The dual roles of immunomodulation and epithelial regeneration in MSCs primed by injured colonoids: a novel priming method for enhancing mucosal healing in IBD

Mucosal healing has become a critical therapeutic target, resulting in a favorable prognosis for patients with inflammatory bowel disease (IBD). The intestinal mesenchymal cells (iMCs), including mesenchymal stem cells (MSCs), play a pivotal role during mucosal healing by modulating immune response and supporting tissue repair, making them a promising target for therapeutic interventions aimed at mucosal healing in IBD. Recent advancements in priming strategies have enhanced the therapeutic potential of MSCs, primarily focused on immunomodulation. Although epithelial regeneration is also identified as a critical therapeutic target for mucosal healing in IBDs, there are currently no priming methods that specifically target the pro-regenerative properties of MSCs. Thus, we aimed to develop novel priming methods to enhance both immunomodulation and epithelial regeneration in MSCs. We, thus, hypothesize that human bone marrow-derived mesenchymal cells (HBMMSC) could directly communicate with injured human colonoids and be primed to enhance the secretion of immunomodulatory and pro-regenerative factors. We employed coculture models to investigate the molecular response of HBMMSC to colonoid, both under normal conditions and following colonoid injury. This injury was induced by an IFN-γ and TNF-α cocktail in anoxic conditions, simulating intestinal inflammatory injury. To better understand the HBMMSC whole transcriptomic response to injured colonoids, we performed RNA sequencing for HBMMSC cocultured with normal or injured colonoids and HBMMSC-only groups. In a co-culture setting under normal conditions, we observed HBMMSC support colonoid spheroidal growth. Pro-WNT ligand genes ( WNT2B, WNT4, WNT5A, and RSPO3 ) were enhanced in the co-cultured HBMMSC with normal colonoids compared to the HBMMSC-only group. Consistent with previous reports on iMC response to epithelial damage, HBMMSC cocultured with injured colonoids substantially expressed the pro-WNT signaling genes when compared with HBMMSC cocultured with normal colonoids, suggesting their pro-regenerative support for colonoid injury repair processes. The transcriptomic data identified significant differentially expressed genesets (DEGs) (q-value ≤ 0.05). Interestingly, we found upregulated genes related to chemokines (CXCL11, 10, 9 and CCL5, 8, etc.), anti-inflammatory cytokines (IDO1 and TGF-b, etc.), and WNT signaling amplifiers (a 3.64fold change of RSPO3 and a 1.83fold change of WNT5A) in HBMMSC cocultured with injured colonoids against the HBMMSC cocultured with normal colonoids. Gene ontology (GO) and KEGG pathway analyses suggest potential crosstalk between MAPK and WNT signaling mediators. The results show that HBMMSC primed by damaged colonoids enhances cellular signaling for immunomodulation and tissue repair. The findings from this study may unveil innovative priming methods for MSC-based therapies aimed at improving mucosal healing in those with IBD by boosting both immunomodulation and epithelial regeneration. The authors appreciate the financial support from the MAES Competitive Grant Program at the University of Maryland, College Park, USA. This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.