LAUSR

Myocardial infarction (MI) and hypertension lead to myocardial injury, which induces negative remodelling and cardiac fibrosis. Cardiac fibrosis, which involves inflammatory cell infiltration and myofibroblast activation, causes worldwide mortality and morbidity. In response to MI or hypertension induced by prolonged angiotensin II (AngII) exposure, activated myofibroblasts produce extracellular matrix proteins. However, if unchecked, excessive collagen deposition occurs leading to myocardial stiffening, heart failure and arrhythmias. The underlying mechanisms leading to pathological collagen deposition are not fully elucidated. There is debate regarding the involvement of the Wnt signalling pathway and its product Wnt Inducible Signalling pathway protein-1 (WISP-1) in cardiac fibrosis. Therefore, this project aimed to investigate the interaction of AngII and the Wnt/β-catenin signalling pathway in cardiac fibrosis. The effect of AngII (100nM) on collagen levels in human cardiac fibroblasts was investigated in vitro (data expressed as fold change from control ± SEM). In vivo experiments (n=6–8) determined the involvement of the Wnt/b-catenin pathway, specifically WISP-1, in response to AngII infusion (500ng/kg/min) for 4 weeks (Apolipoprotein E−/−/WISP-1+/+ vs. ApolipoproteinE−/−/WISP-1−/− mice on a high-fat diet, data expressed as mean positive pixel % ± SEM). AngII significantly increased collagen type 1 protein levels produced by human cardiac fibroblasts (2.94±0.75 vs 1±0, p<0.05). Inhibition of Wnt/b-catenin signalling with 25nM iCRT14 significantly suppressed AngII-induced collagen levels (0.46±0.07 vs. 1±0, p<0.05). As expected, AngII infusion significantly induced hypertension in all mice. Immunohistochemistry demonstrated type 1 collagen was markedly higher in AngII mice than control mice (1.07±0.27 vs. 0.29±0.06, p<0.05). However, in the absence of WISP-1, AngII did not enhance collagen type 1 levels. Further immunohistochemical analysis of murine hearts demonstrated that AngII infusion caused significant alterations in the Wnt/β-catenin signalling markers AXIN-2 (35±3.9 vs. 10.7±2.6 p<0.05) and PPAR-d (92±1.4 vs. 17.3±4.3 p<0.05). This effect was reduced by WISP-1 deletion. Furthermore, AngII-infusion disrupted N-cadherin junctions (0.55±0.08 vs. 0.29±0.02 p<0.05) suggesting modulation of cell-to-cell contacts and enhanced β-catenin signalling. This study indicates that AngII enhances cardiac fibrosis via modulation of the Wnt signalling pathway, in part via WISP-1. Further delineation of this interaction may lead to the use of Wnt/β-catenin or WISP-1 inhibitors to suppress myocardial injury induced cardiac fibrosis in post-MI or hypertensive patients. Elizabeth Blackwell Institute, British Heart Foundation