Subepithelial Myofibroblasts Are Critical Regulators of Intestinal Epithelial Restoration

Abstract

Fibroblasts reside underneath most epithelial tissues. In the intestine, recent studies have shown that fibroblast migration contributes to tissue morphogenesis and wound healing. Yet, whether physical interactions between epithelial cells and fibroblasts contribute to epithelial movement remains elusive. Here, we show that subepithelial fibroblast alignment enhances directed and persistent migration of organoid-derived intestinal epithelia. Using a reconstituted epithelial-stromal gap-closure model, we demonstrate that direct contact with fibroblasts improves gap closure by promoting cell alignment, sustaining tissue integrity, and synchronizing crypt-villus migration. Fibroblasts undergo long-range ordering to align perpendicularly to the epithelial front and deposit protein paths that act as guidance features to direct epithelial migration. In parallel, epithelial cells acquire a wound-associated epithelial-like phenotype, but insufficient to explain the effects of fibroblast contact. Our findings uncover a dual role for intestinal fibroblasts in epithelial repair, coordinating both biochemical and physical cues to ensure efficient and cohesive migration.