest number of concomitant PDAC events (n = 14), these findings are hypothesis-generating and exploratory; external validation in multicenter prospective cohorts is required before clinical surveillance practice is modified. Question Cyst shrinkage during intraductal papillary mucinous neoplasm surveillance is considered reassuring, but its association with concomitant pancreatic cancer risk remains unclear. Findings Shrinkage was paradoxically associated with higher concomitant pancreatic cancer risk (subdistribution hazard ratio 4.46); the growth group's 0% rate reflected competing risks from IPMN-derived carcinoma. Clinical relevance Radiologists should not reduce surveillance intensity for shrinking intraductal papillary mucinous neoplasm cysts, as these patients carry an elevated risk of concomitant pancreatic cancer developing independently of the monitored cyst.

Early detection of focal liver lesions is critical for patient outcomes, but current imaging techniques have limitations. This study developed MULLET, an AI system using contrast-enhanced CT, and assessed its performance in assisting radiologists with FLL detection and characterization. A retrospective multicenter, multi-reader, multi-case trial was conducted, in which 10 radiologists independently interpreted 375 patients' clinical images with and without MULLET assistance. This trial was registered on ClinicalTrials.gov (ID: NCT06068413) with the registration name "A Retrospective, Multicenter, Multiple-viewer-multiple-case (MRMC) Clinical Trial to Evaluate the Safety and Efficacy of CT Image-assisted Detection Software for Focal Liver Lesions" in April 2023. Diagnostic performance was compared using area under the receiver operating characteristic curve (AUC) analysis. Without MULLET, the average AUC was 0.8188 (sensitivity: 74.45%, specificity: 87.77%). With MULLET, the average AUC significantly improved to 0.9268 (p < 0.0001), sensitivity increased to 89.95% (p = 0.0003), and specificity rose to 93.57% (p = 0.0063). MULLET also showed improved performance across different FLL sizes and subtypes. Moreover, the average reading time for radiologists was reduced by 21.8% (p < 0.0001). In conclusion, MULLET demonstrated promising performance in significantly improving radiologists' sensitivity, accuracy, and efficiency in FLL detection and diagnosis.

Locoregional therapy (LRT) is frequently used as bridging therapy to transplantation/resection or palliative treatment for hepatocellular carcinoma (HCC). Multiple prediction models have been developed for prognosis and treatment response among patients undergoing LRTs. We aimed to systematically review the methodological quality and performance of clinical risk scores predicting outcomes in patients with HCC treated with LRT. EMBASE and PubMed were searched from inception to 18 March 2026. Our main outcome was the concordance statistic, or area under the receiver operating characteristic curve (AUROC), to predict survival and other tumour-related and liver-related outcomes. 130 studies met the inclusion criteria, resulting in 179 individual scoring systems. The total population was 70 061 patients, 21% female (4-51%), most with Barcelona Clinic Liver Cancer Stage B (39%). Risk scores commonly incorporated tumour parameters, liver function tests, cirrhosis staging and comorbidities. AUROC values ranged from 0.56 to 0.94. 16 studies (12.3%) had low risk of bias, while most had high risk of bias. Of these, the Y-scoring system, Cheng et al nomogram and Li et al nomogram showed the highest discrimination for overall survival in palliative LRT (AUROC >0.87). Separate pooled analyses showed that the Hepatoma Arterial-embolisation Prognostic (HAP) and modified Hepatoma Arterial-embolisation Prognostic II (mHAP-II) scores had the highest performance (pooled AUROC >0.72), while the Six-and-Twelve (0.68) and Albumin-Bilirubin (ALBI) scores (0.60) demonstrated lower performance. Several outcome scoring systems are promising for specific LRTs and populations. However, most models have high risk of bias, underscoring the need for further development and validation.

