Integrating network pharmacology and metabolomics to explore the mechanism of Xihuang Pill in exerting anti-tumor immune effects against pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is driven by genetic alterations in the pancreatic epithelium (e.g., KRAS) coupled with dysregulated innate immunity that triggers tumor-promoting chronic inflammation. However, the identity of innate immune molecular regulators as therapeutic targets in PDAC is ill-defined. Here, we show in PDAC patients that elevated tumoral expression of the inflammasome adaptor protein ASC and its downstream effector Caspase-1 is primarily colocalized to the pancreatic ductal epithelium and prognostic for poor survival. In the mutant Kras-driven KPC PDAC mouse model, global and conditional (pancreatic epithelial) ablation of ASC, or nanobody-mediated targeting of extracellular ASC, suppresses pancreatic tumorigenesis. Whole transcriptome profiling and multiplex immunofluorescence reveal that the tumor-promoting activities of epithelial-derived ASC align with molecular pathways for mitochondrial respiration, metabolism (glycolysis), and immune responses. Our discovery that ASC-containing inflammasomes promote PDAC by acting as a molecular bridge between innate immunity, mitochondrial dysfunction and metabolic reprogramming provides the rationale to therapeutically target ASC in cancers.

GATA6 promotes epithelial phenotypes and limits epithelial-to-mesenchymal (EMT) transition in pancreatic ductal adenocarcinoma (PDAC). Here we show that GATA6 defines a tumor cell state that induces MHCI expression and anti-tumor cytotoxicity upon therapy. In human PDAC, GATA6 expression correlates with immune cell infiltration, and spatial analysis reveals interaction between GATA6+ tumor cells and CD8+ T cells. In murine PDAC, MEK inhibition (MEKi) enriches antigenicity-related gene sets in GATA6high cells, while GATA6 knockout or degradation impairs MEKi-induced MHCI upregulation. High-GATA6 tumors respond to MEKi with increased MHCI, enhancing T-cell cytotoxicity, whereas GATA6 loss abolishes this effect. Treatment-induced EMT reduces GATA6+ populations and MHCI expression, which is restored by combining MEKi with HDAC inhibitors, enhancing GATA6+ tumor cells, MHCI, CD8+ T cell infiltration, tumor suppression, and survival. These findings suggest that therapeutic strategies promoting a GATA6-driven tumor cell state improve immune recognition of PDAC cells and potentiate anti-tumor cytotoxic effects.

Pancreatic ductal adenocarcinoma (PDAC) remains a deadly malignancy owing to its late presentation and the limited sensitivity of current serum biomarkers and imaging for early detection. Extracellular vesicles (EVs), which carry proteins, nucleic acids, and lipids that reflect tumor-stromal interactions, have emerged as promising biomarkers for early diagnosis, disease monitoring, and treatment response. Several EV-derived proteins, microRNAs, long noncoding RNAs, and DNA alterations linked to early carcinogenesis and therapeutic resistance have been identified. However, variability in pre-analytical handling and analytical platforms has hindered reproducibility. Global standardization efforts, such as European Liquid Biopsy Society (ELBS), The Blood Profiling Atlas in Cancer (BloodPAC), International Liquid Biopsy Standardization Alliance (ILSA), and Minimal information for studies of extracellular vesicles (MISEV), are currently helping to unify methodologies for EV isolation and molecular profiling. Advances in analytical technologies have shifted the field from bulk EV measurements to high-resolution single-vesicle approaches. Techniques, such as nanoflow cytometry, super-resolution imaging, Raman spectroscopy, and surface-enhanced Raman scattering, enable the detection of rare, mutation-bearing, or functionally distinct EV subpopulations, which may enhance diagnostic precision. In gastroenterology, a major opportunity lies in integrating single-EV analytics with the endoscopic sampling of tumor-proximal fluids. EVs obtained from pancreatic juice, bile, duodenal fluid, or portal venous blood via endoscopic retrograde cholangiopancreatography (ERCP) or endoscopic ultrasound (EUS) provide spatially enriched molecular information beyond peripheral blood. Combining endoscopic access with particle-level EV characterization may allow real-time, mechanism-informed assessment of tumor biology and premalignant lesions, offering a promising strategy for early detection and risk stratification in PDAC. Together, these developments have positioned EV-based liquid biopsy as a rapidly maturing field with strong translational potential.

