Comparison of Non-Coding RNAs in Exosomes and Functional Efficacy of Human Embryonic Stem Cell- versus Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Abstract

Background Both human embryonic stem cell-derived cardiomyocytes (ESC-CMs) and human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) can serve as unlimited cell sources for cardiac regenerative therapy. However, the functional equivalency between human ESC-CMs and iPSC-CMs for cardiac regenerative therapy has not been demonstrated. Here we performed a head-to-head comparison of ESC-CMs and iPSC-CMs in their ability to restore cardiac function in a rat myocardial infarction (MI) model as well as their exosomal secretome. Methods and Results Human ESCs and iPSCs were differentiated into cardiomyocytes using small molecule inhibitors. Fluorescence-activated cell sorting (FACS) analysis confirmed ~85% and ~83% of CMs differentiated from ESCs and iPSCs, respectively, were positive for cardiac troponin T. At a single-cell level, both cell types displayed similar calcium handling and electrophysiological properties, with gene expression comparable to the human fetal heart marked by striated sarcomeres. Sub-acute transplantation of ESC-CMs and iPSC-CMs into nude rats post-MI improved cardiac function, which was associated with increased expression of angiogenic genes in vitro following hypoxia. Profiling of exosomal microRNAs (miRs) and long non-coding RNAs (lncRNAs) revealed that both groups contain an identical repertoire of miRs and lncRNAs, including some that are known to be cardioprotective. Conclusions We demonstrate for the first time that both ESC-CMs and iPSC-CMs can facilitate comparable cardiac repair. This is advantageous because unlike allogeneic ESC-CMs used in therapy, autologous iPSC-CMs could potentially avoid immune rejection when used for cardiac cell transplantation in the future.

Utilization of Human Induced Pluripotent Stem Cells for Cardiac Repair

“…Khan et al (2015) reported that ESCsderived exosomes, after being directly injected to infarcted mouse hearts, promote cardiac pro-angiogenesis and cardiomyocyte survival, improve cardiac function and reduce fibrosis after myocardial infarction. More recent papers confirmed the therapeutic effects of exosome in other types of myocardial injuries Singla et al, 2019) and explored the potential mechanism underlying the beneficial effects (Lee et al, 2017). The therapeutic delivery of exosomes through the circulation system by using miRNAs mimics or synthetic exosomes as cargos and vehicles have been widely studied.…”

Beneficial effects of exosomes secreted by cardiac-derived progenitor cells and other cell types in myocardial ischemia

“…These exosomes transported several upregulated miRNAs, including miR-92a which has been associated with antiapoptotic, proangiogenic, and antifibrotic effects. Lee et al (79) compared ESC-derived cardiomyocytes and iPSC-derived cardiomyocytes with respect to their ability to restore cardiac function in a rat MI model, as well as their exosomal secretome. Both cell types injected into nude rats post-MI comparably improved cardiac function.…”

Immunoregulatory Effects of Stem Cell-Derived Extracellular Vesicles on Immune Cells

“…Extracellular vesicles (EVs), now identified as a novel apparatus of intercellular communication, did not garner significant attention previously, although they are currently sought after as a topic of research. EVs have a diameter ranging between 50 and 2,000 nm with a bilayer lipid membrane (1) and comprise parental cell-derived active cargos such as lipids (2), proteins (2,3), nucleic acids [(DNAs) (4), mRNA (5,6), microRNAs (miRNAs) (4), non-coding RNAs (7)] and organelles (4,8). Emerging evidence indicates that double-stranded DNA, DNA-binding histones, and certain miRNAs are not associated with small EVs such as exosomes (9).…”