Post-treatment surveillance after locally treated colorectal liver metastases involves CEA measurement and contrast-enhanced CT (CECT) scans. However, reactive tissue around ablated lesions can appear similar to viable tumor tissue on CECT, complicating the detection of local tumor progression (LTP). Therefore, a new imaging protocol was initiated, which incorporates [18F]FDG-PET/CT to complement CECT. This study evaluated the diagnostic accuracy of [18F]FDG-PET/CECT for early detection of disease progression after resection or thermal ablation, compared to CECT alone. The study analyzed a prospective cohort of patients undergoing thermal ablation or resection, who received CECT and [18F]FDG-PET/CT scans 3-5 months post-treatment. Disease progression was confirmed by histology, consensus during a multidisciplinary team meeting or after extended follow-up. Diagnostic accuracy of [18F]FDG-PET/CECT was compared to standard practice (CEA serum levels and CECT). The final analysis included 64 patients with 154 lesions. 37 patients underwent ablation, 14 patients underwent resection, and 13 patients underwent combination therapy, resulting in 93 (60%) lesions being treated with thermal ablation and 61 (40%) lesions with resection. Disease progression was found in 66% of patients, with LTP detected in 25% of treated lesions. Compared to CECT, [18F]FDG-PET/CECT had a higher diagnostic accuracy for detecting local progression after thermal ablation (AUC 0.97 vs. 0.73, p = 0.001) and hepatic resection (AUC 0.96 vs. 0.69, p = 0.03), and for detection of extrahepatic disease (AUC 0.91 vs. 0.79, p = 0.03). CEA serum levels had low diagnostic accuracy for detecting disease progression (AUC 0.60). [18F]FDG-PET/CECT improves diagnostic accuracy in the early follow-up of patients with locally treated colorectal liver metastases. Question: Interpreting CECT scans after local treatment of colorectal liver metastases can be challenging because reactive tissue around treated lesions can appear similar to viable tumor tissue. [18F]FDG-PET/CECT improves the diagnostic accuracy of detecting local tumor progression after thermal ablation and resection of colorectal liver metastases. [18F]FDG-PET/CECT should be considered during follow-up after local treatment of colorectal liver metastases to identify and treat local tumor progression earlier.

The role of primary tumor surgery in patients with de novo metastatic soft tissue sarcoma (STS) is controversial, with a lack of objective selection criteria. This study aimed to identify patients who may benefit from surgery and to evaluate the prognostic value of integrated 18F-FDG PET/CT metabolic parameters and clinicopathological factors. Forty patients with de novo metastatic STS undergoing pretreatment ¹⁸F-FDG PET/CT were analyzed. Clinical data and metabolic parameters (maximum standardized uptake value [SUVmax], total metabolic tumor volume, and total lesion glycolysis [TLG]) were collected. Prognostic factors for overall survival (OS) were assessed using Cox regression. Heterogeneity was examined through pre-specified subgroup analyses and formal interaction tests. Multivariate analysis identified age > 50 years, lactate dehydrogenase (LDH) > 200 U/L, and SUVmax > 10.0 as independently associated with worse OS (all p < 0.05). Significant interactions were observed between surgical treatment and both the Ki67 index and TLG (P for interaction = 0.006 and 0.042, respectively). Surgery was associated with significantly improved OS in patients with low Ki67 (HR = 0.24, 95% CI: 0.06-0.89; p = 0.032) or low TLG (HR = 0.21, 95% CI: 0.04-0.97; p = 0.046), with no significant benefit observed in corresponding high-expression subgroups (all p > 0.05). The survival benefit of primary tumor surgery in de novo metastatic STS is restricted to patients with less aggressive tumor dbiology, characterized by low proliferative activity (Ki67 ≤ 35%) or low TLG. Integrated assessment of Ki67 and PET/CT parameters, particularly TLG, provides a practical framework for personalized surgical decision-making. Question Does primary tumor surgery provide a survival benefit for patients with de novo metastatic STS based on PET/CT and clinicopathological markers? Findings Surgery was associated with improved survival only in patients with low Ki67 or low TLG, with significant interaction effects (p = 0.006 and 0.042, respectively). Clinical relevance Surgical selection in metastatic STS should be guided by tumor biology. Integrating Ki67 and TLG provides a practical framework to identify patients most likely to derive a survival benefit from primary tumor resection.

