Endoglin negatively regulates transforming growth factor β1-induced profibrotic responses in intestinal fibroblasts

J. P. Burke, R. W.G. Watson, J. J. Mulsow, N. G. Docherty, J. C. Coffey, P. R. O'Connell

Research output: Contribution to journalArticlepeer-review

Abstract

Background: Fibroblasts isolated from strictures in Crohn's disease (CD) exhibit reduced responsiveness to stimulation with transforming growth fector (TGF) β1. TGF-β1, acting through the smad pathway, is critical to fibroblast-mediated intestinal fibrosis. The membrane glycoprotein, endoglin, is a negative regulator of TGF-β. Methods: Intestinal fibroblasts were cultured from seromuscular biopsies of patients undergoing intestinal resection for CD strictures or from control patients. Endoglin expression was assessed using confocal microscopy, flow cytometry and western blot. The effect of small interfering (si) RNAmediated knockdown and plasmid-mediated overexpression of endoglin on fibroblast responsiveness to TGF-β1 was assessed by examining smad phosphorylation, smad binding element (SBE) promoter activity, connective tissue growth factor (CTGF) expression and ability to contract collagen. Results: Crohn's stricture fibroblasts expressed increased constitutive cell-surface and whole-cell endoglin relative to control cells. Endoglin co-localized with filamentous actin. Fibroblasts treated with siRNA directed against endoglin exhibited enhanced TGF-β1-mediated smad-3 phosphorylation, and collagen contraction. Cells transfected with an endoglin plasmid did not respond to TGF-β1 by exhibiting SBE promoter activity or producing CTGF. Conclusion: Fibroblasts from strictures in CD express increased constitutive endoglin. Endoglin is a negative regulator of TGF-β1 signalling in the intestinal fibroblast, modulating smad-3 phosphorylation, SBE promoter activity, CTGF production and collagen contraction.

Original languageEnglish
Pages (from-to)892-901
Number of pages10
JournalBritish Journal of Surgery
Volume97
Issue number6
DOIs
Publication statusPublished - Jun 2010
Externally publishedYes

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