Introduction

Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with the lowest 5-year survival rate of all cancers due to late diagnosis and lack of effective treatment. One of the challenges of developing molecular diagnosis and molecular targeted therapy for PDAC is the “mismatch” between research models and patients.

Aims & Methods

This study aimed i) to identify common proteins among different research models and patients and accordingly ii) to identify potential hub proteins and signaling pathways involved in the PDAC pathogenesis by using proteomics-based system modeling of multimodal datasets derived from multiple research models and patients. Mouse PDAC cells (derived from a genetically engineered mouse model with KrasG12D/Pdx1-Cre), PDAC orthotopically tumors in C57B/L6 mice (n=5), mouse PDAC spheroids, mouse pancreatic organoids, mouse PDAC organoids, normal mouse pancreatic tissues (n=5), human PDAC tumor tissue samples from treatment-naїve (n=20), neoadjuvant chemotherapy (n=22), and human PDAC organoids were included. Mass spectrometry-based proteomics, Rstudio, Cytoscape 3.10 and pathway enrichment analysis (using ingenuity pathways analysis, including human PDAC transcriptomic in addition) were used.

Results

In the comparison of PDAC models and patients, shared proteins were identified as follows: 60% between the mouse PDAC cells and the mouse PDAC spheroids, 78% between the mouse PDAC cells and the mouse PDAC tissue, 56% between the mouse PDAC spheroids and the mouse PDAC tissue, 66% between the mouse PDAC cells and the mouse PDAC organoids, 54% between the mouse PDAC spheroids and the mouse PDAC organoids, 38% between the mouse PDAC and the human PDAC tissues, 31% between the human PDAC tumor and the human PDAC organoids, and 60% between the mouse and the human PDAC organoids. A total of 1,367 common proteins were identified within all the PDAC models. Among these proteins ACTB, HSP90AA1, HSPA8, HSP90AB1, HSPA4, EEF2, RPS3, RPS20 and GAPDH were the top 10 hub proteins (with >300 node connections). Upon comparison of all eight models, eight proteins were identified as unique to PDAC, namely FAHD1, ARSA, NEK9, SERPINB9, LAMTOR2, SNRPG, TMED3, and RAB1A. There was a correlation in terms of z-score regarding signaling pathways between the mouse PDAC and human PDAC (p<0.001), particularly in activations of nerve growth factor signaling, integrin signaling, and neuregulin signaling. Central hub proteins commonly found in the mouse models included vinculin (membrane-cytoskeletal protein) as one of the high connectivity nodes and β-catenin and probable ubiquitin carboxyl-terminal hydrolase FAF-X (involved in axonal growth and neuronal cell migration) as high centrality nodes. In patients, central hub proteins that were dysregulated after chemotherapy included 14-3-3 protein zeta/delta (encoded by YWHAZ gene), β-catenin and SRC. Among the 47 proteins that were associated with altered metabolism after the chemotherapy in patients, CD44, CTNND1, APOA 1, PARVA, and SCRBS1 were nodes with low degrees, and desmosomes (DSP, KP3, PPL), PRTN3, thrombospondin-2 (THBS2), and DMBT1 were the most down-regulated proteins without nodes, and APOA1, caveolin-1 (encoded by CAV1), LRP1 were the most up-regulated proteins with nodes.

Conclusion

The system modeling identified the common proteins among the research models and patients and the hub proteins and signaling pathways, suggesting the potential translational targets of PDAC.

Clinical Case Summary

A 24-year-old male presented with jaundice and mild vertigo after a 10-day prodrome (asthenia, nausea, headache, fever). Ibuprofen (1200 mg/d) was taken for these symptoms only during the first 5 days. Labs revealed acute hepatocellular injury and elevated cholestasis (AST 215, ALT 862, FA 134, GGT 126 IU/L), marked mixed hyperbilirubinemia (Total 13.5 mg/dL, Direct 7.58 mg/dL) and coagulopathy (INR 1.59). Ultrasound showed hepatosplenomegaly. Viral hepatitis (HAV/HBV/HCV/CMV) and autoimmune/metabolic causes were excluded. Epstein-Barr Virus (EBV) serology showed low-titer positive VCA IgM but negative VCA IgG, EBNA IgG, and serum EBV DNA PCR, indicating recent non-active infection. Given ibuprofen timing and high RUCAM score, drug-induced liver injury (DILI) with a hepatocellular pattern was suspected. Liver biopsy confirmed acute hepatitis with lymphomonocytic infiltrate, necrosis and canalicular cholestasis. Despite improving liver function tests post-ibuprofen cessation, he developed worsening vertigo, severe ataxia, nausea, vomiting, hypophonia, necessitating nasogastric feeding/bed confinement. Cerebrospinal fluid (CSF) showed mild mononuclear pleocytosis (16 cells/mm³); a multiplex PCR pathogen panel (including EBV) was negative. Brain MRI/EEG were normal. Post-infectious EBV acute cerebellar ataxia was diagnosed concurrently with DILI, based on timing post-recent EBV, exclusion of active neurological infection (negative CSF PCR, normal imaging), mild pleocytosis, and supported by later IgG seroconversion. IV methylprednisolone treatment, chosen for clinical severity, led to significant neurological improvement within days and eventual recovery after physiotherapy. This case highlights the significant challenges of diagnosis and management of concomitant DILI and recent EBV infection with neurological sequelae, requiring careful differentiation of liver damage patterns (hepatocellular DILI vs cholestatic EBV) and optimal management of neurological symptoms.