Comprehensive genomic profiling (CGP) is widely used to identify actionable alterations and guide precision oncology, yet only a minority of tested patients receive genome-matched therapies, underscoring a gap between genomic findings and clinical benefit. We hypothesized that this gap may partly reflect variation in the reliability and interpretability of genomic information generated from specimens of different quality and by different assay modalities. To examine this possibility, we performed a retrospective multicenter analysis of 2002 CGP tests conducted between 2019 and 2025 across 13 institutions in Japan. Detection of short variants, copy number alterations (CNAs), structural variants, and genomic signatures, including microsatellite instability, tumor mutational burden, and homologous recombination deficiency signature, was compared among FoundationOne CDx specimens classified as pass (F1-pass) or qualified (F1-qual) and liquid-based CGP (liq-CGP). Short variant detection remained largely preserved in F1-qual specimens, whereas CNA and genomic signature detection were substantially reduced. In pancreatic adenocarcinoma, KRAS variants were detected in 93% of F1-pass, 88% of F1-qual, and 57% of liq-CGP cases. These differences affected the proportion of patients offered genome-matched therapies. Machine learning models predicted QC status with area-under-the-curve values of 0.72-0.78. Our findings support QC-aware CGP selection in routine precision oncology.

The prevalence of type 2 diabetes mellitus (T2DM) is rising rapidly in China and is linked to increased cancer risk, but causality remains unclear due to biases. We examined the causal effect of T2DM on cancer risk using bias-minimizing methods. We conducted a matched cohort study within China Kadoorie Biobank (median follow-up 49 months in men and 53 months in women). To minimize bias, we included only new-onset T2DM, applied sequential longitudinal matching, used pre-diagnosis BMI, and accounted for detection time bias. Stratified Cox models estimated sex-specific time-split hazard ratios (tsHRs). Results were triangulated with two-sample Mendelian Randomization (MR) in East Asians. After 1:3 matching, 8657 men and 13,680 women with T2DM were matched to unexposed individuals. T2DM was associated with increased risk of total cancers in men (tsHR 1.57, 95% CI 1.38-1.78) and women (tsHR 1.29, 95% CI 1.14-1.46). Site-specifically, T2DM was associated with liver cancer (men: 68 cases, tsHR 2.12, 95% CI 1.42-3.15; women: 43 cases, tsHR 2.39, 95% CI 1.42-4.04) and pancreatic cancer (men: 36 cases, tsHR 2.57, 95% CI 1.35-4.92; women: 33 cases, tsHR 3.95, 95% CI 1.92-8.13). No significant associations were observed for colorectal, lung, stomach, or breast cancers. In multivariable MR, genetic liability to T2DM was associated with pancreatic cancer after adjusting for BMI (OR 1.07, 95% CI 1.01-1.14, p = 0.03). In this large Chinese population, we found evidence for causal associations between T2DM and pancreatic cancer only, whereas evidence for other cancers was weak or discordant.

Reports on socioeconomic inequalities in cancer mortality are limited in East Asia. We investigated educational inequalities in cancer mortality in Japan, serving as an advanced example of a nationwide census-based surveillance. We developed a Japanese census-linked mortality dataset, using our unique linkage method. The dataset encompassed approximately 0.48 million cancer deaths linked to individual-level census data of 80 million Japanese aged 25-84 years in October 2020. We calculated age-standardised all-cancer and 23 site-specific cancer mortality rates (ASMRs) by educational level. Educational inequalities were quantified using the Relative Index of Inequality (RII) and Slope Index of Inequality (SII) by educational level (high, middle and low). Site-specific cancer contribution to absolute educational inequalities in all cancers was evaluated using the proportions of SII (%: SIICancer-site/SIIAll-cancer×100). All cancers' RIIs were 1.58 (95% CI 1.56 to 1.60) and 1.43 (1.40 to 1.46) in men and women, respectively. Among men, the rectum, stomach, liver and lung were the leading sites based on site-specific cancer RIIs, while the larynx, cervix uteri, liver and lung were the leading sites among women. In absolute terms, lung cancer contributed the most to educational inequalities, followed by stomach, colorectal and liver cancers. No educational inequality was found for pancreatic cancer. Breast cancer showed higher ASMRs among women with low education levels than among those with high education levels, which differed from the previous inequality pattern. Equity-focused strategies to enhance primary and secondary prevention are essential and should be supported by comprehensive nationwide monitoring.

