Introduction

Acute pancreatitis (AP) is an acute inflammatory autodigestive disorder triggered by dysfunction of pancreatic exocrine activity and represents the most common gastrointestinal emergency in clinical practice. Global data indicate an annual incidence rate of 33.74 per 100,000 population and a mortality rate of 1.16 per 100,000. Despite ongoing optimization of therapeutic strategies, current clinical management primarily relies on supportive care and surgery. Hypertriglyceridemia (HTG) is the second leading etiological factor for AP, accounting for 10.36% of cases. Compared to other causes, HTG-associated acute pancreatitis (HTG-AP) exhibits higher recurrence rates and more severe complications (e.g., pancreatic necrosis and renal insufficiency). With the rising prevalence of HTG, the disease burden of HTG-AP is increasing, yet its specific pathogenic mechanisms remain unclear.

Aims & Methods

This study integrated whole-genome sequencing (WGS) and next-generation sequencing (NGS) to screen for shared mutations in the exonic regions of 5 HTG-AP family members. Candidate genes were identified based on MutationTaster classification and CADD scores. A mouse model carrying the PDLIM2 gene rs145349678 mutation was constructed using CRISPR/Cas9 technology, and an HTG-AP animal model was established. Pancreatic and hepatic histopathology (HE staining) and serological markers (TG, lipase, amylase, IL-6) were analyzed to evaluate the impact of the mutation on pancreatitis severity and hepatic lipid deposition.

Results

A clinical HTG-AP family was identified, with all 5 members exhibiting hypertriglyceridemia, including 2 cases of recurrent pancreatitis. WGS revealed 278 mutation sites across 10 chromosomes (involving 23 genes) in the family, and NGS validation narrowed these to 7 candidate loci. The PDLIM2 rs145349678 mutation (NM_001368120.1:exon9:c.C1055T:p.A352V), located within the LIM domain (which mediates protein localization and signaling transduction), was selected as the target due to cross-species conservation. Experimental results demonstrated that HTG-AP mice with the PDLIM2 mutation exhibited significantly aggravated pancreatic pathological damage (edema, hemorrhage, and inflammatory cell infiltration; *P* < 0.05), elevated serological markers (TG: 2.3 ± 0.4 vs. 1.5 ± 0.3 mmol/L; lipase: 520 ± 68 vs. 320 ± 45 U/L; IL-6: 48.2 ± 6.1 vs. 28.5 ± 4.3 pg/mL; *P* < 0.01 for all), and a 2.3-fold increase in hepatic lipid deposition (*P* < 0.001) compared to controls.

Conclusion

The PDLIM2 rs145349678 mutation exacerbates inflammatory responses and hepatic lipid accumulation in HTG-AP by modulating lipid metabolism. This finding provides new insights into the pathogenesis of HTG-AP and potential therapeutic targets.

Introduction

Inflammatory bowel disease (IBD) is a growing global health problem with increasing prevalence. Current therapeutic strategies for IBD are mainly aimed at modulating the dysregulated immune activity in the intestinal mucosa. Targeting the JAK/STAT signalling pathway has been shown to be highly effective in ulcerative colitis, but not in Crohn's disease (CD), where efficacy was found to be insufficient in the initiation phase of a clinical trial [1]. Failure of the therapeutic response in Crohn’s disease may be influenced by the unique immune microenvironments in the small and large intestine and their characteristic cytokine profiles and immune cell compositions.

Aims & Methods

To gain deeper insight into region-specific efficacy in the inflamed ileum versus colon, this study investigates the immunomodulatory effects of filgotinib, a Jak1 inhibitor, using precision-cut intestinal slices (PCIS) from patients with Crohn's diseases. PCIS are viable, ultra-thin human tissue slices that retain the original intestinal architecture ex vivo and contain important cell types, including resident immune cells.
PCIS were prepared from resected ileal and colonic tissue of CD patients. The slices were cultured ex vivo and stimulated with concanavalin A or anti-CD3/CD28 antibodies to induce T cell-driven inflammatory responses. Filgotinib (1-100 µM) was applied to assess the immunomodulatory effects of JAK1 inhibition following Concanavalin A preactivation. To exclude treatment-related toxicity, cellular integrity was evaluated using an LDH release assay. Tissue morphology was examined by H&E staining and immunofluorescence. The immune response was assessed by multiplex ELISA of 20 cytokines.

Results

After filgotinib treatment, primary tissue slices from the ileum and colon of CD patients remained viable. Regional morphological characteristics of ileum and colon tissue were preserved after ex vivo cultivation, with prominent villi and smaller crypts in the ileum, and larger, more frequent crypts and flattened villi in the colon. Compared to non-CD tissue slices, Crohn’s disease-related changes, such as immune cell accumulation, were also observed in CD tissue slices ex vivo. Upon T cell-specific stimulation with anti-CD3/CD28 antibodies, region-specific immune reactivity was observed. Colonic PCIS showed an increased release of IL-2 (~3-fold), IL-10 (~2‑fold) and TRAIL (~4-fold) compared to ileal PCIS, indicating a stronger involvement of Th and regulatory T cells in the colon. In contrast, ileal PCIS showed higher levels of MIP-3α (2-fold) and granzyme A (~8-fold) after stimulation, suggesting increased cytotoxic T cell activity in this region. Filgotinib treatment showed region-specific differences in its ability to modulate cytokine release in PCIS of CD patients. In colonic PCIS, filgotinib reduced the induced secretion of Th17-associated cytokines such as IL-17F and IL-22 by up to 95%, whereas in ileal PCIS it showed no effect on the same cytokines.

Conclusion

In conclusion, filgotinib exhibited more potent anti‑inflammatory effect on Th17 cells in the colon, and a markedly reduced efficacy in the ileum. This study demonstrates for the first time that precision-cut intestinal slices can be used to predict region-specific drug response in inflamed intestinal tissue ex vivo, underscoring the critical importance of region-specific immune profiling in IBD and highlight the limited efficacy of JAK1 inhibition in ileum. This ex vivo, patient-derived model represents a valuable platform for evaluating localised treatment efficacy and advancing personalised therapeutic strategies in IBD.

References

[1] Rogler G. Efficacy of JAK inhibitors in Crohn's Disease. J Crohns Colitis. 2020 Aug 1;14(Supplement_2):S746-S754. doi: 10.1093/ecco-jcc/jjz186