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 7

Am80 administration reverses fibrotic response of fibroblasts and tissue pieces isolated from patients with CD.

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Am80 administration reverses fibrotic response of fibroblasts and tissue...
(A) Intestinal fibroblasts and ileal tissue pieces are isolated from the intestinal nonstenotic and stenotic areas surgically resected from patients with CD, followed by culture on plastic. These samples are treated with Am80 (50, 100, 300 μg/mL) or DMSO for 48 or 72 hours, followed by the indicated experiments. (B) Primary cultured intestinal fibroblasts from the nonstenotic (left) and stenotic (right) areas are fixed and stained for α-SMA using IF. Arrows denote fibroblasts that are strongly positive for α-SMA. Scale bars: 20 μm. (C) qPCR analysis for Meflin mRNA expression in the primary cultured intestinal fibroblasts (n = 4 and 5 for the nonstenotic and stenotic groups, respectively). (D) Intestinal fibroblasts cultured in the absence (left) or presence (right) of Am80 are fixed and subjected to IF for α-SMA. Arrows denote fibroblasts that are strongly positive for α-SMA. Scale bars: 20 μm. (E) qPCR analysis for the expression of the indicated genes in the intestinal fibroblasts cultured in the absence (left) or presence (right) of Am80 (n = 5 for each group). (F and G) Ileal tissue pieces obtained from the stenotic regions of intestine are incubated with different concentrations of Am80 (50 and 300 μg/mL) or DSMO for 24 hours, followed by the measurement of TGF-β (F) and IL-6 (G) in the culture supernatants using enzyme-linked immunosorbent assays (n = 7 to 12 for each group). (C, E, F, G) Statistical significance was determined using 2-tailed Student’s t tests. *P < 0.05; **P < 0.01; ****P < 0.0001.