References

Illustrative Material References:
- Liver biopsy histology slides (highlighting lymphomonocytic infiltrate, necrosis, and canalicular cholestasis characteristic of DILI).
- Table/Graph showing serial Liver Function Tests (AST, ALT, Bilirubin) over time, depicting injury peak and recovery trend.
- Table summarizing sequential Epstein-Barr Virus (EBV) serology results (VCA IgM, VCA IgG, EBNA IgG) demonstrating seroconversion.
- Table summarizing Cerebrospinal fluid (CSF) analysis results (cell count, protein, glucose, negative multiplex PCR).
- Representative Brain MRI images (axial/sagittal views) confirming absence of structural lesions or encephalitis signs.

Introduction

An increased risk of pancreatic cancer has been reported in families with breast cancer history carrying BRCA1 and BRCA2 mutations. Pancreatic cancer is associated with familial and hereditary cancer syndromes. Multigene panel testing for pancreatic cancer detected germline variants BRCA1/2, PALB2, ATM, TP53, MLH1, STK11/LKB1, APC, CDKN2A, and SPINK1/PRSS1 as high-risk genes. Active pancreatic surveillance using computed tomography (CT), magnetic resonance imaging (MRI), and echoendoscopic (EE) are recommended for high-risk individuals who have a family history of pancreatic cancer or harbor these pathogenic germline variants to improve the detection rate and the prognosis of pancreatic cancer. We evaluated the performance characteristics of echoendoscopically guided fine needle biopsy (EE-FNB) in this subset of patients.

Aims & Methods

This is a retrospective analysis of a prospective database total of 99 families with ≥1 case of breast cancer tested for mutations in BRCA1 and/or BRCA2. The eligible patients were the ones that underwent EE-FNB at a tertiary hospital with an Oncology center due a focal pancreatic lesion observed on CT/MRI, from January 2022 to January 2024. Patients were excluded if (1) they had obstructive jaundice or (2) the lesion appeared cystic on CT/MRI. The main outcome measures were (1) prevalence of the association between BRCA 2 positive and pancreatic cancer (2) performance characteristics of EE-FNB for identification of malignancy.

Results

In the 99 patients in the study, a focal pancreatic lesion was identified with pancreatic adenocarcinoma by EE-FNB. The final diagnosis included 20 patients with breast cancer and BRCA2 positive undergoing oncological treatment or follow-up, with a mean age of 32.8 +/- 2.2 years. EE was performed in an average of 19 +/- 11.3 days. EE-FNB showed an accuracy of 100% for the diagnosis of malignant neoplasia, with a sensitivity of 100%, specificity of 100%.

Conclusion

EE-FNB diagnostic yield was substantial in established high-risk mutation carriers, is highly accurate in patients with focal pancreatic lesion on CT/MRI, without obstructive jaundice. EE-FNB can potentially be used as a definitive diagnostic test in the management of these patients.

Introduction

Growing evidence supports the role of environmental factors, such as diet and lifestyle, in modulating the intestinal microbiota and potentially influencing the symptoms of chronic gastrointestinal disorders. This study aimed to correlate lifestyle habits with colonic microbiota composition in patients with Inflammatory Bowel Disease (IBD) or Irritable Bowel Syndrome (IBS), compared to healthy controls (HCs).

Aims & Methods

38 IBD, 44 IBS and 48 HC were recruited. Fecal microbiota composition was analyzed by sequencing the 16S rDNA V3/V4 region using the Illumina platform. Dietary habits and physical activity levels were assessed using a 7-day food diary, a Food Frequency Questionnaire, and the IPAQ. Urinary indican was measured via colorimetric assay and HPLC, fecal zonulin levels were assessed using ELISA, and selected circulating microRNAs were analyzed in plasma and serum by TaqMan-based quantitative Real-Time PCR.