Mutant p53(Mtp53) not only loses its canonical tumor-suppressive functions but also acquires oncogenic gain-of-function properties, positioning it as a central orchestrator in reshaping the tumor immune microenvironment. This review systematically delineates how Mtp53 actively establishes and sustains an immunosuppressive niche through multiple interconnected mechanisms, including chronic inflammation, immune cell dysfunction, reprogramming of cancer-associated fibroblasts, metabolic dysregulation, epigenetic hijacking, and potentially aberrant liquid–liquid phase separation, thereby promoting immune evasion and therapeutic resistance. We integrate current evidence to propose a conceptual “metabolism–epigenetics–immunity” axis: Mtp53-driven metabolic reprogramming—such as accumulation of lactate or α-ketoglutarate—can modulate chromatin modifications and immune gene expression. Notably, the full in vivo causal chain of this axis remains unestablished; existing support derives primarily from stepwise experimental data and strong correlations. The immunological impact of Mtp53 is highly context-dependent, shaped by co-mutations and tissue origin. In TP53/KRAS co-mutant non-small cell lung cancer (NSCLC), Mtp53 enhances tumor immunogenicity and improves response to immune checkpoint inhibitors (ICIs); conversely, in immunologically “cold” tumors—such as triple-negative breast cancer, pancreatic ductal adenocarcinoma, and colorectal cancer—it promotes T-cell exhaustion or myeloid suppression, reflecting marked cancer-type heterogeneity. Therapeutic approaches include Mtp53 reactivators (e.g., APR-246, PC14586), degraders, synthetic lethal strategies, and neoantigen vaccines. Although APR-246 showed efficacy in a phase II trial (NCT03072043), it failed to improve survival in phase III (NCT03745716) due to lack of TP53 mutation stratification. Its combination with pembrolizumab (NCT04383938) demonstrated acceptable safety (immune-related adverse events in ∼12%) but limited efficacy, underscoring the need for biomarker-guided, precision-based combinations. Thus, a multidimensional biomarker platform is urgently needed—one integrating TP53 mutation subtypes (e.g., R175H vs . nonsense mutations), dynamic ctDNA monitoring (VAF ≥ 0.01%), tumor immune microenvironment (TIME) features (e.g., TILs, MDSCs), and spatial multi-omics—to enable precise molecular stratification and personalized intervention in Mtp53-driven cancers.

Circulating tumor DNA-guided early detection and minimal residual disease monitoring in pancreatic and biliary cancers: evidence, barriers, and opportunities.

Pancreatic ductal adenocarcinoma (PDAC) cells undergo mitochondrial metabolic reprogramming to support their proliferation. However, the mechanisms by which mitochondrial protein quality control (MPQC) regulates cell metabolism remain unclear. Here, we found that c-Myc promotes PDAC cell proliferation by transcriptionally upregulating the expression of GRPEL1, an essential MPQC component. Mechanistically, c-Myc-regulated GRPEL1 maintains oxidative phosphorylation (OXPHOS) and minimizes ROS accumulation, thereby facilitating de novo fatty acid (FA) synthesis through the transcriptional upregulation of fatty acid synthase (FASN) expression. Targeting the c-Myc/GRPEL1 axis to block FASN-regulated FA synthesis inhibited PDAC cell proliferation and tumor growth in both cell models and patient-derived organoids (PDOs), whereas FA supplementation partially reversed this inhibitory effect. Clinically, c-Myc expression is positively associated with the levels of MPQC components in pancreatic ductal cells, with GRPEL1 ranking among the top hits. Furthermore, c-Myc, GRPEL1, and FASN are all expressed at higher levels in PDAC tissues than in peri-tumoral pancreatic tissues, and both c-Myc and GRPEL1 expression levels are positively correlated with that of FASN. These findings suggest that therapeutic inhibition of FA synthesis may be promising for treating PDAC patients with active c-Myc/GRPEL1/FASN signaling. Overall, this study demonstrates that FA synthesis mediated by the c-Myc/GRPEL1/FASN axis is essential for PDAC growth.

