Novel Designed Proteolytically Resistant VEGF-B186R127S Promotes Angiogenesis in Mouse Heart by Recruiting Endothelial Progenitor Cells

Abstract

Background: Previous studies have indicated that vascular endothelial growth factor B186 (VEGF-B186) supports coronary vascular growth in normal and ischemic myocardium. However, previous studies also indicated that induction of ventricular arrhythmias is a severe side effect preventing the use of VEGF-B186 in cardiac gene therapy, possibly mediated by binding to neuropilin 1 (NRP1). We have designed a novel VEGF-B186 variant, VEGF-B186R127S, which is resistant to proteolytic processing and unable to bind to NRP1. Here, we studied its effects on mouse heart to explore the mechanism of VEGF-B186-induced vascular growth along with its effects on cardiac performance.Methods: Following the characterization of VEGF-B186R127S, we performed ultrasound-guided adenoviral VEGF-B186R127S gene transfers into the murine heart. Vascular growth and heart functions were analyzed using immunohistochemistry, RT-PCR, electrocardiogram and ultrasound examinations. Endothelial progenitor cells (EPCs) were isolated from the circulating blood and characterized. Also, in vitro experiments were carried out in cardiac endothelial cells with adenoviral vectors.Results: The proteolytically resistant VEGF-B186R127S significantly induced vascular growth in mouse heart. Interestingly, VEGF-B186R127S gene transfer increased the number of circulating EPCs that secreted VEGF-A. Other proangiogenic factors were also present in plasma and heart tissue after the VEGF-B186R127S gene transfer. Importantly, VEGF-B186R127S gene transfer did not cause any side effects, such as arrhythmias.Conclusion: VEGF-B186R127S induces vascular growth in mouse heart by recruiting EPCs. VEGF-B186R127S is a novel therapeutic agent for cardiac therapeutic angiogenesis to rescue myocardial tissue after an ischemic insult.

Growth factor for therapeutic angiogenesis in ischemic heart disease: A meta-analysis of randomized controlled trials

“…Even though antiplatelet agent and statin are cornerstones for treating IHD, however, previous clinical studies showed that statin and aspirin was effective in decreasing VEGF levels and have no effects on promoting angiogenesis (Dworacka et al, 2014;Cheng et al, 2015). Recently, animal studies showed that VEFG for therapeutic angiogenesis could promotes collateral circulation in mouse heart by recruiting endothelial progenitor cells, and subsequently rescue myocardial tissue after an ischemic insult (Mallick et al, 2022). Transforming growth factor beta (TGF-β1) induces pro-reparative phenotypic changes in epicardial cells in mice after myocardial infarction (Dergilev et al, 2021).…”

Therapeutic Potential of VEGF-B in Coronary Heart Disease and Heart Failure: Dream or Vision?

“…The functions of VEGF-B are mediated by its binding to a tyrosine kinase receptor, named as vascular endothelial growth factor receptor 1 (VEGF-R1) and common co-receptors neuropilin 1 (NRP-1) or NRP-2 [ 27 , 28 , 29 ]. It needs to be pointed out that VEGF-B186 binds to VEGF-R1 before and after proteolytic cleavage but requires proteolytic cleavage to bind to NRP-1 or NRP-2 [ 28 , 30 ].…”

“…The functions of VEGF-B are mediated by its binding to a tyrosine kinase receptor, named as vascular endothelial growth factor receptor 1 (VEGF-R1) and common co-receptors neuropilin 1 (NRP-1) or NRP-2 [ 27 , 28 , 29 ]. It needs to be pointed out that VEGF-B186 binds to VEGF-R1 before and after proteolytic cleavage but requires proteolytic cleavage to bind to NRP-1 or NRP-2 [ 28 , 30 ]. VEGF-A does not seem to significantly activate VEGF-R1 downstream signaling compared with VEGF-R2 mediated downstream signaling, while VEGF-B binding to VEGF-R1 leads to the activation of AKT, PI3K, ERK, and MAPK [ 31 ].…”

“…In connection with coronary vessels, VEGF-B promotes angiogenesis in the subendocardial region and rescues the myocardium after myocardial infarction [ 52 ]. Our studies have shown that adenoviral vector-based VEGF-B186 gene transfer induced significant angiogenesis in the myocardium and improved myocardial perfusion without increasing a coronary steal effect [ 28 , 43 , 45 , 55 ]. Thus, VEGF-B186 seems highly beneficial for the treatment of CHD [ 43 , 45 ].…”

“…To resolve these questions, we have designed several VEGF-B186 isoforms, based on its structure, proteolytic processing, and known receptor binding properties [ 45 ]. The studies have found that only the unprocessed full-length form of VEGF-B186 induces angiogenesis in the heart and NRP binding is dispensable for the VEGF-B induced microvascular growth in the heart muscle [ 28 , 45 ]. Interestingly, despite having VEGF-R1 binding ability, the N-terminal forms of VEGF-B186 (e.g., VEGF-B127 and VEGF-B109) did not show any angiogenic activity in the heart [ 45 ].…”