Comprehensive genomic profiling (CGP) tests can identify putative biomarkers for immune checkpoint inhibitors (ICIs), including tumor mutational burden (TMB); however, their clinical utility remains uncertain. In this study, the efficacy and safety of nivolumab treatment based on the CGP results were investigated using a subcohort from the BELIEVE trial (NCCH1901; jRCTs031190104). This trial aims to improve drug accessibility for patients with advanced solid tumors harboring actionable genetic variants through off-label drug administration. Patients for whom off-label nivolumab treatment was proposed based on CGP results were eligible. Nivolumab was administered until disease progression or unacceptable toxicity. The primary endpoint was the best objective response rate (ORR) in patients with measurable disease. Secondary endpoints included progression-free survival (PFS), overall survival (OS), disease control rate (DCR), and safety. Among 60 enrolled patients, 56 patients who received study treatment were included in the analysis. TMB-High was the most frequent biomarker prompting nivolumab treatment (47/56, 83.9%), followed by CD274 (PD-L1) amplification (6/56, 10.7%) and CDK12 inactivating variants (3/56, 5.4%). Among 50 patients with measurable disease, the ORR was 16.0% (95% confidence interval [CI], 7.2-29.1) and the DCR was 40.0% (95% CI, 26.4-54.8). Median PFS and OS were 2.7 months (95% CI, 2.2-4.3) and 7.2 months (95% CI, 4.4-9.0), respectively. Although a subset of patients achieved durable responses, nivolumab treatment guided by CGP results demonstrated limited efficacy. The predictive value of TMB-High may differ across tumor types, and integration with other clinicogenomic factors is warranted to improve its predictive accuracy.

Multiple primary carcinomas (MPC) represent a clinically significant yet underexplored phenomenon, where patients develop more than one distinct primary malignancy. While prior studies have examined MPC within specific cancer types, comprehensive real-world patterns of primary malignancies and their subsequent primary malignancies remain limited. Moreover, the survival outcomes associated with these MPC patterns, particularly in relation to demographic and clinical characteristics, are not well characterized. This study aimed to establish the patterns and combinations of MPC across a wide range of cancers and to assess whether the mortality status of patients with MPCs varies according to their demographic characteristics and disease status. We conducted a retrospective analysis of 1560 patients with MPC in Shanghai, China, from 2002 to 2015. Data were extracted from the Shanghai Cancer Registry, with follow-up until December 2017. Cause of death was ascertained through linkage with the Shanghai Vital Registration System. The distribution of the frequency and proportion of primary carcinoma (PC) combinations were depicted, and a life table was used to calculate the 1- to 5-year survival rates. Cox regression analysis was performed to analyze the survival risk factors of the first and second PCs. Among the 1560 patients (809/1560, 51.86% male and 751/1560, 48.14% female), the most frequent first PCs were colorectal, breast, and stomach cancers, while the most frequent second PCs were lung, colorectal, and stomach cancers. The most common combinations included colorectal and lung, colorectal and stomach, and colorectal and prostate. Survival rates were lowest for first PCs of skin (5 years=46.95%) and lung (5 years=41.54%) cancers, and for second PCs of pancreatic (5 year=9.13%) and liver (5 years=14.19%) cancers. A latency period of 12 months between PC diagnoses was associated with significantly higher cancer-specific mortality for both the first primary cancer (hazard ratio [HR] 3.539, 95% CI 2.822-4.438; P<.001) and the second primary cancer (HR 1.369, 95% CI 1.103-1.699; P=.004). Older age (>65 years) and advanced tumor stage (III+IV) were also significant independent risk factors for poor survival in both first PC (age: HR 2.049, 95% CI 1.689-2.485; P<.001; stage: HR 1.496, 95% CI 1.315-1.703; P<.001) and second PC (age: HR 1.575, 95% CI 1.242-1.996; P<.001; stage: HR 3.933, 95% CI 3.182-4.861; P<.001) analyses. This study provides a comprehensive, real-world map of MPC patterns and highlights important findings: high-risk cancer combinations and key factors associated with poorer survival, including a short interdiagnosis interval (12 months), advanced age, and advanced tumor stage. Comprehensive prevention and control strategies for MPC should be developed, and clinicians should be aware of the risks of MPC in vulnerable populations during the early diagnosis stage.

