TWIST1+FAP+ fibroblasts in the pathogenesis of intestinal fibrosis in Crohn’s disease
Yao Zhang, … , Duowu Zou, Bing Su
Yao Zhang, … , Duowu Zou, Bing Su
Published July 18, 2024
Citation Information: J Clin Invest. 2024;134(18):e179472. https://doi.org/10.1172/JCI179472.
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Research Article Gastroenterology

TWIST1+FAP+ fibroblasts in the pathogenesis of intestinal fibrosis in Crohn’s disease

Abstract

Intestinal fibrosis, a severe complication of Crohn’s disease (CD), is characterized by excessive extracellular matrix (ECM) deposition and induces intestinal strictures, but there are no effective antifibrosis drugs available for clinical application. We performed single-cell RNA sequencing (scRNA-Seq) of fibrotic and nonfibrotic ileal tissues from patients with CD with intestinal obstruction. Analysis revealed mesenchymal stromal cells (MSCs) as the major producers of ECM and the increased infiltration of its subset FAP+ fibroblasts in fibrotic sites, which was confirmed by immunofluorescence and flow cytometry. Single-cell transcriptomic profiling of chronic dextran sulfate sodium salt murine colitis model revealed that CD81+Pi16– fibroblasts exhibited transcriptomic and functional similarities to human FAP+ fibroblasts. Consistently, FAP+ fibroblasts were identified as the key subtype with the highest level of ECM production in fibrotic intestines. Furthermore, specific knockout or pharmacological inhibition of TWIST1, which was highly expressed by FAP+ fibroblasts, could significantly ameliorate fibrosis in mice. In addition, TWIST1 expression was induced by CXCL9+ macrophages enriched in fibrotic tissues via IL-1β and TGF-β signal. These findings suggest the inhibition of TWIST1 as a promising strategy for CD fibrosis treatment.

Authors

Yao Zhang, Jiaxin Wang, Hongxiang Sun, Zhenzhen Xun, Zirui He, Yizhou Zhao, Jingjing Qi, Sishen Sun, Qidi Yang, Yubei Gu, Ling Zhang, Chunhua Zhou, Youqiong Ye, Ningbo Wu, Duowu Zou, Bing Su

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Figure 7

Targeting TWIST1 inhibits fibroblast activation and attenuates experimental intestinal fibrosis.

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Targeting TWIST1 inhibits fibroblast activation and attenuates experimen...
(A) Western blotting images showing the expression of ECM-related genes in primary human intestinal fibroblasts with or without TGF-β (5 ng/mL, 48 hours) and harmine administration (5 μM or 10 μM, 48 hours). (B) Schematic diagram for the in vivo experiments (5 mice per group). (C) Masson’s trichrome staining showing collagen deposition in mouse colons across the 4 indicated groups. Scale bar: 100 μm. (D) Bar plots showing histologic scores of mouse colons across the 4 indicated groups. Data represent the mean ± SD. Statistical differences were determined by the 1-way ANOVA with Bonferroni’s correction. (E) Representative IF staining of mouse colons across the 4 indicated groups (original magnification, ×20). DAPI (blue), COL1A1 (red), and vimentin (green) in merged channels are shown. Scale bar: 50 μm. (F) Quantitative analysis (integrated fluorescence intensity) of COL1A1 in IF staining of mouse colons. Data represent the mean ± SD. Statistical differences were determined by 1-way ANOVA with Bonferroni’s correction. (G and H) Representative plots (G) and quantitative analysis (H) of Western blotting images showing the expression of fibronectin and TWIST1 in mouse colons across the 4 indicated groups. Data represent the mean ± SD. Statistical differences were determined by 1-way ANOVA with Bonferroni’s correction.