title: IL-1β stimulation of CCD-18co myofibroblasts enhances repair of epithelial monolayers through Wnt-5a.
abstract: Subepithelial myofibroblasts are involved in the initiation and coordination of intestinal epithelial repair, but the molecular signaling pathways are largely unknown. The cellular adaptations that occur during repair range from dedifferentiation and migration to proliferation and redifferentiation, in a way that is strongly reminiscent of normal crypt-to-villus epithelial maturation. We therefore hypothesized that Wnt/β-catenin signaling may have a pivotal role in intestinal epithelial wound repair. We used the established scratch wound method in Caco-2 cells and in nontransformed NCM460 cells to monitor the effects of IL-1β-stimulated colonic myofibroblasts (CCD-18co) on intestinal epithelial repair, with immunoblotting and immunodepletion to examine the conditioned media. Conditioned media from IL-1β-stimulated, but not -untreated, myofibroblasts increased Caco-2 wound closure twofold over 24 h. IL-1β-stimulated myofibroblasts downregulated the differentiation marker sucrase-isomaltase in the Caco-2 cells, whereas the proliferation marker c-myc was upregulated. Array expression profiling identified Wnt-5a as the Wnt-related gene that was most upregulated (28-fold) by IL-1β stimulation of CCDs. Recombinant Wnt-5a enhanced proliferation of Caco-2 and NCM460 cells. In scratch assays, it increased migration of the leading edge in both cell lines. Wnt-5a immunodepletion of the IL-1β-CCD conditioned media abrogated the ability to enhance the repair. Wnt-5a often acts through a noncanonical signal transduction pathway. Further experiments supported this pathway in epithelial wound healing: IL-1β-CCD-mediated repair was not affected by the addition of the canonical Wnt antagonist Dickkopf-1. Furthermore, media from stimulated myofibroblasts (but not Wnt-5a-depleted media) increased c-jun in Caco-2 cell nuclear extracts. Myofibroblast-mediated noncanonical Wnt-5a signaling is therefore important in the dedifferentiation and migration stages of epithelial wound repair.
authors.affiliation: Centre for Digestive Diseases, Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary, University of London, 4 Newark St., London E1 2AT, UK.
authors.forename: Meera
authors.initials: M
authors.lastname: Raymond
authors.forename: Tania
authors.initials: T
authors.lastname: Marchbank
authors.forename: Mary P
authors.initials: MP
authors.lastname: Moyer
authors.forename: Raymond J
authors.initials: RJ
authors.lastname: Playford
authors.forename: Ian R
authors.initials: IR
authors.lastname: Sanderson
authors.forename: Laurens
authors.initials: L
authors.lastname: Kruidenier
mesh_terms: D018938:Caco-2 Cells; D054337:Cell Dedifferentiation; D002460:Cell Line; D002465:Cell Movement; D017077:Culture Media, Conditioned / Q000494:pharmacology; D015536:Down-Regulation; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D058628:Myofibroblasts / Q000187:drug effects* / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects; D015854:Up-Regulation; D051153:Wnt Proteins / Q000096:biosynthesis / Q000502:physiology*; D000071818:Wnt-5a Protein; D014945:Wound Healing / Q000187:drug effects* / Q000502:physiology; D051176:beta Catenin / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D053583:Interleukin-1beta; D011518:Proto-Oncogene Proteins; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D051176:beta Catenin