P0087 Targeting TRPM8 as a novel strategy to mitigate intestinal fibrosis in Crohn’s Disease.

Abstract Background Intestinal fibrosis is a major complication of inflammatory bowel disease (IBD), driven by aberrant activation of intestinal fibroblasts and excessive extracellular matrix deposition, ultimately leading to stricture formation (Rieder et al., 2024). Although the ion channel TRPM8 is known to modulate inflammatory pathways, including within the gut (Cicia et al., 2025), its role in intestinal fibrogenesis remains unclear. This study aims to investigate the functional role of TRPM8 in intestinal fibrosis. Methods Human intestinal biopsies from patients with fibrotic Crohn’s disease (CD) were analyzed to quantify TRPM8 expression in fibrotic versus non-fibrotic tissue specimens. Human primary intestinal fibroblasts from fibrotic CD patients were phenotypically characterized by RT-PCR for key fibrotic markers and exposed to the TRPM8 antagonist AMTB to assess proliferation. A TNBS-induced murine model of intestinal fibrosis was established, and mice were treated with AMTB (10 mg/kg). Gut fibrosis progression was evaluated by colonoscopy and assessed by qRT-PCR and histological analyses. Immune signaling was characterized by flow cytometry analysis of CX3CR1+ cells. Primary colonic fibroblasts were isolated from TNBS-treated mice receiving AMTB, and gene expression of TRPM8 and markers involved in inflammation, fibroblast activation, and matrix remodeling was assessed by qRT-PCR. Results TRPM8 expression was significantly upregulated in human fibrotic biopsies from CD patients compared to the healthy counterparts. Primary human intestinal fibroblasts displayed a profibrotic transcriptional signature and exhibited altered proliferative behavior in response to AMTB treatment. In vivo, AMTB (10 mg/kg) treatment reduced intestinal fibrosis, as indicated by colon weight/length ratio, MEICS score, and histological evaluation. Flow cytometric analysis showed AMTB-dependent modulation of CX3CR1+ immune cell populations in fibrotic colon. Primary colonic fibroblasts isolated from AMTB-treated fibrotic mice reproduced these in vivo effects, exhibiting reduced expression of inflammatory, fibroblast-activation, and matrix-remodeling markers compared to TNBS-treated fibroblasts. Conclusion Collectively, these findings identify TRPM8 as a regulator of fibroblast behavior and intestinal fibrogenesis. Pharmacological inhibition of TRPM8 with AMTB mitigates fibrotic progression in vivo and modulates both fibroblast activity and immune cell responses. TRPM8 emerges as a potential target for intestinal fibrosis. References: Rieder F, Mukherjee PK, Massey WJ, Wang Y, Fiocchi C. Fibrosis in IBD: from pathogenesis to therapeutic targets. Gut. 2024 Apr 5;73(5):854-866. Cicia D, Biscu F, Iannotti FA, Miraglia M, Ferrante C, Iaccarino N, Cadenas de Miguel S, Chiavaroli A, Schiano Moriello A, De Cicco P, Nanì MF, Zanoletti L, Ke BJ, van Baarle L, Talavera K, Randazzo A, Elia I, Capasso R, Matteoli G, Pagano E, Izzo AA. Dietary targeting of TRPM8 rewires macrophage immunometabolism reducing colitis severity. Cell Death Dis. 2025 Apr 25;16(1):343. Conflict of interest: Nani, Mariafrancesca: No conflict of interest Rinaldi, Maria Michela: No conflict of interest Miraglia, Maria: No conflict of interest De Cicco, Paola: No conflict of interest Cerqua, Ida: No conflict of interest Feliciello, Fabiana: No conflict of interest Sergio, Claudia: No conflict of interest Orsi, Mariagrazia: No conflict of interest Pagano, Ester: No conflict of interest Izzo, Angelo Antonio: No conflict of interest Romano, Barbara: No conflict of interest