Image-based machine learning tools are powerful resources for analyzing medical images, with deep learning-based semantic segmentation commonly utilized to enable the spatial quantification of structures visible in images. However, dataset generation and training of segmentation algorithms requires advanced programming skills and intricate workflows, limiting their accessibility to scientists without prior coding expertise. Here we present the step-by-step instructions to carry out automatic segmentation of medical images guided by a graphical user interface using the CODAvision algorithm. This workflow simplifies the process of semantic segmentation of microanatomical structures by enabling users to train highly customizable deep learning models without extensive coding expertise. The protocol outlines best practices for creating robust training datasets, configuring model parameters and optimizing performance across diverse biomedical image modalities. CODAvision enhances the usability of the CODA algorithm by streamlining parameter configuration, model training and performance evaluation, automatically generating quantitative results and comprehensive reports. We show the use of CODA to serial histology by demonstrating robust performance across numerous medical image modalities and diverse biological questions. We provide sample results in data types, including histology, magnetic resonance imaging and computed tomography. We demonstrate the diverse use of this tool in applications, including quantification of metastatic burden in in vivo models and deconvolution of spot-based spatial transcriptomics datasets. This protocol is designed for researchers with interest in rapid design of highly customizable semantic segmentation algorithms and a basic understanding of programming and anatomy.

Numerous liver diseases are characterized by late diagnosis, rapid progression and high incidence, seriously threatening public health. Though widely used, traditional treatments such as drug therapy, resection and transplantation have substantial limitations. Therefore, developing novel preventive strategies and specialized therapies is crucial. As Chinese medicine continues to modernize, increasing evidence suggests that certain Chinese medicine ingredients can protect the liver. Astragaloside IV (AS‑IV) is a natural saponin extracted from the root of the traditional herb Astragalus membranaceous. It exhibits diverse pharmacological activities, including anti‑inflammatory, antioxidant, antiapoptotic and anticancer properties, and is recognized for treating neurological, cardiovascular and metabolic disorders, and cancer. These discoveries indicate its substantial promise for the treatment of liver diseases. Therapeutic trials revealed its hepatoprotective effects for the treatment of various liver diseases, such as non‑alcoholic fatty liver disease, liver fibrosis, hepatocellular carcinoma and liver injury induced by heavy metals, drugs, or alcohol and involve various signaling pathways such as nuclear factor erythroid 2‑related factor 2, toll‑like receptor 4, acetyl‑CoA carboxylase, protein kinase B, nuclear factor κB and adenosine monophosphate‑activated protein kinase. The present study presents a narrative review that comprehensively summarizes existing evidence regarding the therapeutic influence of AS‑IV on diverse liver disorders and deeply analyzes the molecular mechanisms underlying its action in liver disease. The objective is to comprehensively offer insights and references for relevant scientific research and clinical drug development to improve nutritional supplements for liver health.

The prognosis for advanced colorectal cancer (CRC) remains poor, and 5-fluorouracil (5-FU)-based chemotherapy is the primary treatment option. Understanding the mechanisms that limit the effectiveness of this therapy is therefore clinically important. However, the molecular drivers of acquired chemotherapy resistance in advanced CRC are not fully understood. Here we show that expression of PDZ-LIM domain-containing protein 4 (PDLIM4) is significantly upregulated following neoadjuvant chemotherapy in patients with advanced CRC, and that its expression level is closely associated with patient prognosis. PDLIM4 promotes acquired resistance to 5-FU by enhancing the stemness, anti-apoptotic capacity, and drug efflux of colorectal cancer stem cells. Mechanistically, PDLIM4 facilitates the accumulation of sphingolipids, particularly sphingomyelin, and regulates the PI3K/Akt signaling pathway. These findings identify PDLIM4 as a potential biomarker and therapeutic target for overcoming chemoresistance in advanced colorectal cancer, opening new avenues for future treatment strategies.

