Angiogenic and pleiotropic effects of VEGF165 and HGF combined gene therapy in a rat model of myocardial infarction

Abstract

Since development of plasmid gene therapy for therapeutic angiogenesis by J. Isner this approach was an attractive option for ischemic diseases affecting large cohorts of patients. However, first placebo-controlled clinical trials showed its limited efficacy questioning further advance to practice. Thus, combined methods using delivery of several angiogenic factors got into spotlight as a way to improve outcomes. This study provides experimental proof of concept for a combined approach using simultaneous delivery of VEGF165 and HGF genes to alleviate consequences of myocardial infarction (MI). However, recent studies suggested that angiogenic growth factors have pleiotropic effects that may contribute to outcome so we expanded focus of our work to investigate potential mechanisms underlying action of VEGF165, HGF and their combination in MI. Briefly, Wistar rats underwent coronary artery ligation followed by injection of plasmid bearing VEGF165 or HGF or mixture of these. Histological assessment showed decreased size of post-MI fibrosis in both—VEGF165- or HGF-treated animals yet most prominent reduction of collagen deposition was observed in VEGF165+HGF group. Combined delivery group rats were the only to show significant increase of left ventricle (LV) wall thickness. We also found dilatation index improved in HGF or VEGF165+HGF treated animals. These effects were partially supported by our findings of c-kit+ cardiac stem cell number increase in all treated animals compared to negative control. Sporadic Ki-67+ mature cardiomyocytes were found in peri-infarct area throughout study groups with comparable effects of VEGF165, HGF and their combination. Assessment of vascular density in peri-infarct area showed efficacy of both–VEGF165 and HGF while combination of growth factors showed maximum increase of CD31+ capillary density. To our surprise arteriogenic response was limited in HGF-treated animals while VEGF165 showed potent positive influence on a-SMA+ blood vessel density. The latter hinted to evaluate infiltration of monocytes as they are known to modulate arteriogenic response in myocardium. We found that monocyte infiltration was driven by VEGF165 and reduced by HGF resulting in alleviation of VEGF-stimulated monocyte taxis after combined delivery of these 2 factors. Changes of monocyte infiltration were concordant with a-SMA+ arteriole density so we tested influence of VEGF165 or HGF on endothelial cells (EC) that mediate angiogenesis and inflammatory response. In a series of in vitro experiments we found that VEGF165 and HGF regulate production of inflammatory chemokines by human EC. In particular MCP-1 levels changed after treatment by recombinant VEGF, HGF or their combination and were concordant with NF-κB activation and monocyte infiltration in corresponding groups in vivo. We also found that both–VEGF165 and HGF upregulated IL-8 production by EC while their combination showed additive type of response reaching peak values. These changes were HIF-2 dependent and siRNA-mediated knockdown of HIF-2α abol...

Self‐assembling peptide modified with QHREDGS as a novel delivery system for mesenchymal stem cell transplantation after myocardial infarction

“…HGF could also activate endothelial cells migration, proliferation, and tube formation without increasing vascular permeability. Furthermore, it was shown that VEGF and HGF combined gene therapy alleviated consequences of MI (32). In the present study, treatment with culture supernatant from DSAP-MSCs markedly reduced the percentage of apoptotic cardiomyocytes and increased the level of p-Akt.…”

Effect of Co-culturing Fibroblasts in Human Skeletal Muscle Cell Sheet on Angiogenic Cytokine Balance and Angiogenesis

“…VEGF, HGF, and basic fibroblast growth factor (bFGF or FGF2), which are direct proangiogenic markers that promote angiogenesis (Fallah et al, 2019;Laddha and Kulkarni, 2019), are experimentally demonstrated to improve cardiac functions. Combined delivery of HGF and VEGF to infarcted myocardium showed an increase of left ventricle (LV) wall thickness and capillary density, reduce myocardial infarction size and improve dilatation index (Makarevich et al, 2018). Clinical trials have demonstrated enhancing myocardial perfusion leading to a better cardiac function and well-tolerated following therapy with VEGF, HGF, and FGF2 (Atluri and Woo, 2008).…”

HGF Treatment Promotes Cardiac Function and Cardiac Repair: Meta-analysis of Pig Models With Myocardial Infarction

“…A variety of vascular growth factors have been studied for gene therapy aiming at ameliorating cardiac function post myocardial injury, among which hepatocyte growth factor (HGF) and insulin-like growth factor (IGF-1) have attracted wide attention [6]. HGF, an important active factor for cell development and differentiation, could regulate the expression of cardiomyocyte speci c transcription factors and structural genes through its unique tyrosine kinase receptor c-Met [7], and then play functions such as promoting angiogenesis, regulating in ammation, inhibiting brosis, and activating tissue regeneration [8]. HGF and its c-Met receptor have been reported to alleviate chronic myocardial injury in acute MI, myocarditis, cardiomyopathy and other disease models [9].…”