Potential therapeutic efficacy via avB3-integrin dependent interaction of developmental endothelial locus-1 (Del-1) after established angiotensin-II dependent hypertension

Abstract

Abstract Introduction Cardiovascular diseases (CVDs) contribute to about 31% of global death. Hypertension is a major risk factor and also increases the risk of CVDs. An important regulator of systemic blood pressure in the body, the Renin Angiotensin Aldosterone system becomes overactivated during hypertension releasing a critical product, angiotensin-II (ANG-II), a known vasoconstrictor, activating the immune system leading to inflammation and as such mediating the effects of hypertension and subsequent end-organ damage due to the critical role of the innate and adaptive cells in producing pro-inflammatory cytokines such interleukin-17. Del-1, an extracellular matrix protein expressed mostly by endothelial cells is a known potent inhibitor of leukocyte recruitment. Aim We tested the hypothesis that, Del-1 via αvβ3-integrin dependent interaction protects against ANG-II induced cardiovascular remodeling after established hypertension. Methods Wild type (WT) mice were infused with 2 mg/kg/day of ANG-II for 14 days. Soluble Del-1 (Del-1-Fc) and a point mutant unable to bind to αvβ3-integrin (Del-1-RGE-Fc) was injected (50 μg/per injection) daily after 6 days of ANG-II infusion. SBP was measured using tail-cuff. Endothelial function was assessed using Mulvany myograph and quantification of inflammatory cells and IL-17 by flow cytometry. Histological staining for fibrosis, myocardial/medial hypertrophy and elastin degradation was used to evaluate both aortic and cardiac remodeling. Results After 6 days, mice infused with ANG-II showed significantly higher SBP of 140 mmHg as compared to control mice (110 mmHg). However, further progression of SBP after 6 days was blunted in infused mice treated with soluble Del-1 (ANGII+Del-1-Fc) as compared to similar increase in SBP in both infused mice treated with the point mutant (ANGII+Del-1-RGE-Fc) and non-treated infused mice (ANGII+Fc) leading to a difference of ∼20 mmHg. This effect was due to, the well protected endothelium dependent relaxation and reduced wall tension of the aorta in ANGII+Del-1-Fc mice. Further infiltration of CD45+ leukocytes, CD45+/IL-17A+ and TCRβ+T cells was significantly reduced in the aorta and heart of ANGII+Del-1-Fc mice as compared to both ANGII+Del-1-RGE-Fc and ANGII+Fc mice. However, the aorta and heart of ANGII+Del-1-Fc mice showed increased levels of anti-inflammatory regulatory T-cells (CD25+FOXP3+Treg). Also, aortic medial thickness, adventitial collagen and elastin degradation was decreased significantly in ANGII+Del-1-Fc mice. Additionally, in comparison to both ANGII+Del-1-RGE-Fc and ANGII+Fc mice, the hearts of ANGII+Del-1-Fc mice had significantly less interstitial and pericoronary collagen, smaller cardiomyocyte area and subsequently higher lumen-to-wall ratio. Conclusion Our study reveals the cardiovascular protective effects of Del-1 mediated via its association with αvβ3-integrin after established hypertension which could serve as potential therapeutic target. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): German Research Foundation (DFG)National Institutes of Health (NIH)