Extracellular vesicles from human embryonic stem cell-derived cardiovascular progenitor cells promote cardiac infarct healing through reducing cardiomyocyte death and promoting angiogenesis

Abstract

Human pluripotent stem cells (hPSCs)-derived cardiovascular progenitor cells (CVPCs) are a promising source for myocardial repair, while the mechanisms remain largely unknown. Extracellular vesicles (EVs) are known to mediate cell-cell communication, however, the efficacy and mechanisms of hPSC-CVPC-secreted EVs (hCVPC-EVs) in the infarct healing when given at the acute phase of myocardial infarction (MI) are unknown. Here, we report the cardioprotective effects of the EVs secreted from hESC-CVPCs under normoxic (EV-N) and hypoxic (EV-H) conditions in the infarcted heart and the long noncoding RNA (lncRNA)-related mechanisms. The hCVPC-EVs were confirmed by electron microscopy, nanoparticle tracking, and immunoblotting analysis. Injection of hCVPC-EVs into acutely infracted murine myocardium significantly improved cardiac function and reduced fibrosis at day 28 post MI, accompanied with the improved vascularization and cardiomyocyte survival at border zones. Consistently, hCVPC-EVs enhanced the tube formation and migration of human umbilical vein endothelial cells (HUVECs), improved the cell viability, and attenuated the lactate dehydrogenase release of neonatal rat cardiomyocytes (NRCMs) with oxygen glucose deprivation (OGD) injury. Moreover, the improvement of the EV-H in cardiomyocyte survival and tube formation of HUVECs was significantly better than these in the EV-N. RNA-seq analysis revealed a high abundance of the lncRNA MALAT1 in the EV-H. Its abundance was upregulated in the infarcted myocardium and cardiomyocytes treated with hCVPC-EVs. Overexpression of human MALAT1 improved the cell viability of NRCM with OGD injury, while knockdown of MALAT1 inhibited the hCVPC-EV-promoted tube formation of HUVECs. Furthermore, luciferase activity assay, RNA pull-down, and manipulation of miR-497 levels showed that MALAT1 improved NRCMs survival and HUVEC tube formation through targeting miR-497. These results reveal that hCVPC-EVs promote the infarct healing through improvement of cardiomyocyte survival and angiogenesis. The cardioprotective effects of hCVPC-EVs can be enhanced by hypoxia-conditioning of hCVPCs and are partially contributed by MALAT1 via targeting the miRNA.

Vagus nerve stimulation optimized cardiomyocyte phenotype, sarcomere organization and energy metabolism in infarcted heart through FoxO3A-VEGF signaling

“…Similar to previous studies that reported that VEGF-A/B could readjust endothelial cells and adipocytes metabolic pathways to favor uptake, transport and oxidation of fatty acids and glucose across and into cells 22,57,58 , besides inducing stem cell differentiation into cardiomyocytes [59][60][61][62] . Our results show that VNS and ACh-induced VEGF-A primarily involved in glucose uptake while induced VEGF-B mainly participated in lipid uptake in myocytes, suggesting that inactivated FoxO3A by VNS triggered the alteration of cardiomyocytes phenotype switch and corresponding metabolic improvement targeting VEGF-A/B.…”

Extracellular Vesicle miRNAs in the Promotion of Cardiac Neovascularisation

“…The authors showed that RIC and RIC exosomes significantly improved cardiac function and blood vessel formation, and decreased collagen deposition 28 days post-MI. Wu et al ( 2020 ), demonstrated that the intramyocardial injection of EVs from ESC-derived cardiovascular progenitor cells (CVPCs) cultured under normoxia or hypoxia could significantly improve cardiac function, vascularisation and cardiomyocyte survival, and reduce fibrosis at 28 days in a mouse MI model. More importantly, EVs secreted from ESC-CVPC cultured under hypoxia had a better benefit in improving cardiac function post-MI.…”

“…Increasing evidence suggests that the interactions of lncRNAs with miRNAs play a critical role in the regulation of angiogenesis (Zhao et al, 2020 ). Moreover, lncRNAs may be selectively packaged into EVs and transferred between cells, improving cardiac function and vascularisation and reducing fibrosis post-MI (Wu et al, 2020 ). Circular RNAs (circRNAs) have also emerged as novel regulators of EC function and angiogenesis (e.g., CANRIL, CZNF292, Circ_0010729,Circ_0003575, Circ_0054633, Circ_0000109) (Zhang and Huang, 2020 ).…”

Advances in Exosomes Derived from Different Cell Sources and Cardiovascular Diseases

“…Additionally, miR-133a-cardiac progenitor cells clearly improved cardiac function in a rat myocardial infarction model by reducing fibrosis and hypertrophy and increasing vascularization and cardiomyocyte proliferation [ 44 ]. The cardiac precursor cell-secreted exosomes promote the infarct healing through the improvement of cardiomyocyte survival and angiogenesis, and the cardioprotective effects can be enhanced by hypoxia conditioning of cardiac precursor cells and are partially contributed by MALAT1 via targeting the miRNA [ 45 ]. In contrast, exosomes from cardiac progenitor cells have also been shown to stimulate cell migration [ 46 ].…”