Pancreatic cancer remains one of the most aggressive digestive neoplasms, especially due to late diagnosis. The aim of our study was to investigate cytokeratin-19 fragments (CYFRA 21-1), osteopontin (OPN), and human epididymis protein 4 (HE4) clinical significance in pancreatic adenocarcinoma. Our research is a single-center cross-sectional prospective study that included sixty hospitalized patients diagnosed with pancreatic adenocarcinoma and fourteen controls. CYFRA 21-1, OPN, and HE4 were tested in all participants using Luminex x MAP technology. Serum CYFRA 21-1 levels were weakly correlated with those of OPN (r = 0.302; p = 0.009), HE4 (r = 0.485; p < 0.001), and carbohydrate antigen (CA) 125 (r = 0.376; p = 0.037). Similarly to CA 19-9 and CA 125, the serum OPN levels were higher in patients with pancreatic cancer when compared to controls, 3.37 (1.84; 9.12) ng/mL versus 1.59 (1.09; 2.51) ng/mL; p = 0.003. However, in multivariate analysis, the OPN was not an independent predictor for pancreatic cancer. Further, the receiver operating characteristic (ROC) curve analysis identified CA 19-9 as the biomarker with the highest diagnostic accuracy, while CYFRA 21-1, OPN, and HE4 did not reach clinically meaningful results. Further, the CYFRA 21-1 levels were significantly higher in cases subjected to significant weight loss before admission.

Pancreatic ductal adenocarcinoma (PDAC) has a relatively low incidence but a high mortality rate, primarily due to difficulties in early detection. Current state-of-the-art methods for diagnosing early-stage PDAC tend to be invasive, time-consuming, and unreliable, primarily due to the difficulties associated with the early detection of pancreatic cancers. Here we show a quick and sensitive method for the early diagnosis of PDAC using a signal-enhanced lateral flow immunoassay called SELFI. We develop SELFI, which generates a strong colorimetric signal through multiple hotspots formed by plasmonic gold nanoparticles (AuNPs) assembled on a silica nanoparticle. Our SELFI assay achieves a 10,123-fold increase in the limit of detection compared to conventional lateral flow immunoassays using 20 nm AuNPs, providing results within 15 min. We demonstrate that SELFI enables early diagnosis of PDAC, as indicated by a receiver operating characteristic curve and a larger area under the curve compared to the enzyme-linked immunosorbent assay. SELFI's effective diagnostic features can enhance the timely identification of PDAC and may also serve in the early diagnosis of a range of other diseases.

Hypofractionated radiotherapy (HFRT) is increasingly used not only for local tumour control but also for its immunomodulatory potential. By delivering higher doses per fraction over fewer sessions, HFRT improves local disease control and reshapes the tumour immune microenvironment. This review integrates preclinical, translational, and clinical evidence on the immunological effects of HFRT when combined with immune checkpoint inhibitors (ICIs) and other immune-based therapies. Available evidence indicates that HFRT induces immunogenic cell death and activates the cGAS-STING pathway, enhancing dendritic cell priming and CD8+ T-cell trafficking. These processes are most likely to translate into systemic antitumour activity when checkpoint blockade is delivered with an appropriate peri-radiotherapy window. Under these conditions, HFRT may facilitate immune conversion of selected “cold” tumours, particularly in combination with PD-1/PD-L1 blockade. Clinical outcomes remain heterogeneous across tumour types. Improved out-of-field responses and survival signals have been reported in non-small-cell lung cancer, head and neck squamous cell carcinoma, and triple-negative breast cancer, whereas tumours dominated by myeloid-driven or stromal suppression, such as pancreatic ductal adenocarcinoma, show limited benefit. From a clinical design perspective, effective HFRT-immunotherapy combinations require careful selection of fractionation, timing, and radiation geometry. Fractionation should preserve DNA sensing and dendritic-cell activation, checkpoint therapy should align with the peri-radiotherapy window, and radiation delivery should minimise immune suppression by sparing tumour-draining lymph nodes and limiting unnecessary low-dose exposure. HFRT can serve as a practical backbone for immune-based therapies when these variables are appropriately aligned.

