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

Shijie He, Peng Lei, Wenying Kang, Priscilla Cheung, Tao Xu, Miyeko Mana, Chan Young Park, Hongyan Wang, Shinya Imada, Jacquelyn O. Russell, Jianxun Wang, Ruizhi Wang, Ziheng Zhou, Kashish Chetal, Eric Stas, Vidisha Mohad, Peter Bruun-Rasmussen, Ruslan I. Sadreyev, Richard A. Hodin, Yanhang ZhangDavid T. Breault, Fernando D. Camargo, Ömer H. Yilmaz, Jeffrey J. Fredberg, Nima Saeidi

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Background & Aims: Fibrosis and tissue stiffening are hallmarks of inflammatory bowel disease (IBD). We have hypothesized 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 expression. In addition, we analyzed colon samples from murine colitis models and human IBD samples to assess the impact of stiffness on ISCs in vivo. Results: We demonstrated that increasing the stiffness potently reduced the population of LGR5+ ISCs and KI-67+–proliferating cells. Conversely, cells expressing the stem cell marker, olfactomedin-4, became dominant in the crypt-like compartments and pervaded the villus-like regions. Concomitantly, stiffening prompted the ISCs to preferentially differentiate toward goblet cells. Mechanistically, stiffening increased the expression of cytosolic YAP, driving the extension of olfactomedin-4+ cells into the villus-like regions, while it induced the nuclear translocation of YAP, leading to preferential differentiation of ISCs toward goblet cells. Furthermore, analysis of colon samples from murine colitis models and patients with IBD demonstrated cellular and molecular remodeling reminiscent of those observed in vitro. Conclusions: 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.

Original languageEnglish (US)
Pages (from-to)1137-1151.e15
JournalGastroenterology
Volume164
Issue number7
DOIs
StatePublished - Jun 2023
Externally publishedYes

Keywords

  • Fibrosis
  • IBD
  • Intestinal Organoids
  • Intestinal Stem Cells
  • Stiffening

ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology

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