Direct signaling of TL1A-DR3 on fibroblasts induces intestinal fibrosis in vivo

Tumor necrosis factor-like cytokine 1A (TL1A, TNFSF15) is implicated in inflammatory bowel disease, modulating the location and severity of inflammation and fibrosis. TL1A expression is increased in inflamed mucosa and associated with fibrostenosing Crohn’s disease. Tl1a-overexpression in mice causes spontaneous ileitis, and exacerbates induced proximal colitis and fibrosis. Intestinal fibroblasts express Death-receptor 3 (DR3; the only know receptor for TL1A) and stimulation with TL1A induces activation in vitro. However, the contribution of direct TL1A-DR3 activation on fibroblasts to fibrosis in vivo remains unknown. TL1A overexpressing naïve T cells were transferred into Rag(−/−) , Rag(−/−) mice lacking DR3 in all cell types (Rag(−/−)Dr3(−/−)), or Rag(−/−) mice lacking DR3 only on fibroblasts (Rag(−/−)Dr3(∆Col1a2)) to induce colitis and fibrosis, assessed by clinical disease activity index, intestinal inflammation, and collagen deposition. Rag(−/−) mice developed overt colitis with intestinal fibrostenosis. In contrast, Rag(−/−)Dr3(−/−) demonstrated decreased inflammation and fibrosis. Despite similar clinical disease and inflammation as Rag(−/−), Rag(−/−)Dr3(∆Col1a2) exhibited reduced intestinal fibrosis and attenuated fibroblast activation and migration. RNA-Sequencing of TL1A-stimulated fibroblasts identified Rho signal transduction as a major pathway activated by TL1A and inhibition of this pathway modulated TL1A-mediated fibroblast functions. Thus, direct TL1A signaling on fibroblasts promotes intestinal fibrosis in vivo. These results provide novel insight into profibrotic pathways mediated by TL1A paralleling its pro-inflammatory effects.