Di(2-ethylhexyl) phthalate (DEHP) is a ubiquitous plasticizer implicated in pancreatic carcinogenesis, yet the molecular initiating events and adverse outcome pathways (AOPs) linking exposure to disease mechanisms remain poorly resolved. In this study, we integrated a multi-scale systems toxicology framework—combining heterogeneous ensemble machine learning, Mendelian randomization (MR), molecular docking and molecular dynamics (MD), single-cell transcriptomics, and in vitro assays—to delineate a candidate mechanistic trajectory. A Tabular Prior-Data Fitted Network (TabPFN)-enhanced ensemble identified a six-gene pancreatic ductal adenocarcinoma (PDAC) signature (AUC = 0.946). Within this signature, MR provided suggestive evidence for a modest association between genetically predicted Krüppel-like factor 5 (KLF5) expression and pancreatic cancer risk (OR = 1.188, p = 0.046). Functional enrichment of DEHP–PDAC intersection targets highlighted pro-survival signaling modules, including PI3K–AKT- and MAPK-related pathways. Structure-based analyses supported the biophysical plausibility of a non-covalent DEHP–KLF5 interaction (−6.4 kcal/mol), and 100-ns MD simulations indicated a persistent binding mode with conformational accommodation. Single-cell analysis localized KLF5 predominantly to malignant ductal cells and, together with CellChat inference, was consistent with malignant cell-derived TGF-β- and MIF-related signaling. Network-based virtual knockout further suggested that KLF5 may contribute to sustaining a TGF-β/MMP7-linked matrix remodeling program. Consistently, DEHP exposure upregulated KLF5 and MMP7 and enhanced migration and invasion of PANC-1 cells. Collectively, these findings support a working AOP model linking DEHP-responsive KLF5-centered activity to extracellular matrix (ECM) remodeling and immunomodulatory communication, providing a mechanistic rationale that aligns with signaling programs characteristic of therapy-tolerant tumor niches, particularly PI3K–AKT-coupled survival signaling and TGF-β-linked stromal remodeling. Although these implications are hypothesis-generating, they highlight a potential avenue for future drug-response investigations.

The αvβ6 integrin has emerged as a valuable target for theranostic applications in nuclear medicine with high applicability across a variety of cancers, including head-and-neck, lung, breast, and pancreatic carcinomas. [⁶⁸Ga]Ga-Trivehexin is a prominent example of a diagnostic tracer targeting this integrin. In this work, we aimed to expand on this concept by developing FSC(PEG4-αvβ6)₃, a novel tracer that retains the Trivehexin design, but features PEGylated spacers and replaces the TRAP chelator with Fusarinine C (FSC), enabling labelling with Zirconium-89 in addition to Gallium-68. Preclinical characterization of [⁶⁸Ga]Ga/[⁸⁹Zr]Zr-FSC(PEG4-αvβ6)₃ included affinity determination towards the αvβ6 integrin and cellular uptake studies in αvβ6-positive H2009 cells. A subcutaneously xenografted H2009 tumor model was used to assess the PET imaging potential and biodistribution at early time points with the Gallium-68 labelled compound, and at later time points (up to 6 days post-injection) with the Zirconium-89 labelled version. While [⁶⁸Ga]Ga-FSC(PEG4-αvβ6)₃ exhibited moderate binding to αvβ6, its affinity, cellular internalization, and tumor uptake in vivo were lower compared to [⁶⁸Ga]Ga-Trivehexin. Notably, this decreased target engagement was associated with reduced nonspecific binding, which we primarily attributed to the incorporation of PEGylated linkers. Despite indication of in vivo degradation of [⁸⁹Zr]Zr-FSC(PEG4-αvβ6)₃, still a meaningful evaluation of pharmacokinetics and biodistribution at extended time points was feasible, revealing prolonged tumor persistence up to 6 days post-injection. FSC(PEG4-αvβ6)₃ represents the first-generation tracer targeting the αvβ6 integrin based on the multifunctional chelator Fusarinine C, thereby expanding the repertoire of radionuclides applicable from Gallium-68 to Zirconium-89. Following further optimization, this novel class of compounds holds significant promise for enabling clinical translation and advancing the development of next-generation αvβ6-directed imaging agents.

