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 4

Antifibrotic effect of Am80 administration on intestinal fibrosis in the DSS mouse model.

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Antifibrotic effect of Am80 administration on intestinal fibrosis in the...
(A) Schematic diagram showing the experimental setup. Oral administration of Am80 (3 mg/kg/day) or DMSO every day was initiated simultaneously with the third dose of DSS administration from days 43–63. (B) Colon tissue sections from WT mice after Am80 or DMSO administration for 9 weeks are analyzed for Meflin expression using ISH. Boxed areas are magnified in insets. Arrows denote Meflin+ cells. (C and D) Body weight changes over time and colon length at Day 63 are measured. (E–G) Colon tissue sections after Am80 administration are stained with H&E (E), IHC for α-SMA (F), and Sirius Red (G), followed by collagen layer thickness, quantification of Sirius Red+ areas, and fibrosis score. (F and G) (n = 6 and 7 for the control and Am80 groups, respectively). (H) Oral administration of Am80 (3 mg/kg/day) or DMSO every day to WT mice from weeks 10 to 13 after the 3 cycles of DSS administration. (I) Colon tissue sections are analyzed for Meflin expression using ISH. Boxed areas are magnified in insets. Arrows denote Meflin+ cells. (n = 4 and 5 for the control and Am80 groups, respectively). (J and K) Body weight changes over time and colon length at Day 92 are evaluated. (L–N) Representative images for H&E, IHC for α-SMA, and Sirius Red staining performed in colon tissue sections from WT mice after DSS and subsequent Am80 administrations (n = 7 and 5 for the control and Am80 groups, respectively). (B, F, G, I, M, and N) 5 HPFs per area were quantified for each mouse. Small gray dots indicate individual HPFs, and black triangles indicate mouse-level means used for statistical analysis. Scale bars: 40 μm. 2-tailed Student’s t tests. *P < 0.05; ** P < 0.01; *** P < 0.001; ****P < 0.0001.