Obesity-Driven Fibroblast Remodelling in Intestinal Fibrosis: A Multi-Omic Approach

Background and aims Intestinal fibrosis is a major complication of Inflammatory Bowel Disease, contributing to morbidity, surgical interventions, and healthcare burden. The rising prevalence of obesity further complicates this scenario, as obesity promotes fibrotic disease progression through inflammatory and metabolic dysregulation. Due to the heterogeneity of fibrosis and ineffective treatments, a deeper understanding of fibroblast subpopulations is essential for developing targeted therapies. This project addresses this need by exploring a novel lipid signalling pathway involving N-acyl phosphatidylethanolamine phospholipase D (NAPE-PLD) enzyme and its downstream Acylethanolamides, focusing on intestinal fibroblasts under obesogenic and chronic inflammatory conditions. Methods We will employ an integrated in vivo and multi-omics approach. Mice with intestinal epithelial-specific deletion of NAPE-PLD (NapepldΔIEC) will be exposed to chronic TNBS-induced inflammation under standard or high-fat diets. Fibrotic progression will be assessed through endoscopy, histology, and molecular profiling. Fibroblast heterogeneity will be explored using single-cell RNA sequencing to identify transcriptional changes related to fibrosis and metabolic stress. CosMx Spatial Molecular Imaging will provide high-resolution spatial maps of fibroblast–immune interactions in fibrotic tissue. Flow cytometry will validate fibroblast subset’s identity and activation states and their interactions with immune cells under different metabolic and inflammatory settings. Anticipated impact We expect to: 1. Show that NAPE-PLD deletion in intestinal epithelium worsens fibrosis under inflammatory and high-fat diet conditions. 2. Identify fibroblast subtypes linked to fibrosis and metabolic imbalance, revealing how NAPE-PLD shapes their transcriptional identity. 3. Map how obesity and NAPE-PLD loss affect stromal structure and immune cell interactions via spatial transcriptomics. 4. Define immune-metabolic and extracellular matrix (ECM) profiles controlled by NAPE-PLD, uncovering potential therapeutic targets. Together, these findings will offer new insights into how lipid signalling interfaces with fibroblast biology and metabolic inflammation to drive intestinal fibrosis. This knowledge could guide the development of personalized, mechanism-based strategies for IBD-associated fibrosis.