Identification of HSPE1 as a new actionable cancer vulnerability leads to an innovative and effective combination therapy for pancreatic ductal adenocarcinoma.

Extraordinary enlargement of the main pancreatic duct dilatation in pancreatic ductal adenocarcinoma – a case report

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy with a poor prognosis and increasing global incidence. Although the epigenetic regulator plant homeodomain finger protein 1 (PHF1) is known for its transcriptional silencing function in cancer, its role in PDAC progression and translational regulation remains largely unexplored. Here, we report that PHF1 is significantly upregulated in PDAC and promotes the proliferation, migration, and invasion of PDAC cells. Mechanistically, PHF1 physically interacts with the fibrillarin (FBL)/NOP56/NOP58 snoRNP complex, facilitating its assembly and enhancing the internal ribosome entry site (IRES)‐dependent translation of Snail. Depletion of PHF1 reduces Snail levels, leading to ferroptosis sensitization in KRAS‐mutated PDAC cells. Additionally, PHF1 knockdown markedly reduces overall tumor burden in Kras‐mutated mice. In conclusion, our findings identify PHF1 as an oncogene that promotes PDAC progression and demonstrate that its inhibition induces ferroptosis via suppression of IRES‐dependent Snail translation. Thus, our findings reveal a previously unrecognized translational regulatory role of PHF1 and suggest its targeting as a promising therapeutic strategy for PDAC.

Whether lymph node burden modifies chemotherapy effectiveness in pancreatic ductal adenocarcinoma (PDAC) has not been systematically investigated. We analyzed 22,045 surgically treated PDAC patients from the Surveillance, Epidemiology, and End Results database (2004-2022). Restricted cubic spline analysis examined chemotherapy hazard ratios across positive lymph node counts. Cox models estimated survival associations and interaction effects between N stage and chemotherapy on multiplicative and additive scales, with Fine-Gray competing risk models for sensitivity analyses. Additional sensitivity analyses included the exclusion of neoadjuvant therapy recipients, lymph node ratio (LNR) analysis, and stratification by lymph node examination adequacy. Chemotherapy absence increased disease-specific mortality (adjusted HR 1.72; 95% CI, 1.65-1.79), while N1 and N2 disease conferred additional risk (HR 1.54 and 2.05, respectively; both p < 0.001). Chemotherapy benefit increased progressively with nodal burden: hazard ratios for chemotherapy absence versus receipt were 1.52 (95% CI, 1.42-1.63) in N0, 1.80 (95% CI, 1.70-1.92) in N1, and 1.97 (95% CI, 1.83-2.12) in N2 disease. Adjusted 3-year disease-specific survival with versus without chemotherapy was 55.1% versus 39.3% (N0), 36.5% versus 18.2% (N1), and 26.4% versus 8.7% (N2). Notably, N1 patients receiving chemotherapy achieved better outcomes than N0 patients without chemotherapy. Adjusted interaction tests revealed significant multiplicative interactions (N1: 1.19; N2: 1.28) and additive interactions (RERI: N1: 0.65; N2: 1.29). These interaction patterns were validated in sensitivity analyses. Lymph node burden significantly modifies chemotherapy effectiveness in resected PDAC, with higher nodal burden patients deriving disproportionately greater benefit. These findings support risk-stratified treatment strategies incorporating lymph node status.

The tumor landscape of pancreatic ductal adenocarcinoma (PDAC) is refractory to conventional photon radiotherapy (RT) because of a fibrotic tumor microenvironment (TME) that promotes chronic hypoxia and reduced immune surveillance. The radiobiological factors unique to carbon ion RT (CIRT), such as high linear energy transfer and less dependence on oxygen, make it well-suited to overcome the PDAC TME. In this study, we utilized clonal syngeneic KPC pancreatic tumor cell lines and tumors to examine this postulate and to identify underlying factors that affect the response of PDAC to CIRT. Although KPC cell lines exhibited radiobiological effectiveness greater than 3, subcutaneous tumors in the mouse hind leg showed lower radiobiological effectiveness-1.3 based on quintupling time-at a linear energy transfer between 70 and 80 keV/μm. Four days after CIRT, we observed widespread transcriptomic changes in the tumor immune microenvironment, suggesting increased infiltration of antitumor immune cells and elevated expression of antitumor T-cell cytokines, MHC class I molecules, and co-stimulatory signals. Fewer immunologic changes were observed following photon irradiation. By 7 days after CIRT, tumor-supportive transcriptomic programs characterized by protumor cytokines, M2 macrophages, and cancer-associated fibroblasts emerged, promoting resistance and limiting the durability of tumor growth delay. These findings suggest that CIRT may offer a favorable platform compared with conventional photon RT for combining with immunotherapies. Furthermore, these data highlight the risk of using in vitro survival data alone in treatment planning and indicate that underlying TME factors affect the response of PDAC in vivo.

