Cell-Based HIF1α Gene Therapy Reduces Myocardial Scar and Enhances Angiopoietic Proteome, Transcriptomic and miRNA Expression in Experimental Chronic Left Ventricular Dysfunction

Abstract

Recent preclinical investigations and clinical trials with stem cells mostly studied bone-marrow-derived mononuclear cells (BM-MNCs), which so far failed to meet clinically significant functional study endpoints. BM-MNCs containing small proportions of stem cells provide little regenerative potential, while mesenchymal stem cells (MSCs) promise effective therapy via paracrine impact. Genetic engineering for rationally enhancing paracrine effects of implanted stem cells is an attractive option for further development of therapeutic cardiac repair strategies. Non-viral, efficient transfection methods promise improved clinical translation, longevity and a high level of gene delivery. Hypoxia-induced factor 1α is responsible for pro-angiogenic, anti-apoptotic and anti-remodeling mechanisms. Here we aimed to apply a cellular gene therapy model in chronic ischemic heart failure in pigs. A non-viral circular minicircle DNA vector (MiCi) was used for in vitro transfection of porcine MSCs (pMSC) with HIF1α (pMSC-MiCi-HIF-1α). pMSCs-MiCi-HIF-1α were injected endomyocardially into the border zone of an anterior myocardial infarction one month post-reperfused-infarct. Cell injection was guided via 3D-guided NOGA electro-magnetic catheter delivery system. pMSC-MiCi-HIF-1α delivery improved cardiac output and reduced myocardial scar size. Abundances of pro-angiogenic proteins were analyzed 12, 24 h and 1 month after the delivery of the regenerative substances. In a protein array, the significantly increased angiogenesis proteins were Activin A, Angiopoietin, Artemin, Endothelin-1, MCP-1; and remodeling factors ADAMTS1, FGFs, TGFb1, MMPs, and Serpins. In a qPCR analysis, increased levels of angiopeptin, CXCL12, HIF-1α and miR-132 were found 24 h after cell-based gene delivery, compared to those in untreated animals with infarction and in control animals. Expression of angiopeptin increased already 12 h after treatment, and miR-1 expression was reduced at that time point. In total, pMSC overexpressing HIF-1α showed beneficial effects for treatment of ischemic injury, mediated by stimulation of angiogenesis.

Recent advances in pre-conditioned mesenchymal stem/stromal cell (MSCs) therapy in organ failure; a comprehensive review of preclinical studies

“…Also, Gas6 could enhance VEGF, bFGF, SDF, and IGF-1 secretion from MSCs [ 116 ]. Likewise, HIF-1-overexpressing MSCs were found to increase cardiac output and decrease the size of myocardial scars in HF in vivo models [ 117 ]. Further, HIF-1 overexpression significantly augmented the secretion of angiogenesis proteins like activin A, angiopoietin, artemin, endothelin-1, MCP-1, and remodeling factors ADAMTS1, FGFs, TGF-β, MMPs, and serpins in MSCs in vitro [ 117 ].…”

“…Likewise, HIF-1-overexpressing MSCs were found to increase cardiac output and decrease the size of myocardial scars in HF in vivo models [ 117 ]. Further, HIF-1 overexpression significantly augmented the secretion of angiogenesis proteins like activin A, angiopoietin, artemin, endothelin-1, MCP-1, and remodeling factors ADAMTS1, FGFs, TGF-β, MMPs, and serpins in MSCs in vitro [ 117 ]. Genetically modified MSCs to overexpress VEGF in hypoxic conditions also increased myocardial neovascularization in ischemic heart disease [ 118 ].…”