Exploring E-Cadherin as a Therapeutic Target for Enhancing Mucosal Healing through Intestinal Stem Cell Regeneration

Despite recent advances in treatment in patients with inflammatory bowel diseases (IBDs), achieving mucosal healing remains challenging due to an incomplete understanding of disease pathobiology and heterogeneity. Intestinal stem cells (ISCs) are crucial for epithelial regeneration and repair, making them promising targets for regenerative therapies. E-cadherin, a transmembrane protein of the adherens junctions, plays a critical role in maintaining intestinal epithelial integrity. E-cadherin modulates key signaling pathways, including Wnt, Hippo, and Notch signaling pathways, which are essential for ISC homeostasis and regeneration. However, the role of E-cadherin in ISC function and its potential as a therapeutic target for IBD have been largely unexplored. In this study, we hypothesized that downregulation of E-cadherin affects colonic stem cell development and promotes regenerative responses by modulating Wnt and Hippo signaling pathways. Using mouse colonoids, we investigated the role of E-cadherin during intestinal homeostasis and regeneration using an IBD-like anoxia/cytokine model. We generated inducible E-cadherin shRNA knock-down mouse colonoids using lentiviral vectors. Acute injury was induced by subjecting colonoids to inflammatory cytokines, TNF-α and INF-γ, in anoxia, closely recapitulating the physiologic environment of IBD. Phenotypes and gene expression were analyzed by microscopy and real-time PCR. Our findings show that E-cadherin knockdown led to a more spherical and less differentiated morphology in colonoid cultures, accompanied by a significant increase in the mRNA expression of ISC marker Lgr5. The expression of reserve stem cell markers (Bmi1 and Hopx) remained unchanged, while markers for secretory progenitors were upregulated, and absorptive progenitor markers were downregulated. During regeneration after IBD-like anoxia/cytokine injury, characterized by increased area and bud number of colonoids, mRNA expression of reserve stem cell marker Hopx was considerably elevated with reduced E-cadherin expression. Notably, we observed increased expression of Wnt and YAP target genes and altered expression of Notch pathway genes in E-cadherin-deficient colonoids. These results suggest that E-cadherin regulates ISC development and intestinal regeneration through critical cellular pathways, including Wnt, Hippo, and Notch signaling pathways. Further studies on the mechanistic role of E-cadherin in ISC populations during regeneration, particularly in the context of IBD, may reveal novel therapeutic strategies aimed at enhancing mucosal healing and restoring intestinal homeostasis in IBD patients. Maryland Stem Cell Research Fund (Grant no. 2021-MSCRFL-5577) 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.