Treatment of intestinal fibrosis in experimental inflammatory bowel disease by the pleiotropic actions of a local Rho kinase inhibitor

• Published in: Gastroenterology (New York, N.Y. 1943) Vol. 153; no. 4; pp. 1054 - 1067
• Main Authors: Holvoet, Tom, Devriese, Sarah, Castermans, Karolien, Boland, Sandro, Leysen, Dirk, Vandewynckel, Yves-Paul, Devisscher, Lindsey, Van den Bossche, Lien, Van Welden, Sophie, Dullaers, Melissa, Vandenbroucke, Roosmarijn E, De Rycke, Riet, Geboes, Karel, Bourin, Arnaud, Defert, Olivier, Hindryckx, Pieter, De Vos, Martine, Laukens, Debby
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.2017
• Subjects: Adoptive Transfer; Animals; Autophagy - drug effects; Case-Control Studies; Colitis; Collagen - metabolism; Dextran Sulfate; Disease Models, Animal; Enzyme Activation; Epithelial-to-Mesenchymal Transition; Fibrosis; Gastroenterology and Hepatology; Humans; Ileum - drug effects; Ileum - enzymology; Ileum - immunology; Ileum - pathology; Inflammatory Bowel Diseases - chemically induced; Inflammatory Bowel Diseases - enzymology; Inflammatory Bowel Diseases - pathology; Inflammatory Bowel Diseases - prevention & control; Interleukin-6 - metabolism; Intestinal Obstruction - chemically induced; Intestinal Obstruction - enzymology; Intestinal Obstruction - pathology; Intestinal Obstruction - prevention & control; Male; Matrix Metalloproteinases - metabolism; Mesenchymal Cells; Mice, Inbred C57BL; Myofibroblasts - drug effects; Myofibroblasts - enzymology; Myofibroblasts - immunology; Myofibroblasts - pathology; p38 Mitogen-Activated Protein Kinases - metabolism; Protein Kinase Inhibitors - pharmacology; rho-Associated Kinases - antagonists & inhibitors; rho-Associated Kinases - metabolism; Signal Transduction - drug effects; Stenosis; T-Lymphocytes - immunology; T-Lymphocytes - transplantation; Time Factors; Tissue Culture Techniques; colitis; 2, 4, 6-trinitrobenzenesulfonic acid; MLC; autophagy-related 16 like 1; alpha smooth muscle actin; MTT; MPO; TNF; UC; CTGF; myosin light chain; ulcerative colitis; human intestinal fibroblasts; IL; DSS; dextran sulfate sodium; lactate dehydrogenase; EMT; inflammatory bowel disease; epithelial-to-mesenchymal transition; connective tissue growth factor; MMP; severe combined immunodeficient; TGF; lipopolysaccharide; ROCK; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; IBD; stenosis; SCID; LPS; IFN; LDH; MCP1; myocardin-related transcription factor; interleukin; Crohn’s disease; CD; HIF; myeloperoxidase; interferon; TNBS; ECM; Rho-associated coiled-coil-containing protein kinase; MRTF; tumor necrosis factor; ATG16L1; monocyte chemo-attractant protein 1; LC; αSMA; microtubule-associated protein light chain; matrix metalloproteinase; transforming growth factor; mesenchymal cells; extracellular matrix; Stenosis; IP; qRT-PCR; Epithelial-to-Mesenchymal Transition; Colitis; Mesenchymal Cells
• ISSN: 0016-5085; 1528-0012
• DOI: 10.1053/j.gastro.2017.06.013

Abstract Background Intestinal fibrosis resulting in (sub)obstruction is a common complication of Crohn’s disease (CD). Rho kinases (ROCKs) play multiple roles in TGFβ1-induced myofibroblast activation that could be therapeutic targets. Because systemic ROCK inhibition causes cardiovascular side-effects, we evaluated the effects of a locally acting ROCK inhibitor (AMA0825) on intestinal fibrosis. Methods Fibrosis was assessed in mouse models using dextran sulfate sodium and adoptive T cell transfer. The in vitro and ex vivo effects of AMA0825 were studied in different cell types and in CD biopsy cultures. Results ROCK is expressed in fibroblastic, epithelial, endothelial and muscle cells of the human intestinal tract and is activated in inflamed and fibrotic tissue. Prophylactic treatment with AMA0825 inhibited myofibroblast accumulation, expression of pro-fibrotic factors and accumulation of fibrotic tissue without affecting clinical disease activity and histological inflammation in two models of fibrosis. ROCK inhibition reversed established fibrosis in a chronic DSS model and impeded ex vivo pro-fibrotic protein secretion from stenotic CD biopsies. AMA0825 reduced TGFβ1-induced activation of myocardin-related transcription factor (MRTF) and p38 mitogen-activated protein kinase (MAPK), downregulating matrix metalloproteinases, collagen and IL6 secretion from fibroblasts. In these cells, ROCK inhibition potentiated autophagy, which was required for the observed reduction in collagen and IL6 production. AMA0825 did not affect pro-inflammatory cytokine secretion from other ROCK-positive cell types, corroborating the selective in vivo effect on fibrosis. Conclusions Local ROCK inhibition prevents and reverses intestinal fibrosis by diminishing MRTF and p38 MAPK activation and increasing autophagy in fibroblasts. Overall, our results show that local ROCK inhibition is promising for counteracting fibrosis as an add-on therapy for CD.