USP20-RAB8A signaling axis restricts pancreatic cancer progression by disrupting GLUT1 vesicular trafficking and inhibiting glucose uptake.

Immunosuppression within the tumor microenvironment (TME) is a major factor driving pancreatic cancer progression and therapeutic resistance. To address this challenge, we developed a nano-codelivery system, CGT (Cilengitide)-Cls-PTX (paclitaxel)/CM (cell membrane), for the co-delivery of PTX and tumor cell lysate-derived antigens from pancreatic cancer cells (from human pancreatic cancer PANC-1 and mouse pancreatic cancer PANC02 cells). The system was constructed by synthesizing an integrinαvβ3-targeting lipid, DSPE-PEG2000-CGT, and fusing PTX-loaded liposomes with pancreatic cancer cell membranes. This strategy enables preferential accumulation in the TME, where tumor antigens are released to stimulate dendritic cell (DC) maturation and relieve TME immunosuppression, thereby achieving synergistic antitumor efficacy via PTX-mediated tumor cell killing and antigen-induced immune activation. Physicochemical characterization by1H-nuclear magnetic resonance, transmission electron microscopy, and Western blot confirmed successful synthesis and membrane fusion. Immunostimulatory activity was evaluated using ELISA, flow cytometry, and co-culture assays, and therapeutic efficacy was assessed in a PANC02 murine pancreatic cancer model with Cls-PTX as the control. CGT-Cls-PTX/CM significantly enhanced DC maturation, upregulated co-stimulatory molecules (CD80, CD86), and promoted secretion of interleukin-6 (IL-6) and interleukin-12 (IL-12). Furthermore, it increased CD4+and CD8+T-cell proliferation, elevated interferon-γ(IFN-γ) production, suppressed transforming growth factor-β, and facilitated cytotoxic T lymphocyte infiltration into tumor tissues. Overall, CGT-Cls-PTX/CM effectively remodels the immunosuppressive TME, achieving synergistic antitumor effects through combined chemotherapy and immune modulation. This strategy offers a promising approach for enhancing immunotherapeutic efficacy against pancreatic ductal adenocarcinoma, a prototypical 'cold' tumor resistant to immune checkpoint therapy.

Frozen section (FS) tissue assessment is essential for guiding intraoperative surgical decision-making in oncology, particularly in procedures such as pancreatic ductal adenocarcinoma (PDAC) resections, where margin status critically impacts patient survival. The current gold standard, (FS), while widely used, suffers from notable limitations, including tissue artifacts, dependence on specialized expertise, and slow turnaround times, resulting in sampling errors and false negatives. To address these challenges, we present a novel approach for automatic cancer identification in fresh tissue biopsies using mul tispectral Mueller Matrix (MM) polarimetry. Our custom-built multispectral MM polarimeter captures polarization-resolved imaging across multiple wavelengths, enabling pixel-level analysis of tissue microstructure without staining or histology sectioning. Our approach thus allows for assessments in quasi-real time. From these, we propose a deep learning model that uses MM data collected from PDAC patients to distinguish cancerous from non-cancerous biopsies to assess samples automatically. Experimental results demonstrate classification performance comparable to RFS assessments performance found in clinical routine, with enhanced diagnostic speed. We show that our approach is consistent and coherent against pixel-wise annotations from histology slides. This study highlights the potential of MM polarimetry combined with machine learning as a viable, label-free alternative for real-time intraoperative cancer detection.