Neuron-tumor communication is emerging as a distinct layer of tumor-host interaction beyond conventional stromal, vascular, and immune regulation. Recent studies show that malignant cells can detect neuronal activity and convert neural signals into growth-promoting cellular responses. In the brain, glioma cells can become electrically integrated into neuronal circuits, where glutamatergic input drives depolarization, calcium influx, and downstream signaling. In extracranial tumors, neural influence appears more heterogeneous, involving spatially organized neurochemical niches, receptor-enriched cancer-nerve contacts ('pseudo-synapses'), autonomic pathways, and injury-associated neuroimmune remodeling. These findings raise important questions about how neural input regulates tumor cell state, metabolism, immune tone, and therapeutic adaptation. This review evaluates the evidence linking neural activity to cancer progression across anatomical contexts and outlines the experimental standards needed to distinguish structured neuron-tumor interfaces from broader neural effects within the tumor microenvironment.

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

WNK2 is a known tumor suppressor in a set of solid tumors, including glioblastoma multiforme and pancreatic ductal adenocarcinoma. WNK2 functions are largely unknown beyond the kinase domain (KD)-dependent inhibition of ERK1/2 signaling. Waldenström's Macroglobulinemia (WM) is an indolent, yet incurable, B cell lymphoma characterized by highly recurrent MYD88 (MYD88MUT) and CXCR4 (CXCR4MUT) mutations that trigger sustained NF-κB and ERK1/2 signaling. Although not expressed in healthy B cells, WNK2 is a top dysregulated gene in MYD88MUT WM. To study WNK2 regulation and signaling, we performed multi-omics analyses, including bulk and PacBio Iso-Seq RNA-Seq and methylome, in 264 untreated WM and functional studies in cell lines and primary WM cells. Aberrant expression of WNK2 emerged as a near universal feature of early-stage WM and a hallmark of plasma cell-like MYD88MUT WM. We identified novel isoforms that carried a shared aberrant splicing event, either contained or lacked the KD and were highly expressed by the tumor cells, unlike the canonical full-length isoforms. Functionally, WNK2/S-NK1, the most expressed of the KD-lacking isoforms in WM, triggered a pro-inflammatory cascade that activated ERK1/2 and NF-κB signaling. We observed a similar, cancer-specific upregulation of WNK2 in a set of solid tumors, including the highly aggressive cholangiocarcinoma. Our findings reveal an undocumented oncogenic function for WNK2 driven by novel, cancer-specific isoforms and provide a framework for its further investigation as a determinant of disease progression in MYD88MUT WM and a novel therapeutic target in hematological and solid tumor oncology.

Toll-like receptor (TLR) signaling is critical for innate immune system. However, whether it is directly modulated by microbiota-derived metabolites remains unclear. Here, we show that the short-chain fatty acids (SCFAs) propionate and butyrate suppress TLR signaling by directly binding p38α MAP kinase, promoting its interaction with TAB1, thereby activating p38α via autophosphorylation. Activated p38α then phosphorylates TRAF3 at serine 85, inhibiting K63-linked polyubiquitylation of TRAF3 and disrupting TBK1-IRF3 activation, leading to reduced macrophage activation and intestinal inflammation. In ulcerative colitis patients, fecal levels of propionate and butyrate positively correlate with p38α activity and TRAF3 S85 phosphorylation, but inversely correlate with TBK1 activation, and cytokine levels. Notably, oral administration of propionate in three patients with ulcerative colitis markedly improved intestinal inflammation and clinical symptoms. These findings reveal p38α as a direct sensor for microbiota-derived SCFAs that suppress TLR signaling through nonmetabolic functions of propionate and butyrate, providing the first clinical evidence that propionate supplementation represents a practical dietary strategy for ulcerative colitis management.