Results

A significant reduction in dietary fiber intake was observed in both IBD and IBS patients compared to healthy controls (HCs) (p < 0.05). IBS patients engaged in more moderate physical activity than HCs (p < 0.05). Reduced fiber intake was associated with decreased microbial diversity (p < 0.005), compared to adequate fiber consumption. Microbiota analysis revealed a reduction in alpha diversity in IBD patients compared to HCs, particularly in terms of phylogenetic diversity and observed features, while IBS patients showed intermediate values. Beta diversity (weighted and unweighted UniFrac) demonstrated a clear separation among the three groups, with the most pronounced differences observed between IBD and the other conditions. LEfSe analysis identified the highest number of discriminant taxa in HCs, consistent with their greater microbial biodiversity. No clearly pathogenic organisms were identified in either IBD or IBS patients, suggesting that the observed alterations may result from a loss of beneficial commensals rather than the presence of harmful species. Additionally, greater microbial diversity was found in individuals living in rural areas compared to those in urban environments. IBD and IBS patients not adhering to the Mediterranean Diet (MD) showed significantly higher zonulin levels than those following the MD (p < 0.001 and p = 0.003, respectively). Finally, miRNAs involved in inflammatory and epithelial permeability pathways were differentially expressed in both IBD and IBS patients. MiR-24 was significantly dysregulated in IBD (p = 0.01), while miR-923 also showed a significant difference in the same group (p = 0.05). MiR-221 exhibited a non-significant trend toward upregulation in IBS. These findings suggest a potential role for these miRNAs in the molecular mechanisms underlying inflammatory and functional bowel disorders.

Conclusion

The preliminary results of the present study underlined the impact of lifestyle factors, particularly diet and physical activity, on intestinal microbiota composition and barrier function in IBD and IBS. Microbial and molecular markers may support a more personalized approach to these disorders.

References

1. Alfaifi, J. et al; Deep Dive Into MicroRNAs in Inflammatory Bowel Disease. Inflamm. Bowel. Dis. 2023
2. Cena, H. et al; Biodiversity and planetary health: A call for integrated action. Lancet 2024
3. Giammona A et Al: Chronic Gastrointestinal Disorders and miRNA-Associated Disease: An Up-to-Date. Int J Mol Sci 2025
4. Ventham, N.T.et al.; Integrative epigenome-wide analysis demonstrates that DNA methylation may mediate genetic risk in inflammatory bowel disease. Nat. Commun. 2016

Introduction

Thiopurines are vital therapeutic agents for inflammatory bowel diseases but often cause severe myelosuppression. The NUDT15 polymorphism, particularly p.Arg139Cys (NUDT15R139C), is strongly linked to severe leukocytopenia and is prevalent in Asian populations. Despite thiopurine use during pregnancy, it remains unclear if NUDT15R139C affects fetal thiopurine sensitivity.

Aims & Methods

The human and murine NUDT15 proteins share 89% homology with the preservation of arginine at codon 139 of human NUDT15 in codon 138 of murine Nudt15. We established a knock-in mouse model with a point mutation (c.412C>T) generating p.Arg138Cys (Nudt15^R138C), mimicking the human phenotype. In the current study, we investigated offspring generated from Nudt15^+/R138C or Nudt15^+/+ female mice that were mated with Nudt15^+/R138C male mice and simultaneously administered with mercaptopurine (MP).To evaluate the effects of thiopurine on the fetus, we evaluated the number of hematopoietic stem cells in the fetal liver, which is the center of fetal hematopoiesis.

Results

In administration with the tolerable MP dose (0.5 mg/kg) for adult Nudt15^+/R138C mice, genotypes of the genotypes of neonatal mice born from breeding between Nudt15^+/R138 male mice and Nudt15^+/R138 female mice were 35 (37.2%) Nudt15^+/+, 59 (62.8%) Nudt15^+/R138C, and 0 Nudt15 ^R138C/R138C. On embryonic day 14.5, Nudt15^R138C/R138C fetal mice showed shorter body lengths and fewer hematopoietic stem cells in the fetal liver than Nudt15^+/+ fetal mice. The number of fetal HSCs was not altered in any of the Nudt15genotypes without MP treatment. However, it was significantly reduced by MP treatment in Nudt15^+/R138C fetal mice (p< 0.01, mean number 1104 in DMSO and 577 in MP) and in Nudt15^R138C/R138C fetal mice (p< 0.01, mean number 927 in DMSO and 402 in MP), but not in Nudt15^+/+ fetal mice (p= 0.39, mean number 1004 in DMSO and 820 in MP). The number of HSCs in Nudt15^R138C/R138C fetal livers was significantly reduced to less than 50% of that in Nudt15^+/+ fetal livers by MP treatment during gestation (p= 0.0089). In administration with the tolerable MP dose (1.0 mg/kg) for Nudt15^+/+ adult mice, genotypes of neonatal mice born from breeding between Nudt15^+/R138 male mice and Nudt15^{++ /}female mice were 23 Nudt15^+/+ and 0 Nudt15^+/R138C. Without MP administration, neonatal mice were born in a Mendelian fashion.

Conclusion

Fetal toxicity may arise if the fetus has more NUDT15 R139C alleles than the mother, despite safe thiopurine doses for the mother. Analysis of the father's NUDT15 genotype may enable safer thiopurine use during pregnancy, emphasizing the importance of considering parental genotypes in therapeutic decisions.