Meflin confers antifibrotic properties to intestinal fibroblasts in inflammatory bowel disease
Jingxi Mu, Keiko Maeda, Tadashi Iida, Shinji Mii, Nobutoshi Esaki, Yukihiro Shiraki, Yasuyuki Mizutani, Masanao Nakamura, Takeshi Yamamura, Tsunaki Sawada, Eri Ishikawa, Kentaro Murate, Takashi Hirose, Kazuhiro Furukawa, Akina Oishi, Haruhiko Suzuki, Takayoshi Kishida, Goro Nakayama, Mitsuhiro Fujishiro, Hiroki Kawashima, Atsushi Enomoto
Jingxi Mu, Keiko Maeda, Tadashi Iida, Shinji Mii, Nobutoshi Esaki, Yukihiro Shiraki, Yasuyuki Mizutani, Masanao Nakamura, Takeshi Yamamura, Tsunaki Sawada, Eri Ishikawa, Kentaro Murate, Takashi Hirose, Kazuhiro Furukawa, Akina Oishi, Haruhiko Suzuki, Takayoshi Kishida, Goro Nakayama, Mitsuhiro Fujishiro, Hiroki Kawashima, Atsushi Enomoto
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Research Article Gastroenterology Inflammation

Meflin confers antifibrotic properties to intestinal fibroblasts in inflammatory bowel disease

Abstract

Dysfunctional intestinal fibrosis is an irreversible complication of Crohn’s disease (CD). The complex heterogeneity of intestinal mesenchymal cells makes it difficult to understand the pathogenesis of intestinal fibrosis. Previously, we identified Meflin as a marker of fibroblast subsets. This study aimed to explore the role of Meflin-positive fibroblasts in intestinal fibrogenesis and investigate the potential of pharmacological control of Meflin expression as a treatment for patients with CD. Our results indicated that Meflin expression was upregulated in fibroblasts at the early stage of fibrosis but was downregulated in established fibrosis in both patients with CD and 2 different mouse models, which are the chronic dextran sodium sulfate (DSS) model and an IL-10–deficient model that spontaneously develops intestinal inflammation. Meflin-deficient mice exacerbated intestinal fibrosis with dysregulated expression of noncanonical Wnt ligand WNT5A and its receptor ROR2. Pharmacologically induced Meflin expression through the administration of a synthetic retinoid reversed intestinal fibrosis in the DSS model and suppressed profibrotic protein secretion in fibroblasts isolated from patients with CD. Our findings indicate that Meflin-positive fibroblasts represent a functional subpopulation that suppresses intestinal fibrosis. Augmentation of Meflin expression shows antifibrotic effects and holds promise as a therapeutic approach for intestinal fibrosis in patients with CD.

Authors

Jingxi Mu, Keiko Maeda, Tadashi Iida, Shinji Mii, Nobutoshi Esaki, Yukihiro Shiraki, Yasuyuki Mizutani, Masanao Nakamura, Takeshi Yamamura, Tsunaki Sawada, Eri Ishikawa, Kentaro Murate, Takashi Hirose, Kazuhiro Furukawa, Akina Oishi, Haruhiko Suzuki, Takayoshi Kishida, Goro Nakayama, Mitsuhiro Fujishiro, Hiroki Kawashima, Atsushi Enomoto

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Figure 8

Meflin may inhibit intestinal fibrosis by suppressing the expression of WNT5A and its receptor ROR2.

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Meflin may inhibit intestinal fibrosis by suppressing the expression of ...
(A) Colonic fibroblasts isolated from WT mice after 9 weeks of DSS administration were transduced with control or mouse Meflin (mMeflin) cDNA using a lentiviral expression system, followed by qPCR analysis (n = 3 per group). CTL, control. 2-tailed Student’s t tests. (B) qPCR analysis for the expression of Fn1, Col3a1, and Col6a1 in the indicated fibroblasts (n = 3–6 per group). (C) Colonic fibroblasts isolated from WT and Meflin-KO mice after 9 weeks of DSS administration were cultured with or without recombinant BMP7 (rBMP7, 100 ng/mL) for 1 hour, followed by Western blot analysis for WNT5A/B and β-actin. The data from 2 independent experimental batches are shown. kDa, kilodaltons. (D) Colonic fibroblasts isolated from nonstricture and stricture areas of patients with CD were treated with Am80 (1 μM) for 48 hours, followed by Western blot analysis for Meflin, WNT5A/B, and GAPDH. (E) qPCR analysis for Ror2 mRNA expression in colonic fibroblasts isolated from WT and Meflin-KO mice. Restoring Meflin expression (mMeflin) in Meflin-KO fibroblasts significantly suppressed Ror2 expression (n = 6 per group). (F) Colonic fibroblasts isolated from WT and Meflin-KO mice were treated with rBMP7 (20 ng/ml) for 48 hours, followed by qPCR analysis for Ror2 mRNA expression (n = 9 per group). (G and H) Colonic fibroblasts were transfected with control siRNA (siControl) or siRor2, followed by qPCR analysis for Ror2 (G) in Col1a1 (H) mRNA levels (n = 3 per group). (I) Summary of the findings obtained by colonic fibroblasts from WT and Meflin-KO mice and Ror2 knockdown. (B, E, F) Each dot represents an individual sample. Scale bars: 40 μm. 1-way ANOVA unless otherwise indicated. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.