Epithelial-to-mesenchymal transition (EMT) is a dynamic process during which cells lose their epithelial characteristics and acquire mesenchymal traits. In cancer, EMT is closely associated with tumor initiation, progression, invasion, metastasis, and therapy resistance. Rather than being a binary state switch, EMT encompasses a spectrum of tumor states with distinct functional properties. However, the transcription factors (TFs) that govern transitions between these EMT states remain poorly defined. Here, using multi-omic approaches combining single-cell RNA-seq and single-cell ATAC-seq, we delineate the transcriptomic and chromatin landscapes of distinct EMT states in a mouse model of skin squamous cell carcinoma (SCC). Through CRISPR/Cas9-mediated loss-of-function studies coupled with in vitro and in vivo functional assays, we identify TFs regulating specific EMT states. Klf5 and Pitx1 control the early stages of EMT and are essential for metastasis formation. In contrast, Nfatc1 and Creb3l1 act at later stages of EMT. Similar EMT states and regulatory patterns are found in mouse pancreatic adenocarcinoma and human cancers. Altogether, our study defines the transcriptional and chromatin landscape controlling EMT progression in mouse skin SCC, identifies EMT state-specific TFs and highlights their essential roles in regulating metastasis.

Tumor-infiltrating lymphocyte (TIL) therapy, which involves extracting, expanding, and reinfusing immune cells to target cancer cells, has shown promise in melanoma treatment, but requires optimization for broader efficacy. The success of TIL therapy depends on the recognition of tumor-associated antigens, but neoantigen-reactive T-cells are often rare and exhausted in less immunogenic malignancies. Isolating T cells enriched in neoantigen reactivity prior to in vitro expansion and reinfusion may improve the response rates. To this end, our proprietary Specific Neo-Antigen Peptides (SNAP™) technology platform improves the accuracy of neoantigen prediction and validation by combining advanced computational modelling and PepSeq, a high-throughput screen for the physical credentialing of putative neoantigens based on their affinity to bind patient-specific HLA class II proteins. This approach allows for the education and enrichment of TILs (SNAP-TILs) with personalized, predefined, highly immunogenic neoantigens prior to expansion. Using the SNAP platform, we consistently achieved, on average, a SNAP-TIL product comprising 96% CD3+ cells, with a mixture of 75% effector and 23% central memory cells. SNAP-TILs exhibited greater efficacy and selectivity in immune infiltration than TIL, which was expanded by the rapid expansion protocol alone using ex vivo models. SNAP-TIL was also reactive in highly and poorly immunogenic tumors, with 70% and 50% tumor growth inhibition in melanoma and pancreatic patient-derived xenograft models, respectively. This study demonstrates the novel benefit of our Personalized Neoantigen Pipeline approach, potentially providing a durable antitumor immune response for a larger proportion of cancer patients.

Tumor-seeking fluorescent dyes enable precise lesion localization by recognizing overexpressed receptors, providing a critical adjunctive technology for cancer histopathology. However, tumor heterogeneity and the poor understanding of targeting mechanisms limit their efficacy. Here we engineer CETIF6a, a click chemistry-compatible heptamethine cyanine dye, for multi-cancer targeting, intraoperative histopathology, and proteome-wide target identification. CETIF6a demonstrates margin delineation across multiple cancer types ( > 90% concordance with H&E staining). Quantitative proteomics reveals that the dye targets 5-15 times more tumor-specific proteins than in paracancerous tissues. Synergistic pan-cancer targeting is achieved through 491 conserved tumor-enriched proteins involved in ribosomal, proteasomal, and metabolic pathways, effectively overcoming heterogeneity. Mechanistic studies confirm that CETIF6a emits bright fluorescence upon covalent binding to targets via nucleophilic substitution at cysteine thiol residues within hydrophobic cavities. The modifiable scaffold of CETIF6a supports both intraoperative tumor diagnosis and functional targets profiling, providing a foundation for systematic probe optimization.