Stiffness Restricts the Stemness of the Intestinal Stem Cells and Skews Their Differentiation Toward Goblet Cells

BACKGROUND & AIMS: Fibrosis and tissue stiffening are hallmarks of inflammatory bowel disease (IBD). We have hy-pothesized that the increased stiffness directly contributes to the dysregulation of the epithelial cell homeostasis in IBD. Here, we aim to determine the impact of tissue stiffening on the fate and function of the intestinal stem cells (ISCs). METHODS: We developed a long-term culture system consisting of 2.5-dimensional intestinal organoids grown on a hydrogel matrix with tunable stiffness. Single-cell RNA sequencing provided stiffness-regulated transcriptional signatures of the ISCs and their differentiated progeny. YAP-knockout and YAP-overexpression mice were used to manipulate YAP expres-sion. In addition, we analyzed colon samples from murine Exception & Big items.xlsx models and patients with IBD demonstrated cellular and molec-ular remodeling reminiscent of those observed in vitro. CON-CLUSIONS: Collectively, our findings highlight that matrix stiffness potently regulates the stemness of ISCs and their differentiation trajectory, supporting the hypothesis that fibrosis-induced gut stiffening plays a direct role in epithelial remodeling in IBD.

Automated summary

This study investigates the impact of tissue stiffening on the fate and function of intestinal stem cells (ISCs), and finds that matrix stiffness can regulate the stemness and differentiation trajectory of ISCs, supporting the direct role of fibrosis-induced gut stiffening in epithelial remodeling in inflammatory bowel disease (IBD).