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Exosomes secreted by the CCND2OECMs promote proliferation and hypoxia tolerance in cultured CMs

ABSTRACT: CCND2OEExos (exosomes from CCND2-overexpressing hiPSC-derived cardiomyocytes (CCND2OECMs)) were found to be able to promote proliferation of not only cardiomyocytes, but also human umbilical vein endothelial cells (HUVEC) and human vascular smooth muscle cells (HVSMC) in vitro, as indicated by the large number of ki67+ and PH3+ HUVECs and SMCs in the CCND2OEExos-treated group, relative to those in the vehicle only- or CCND2WTExos-treated groups. A comparative bulk sequence analysis of miRNAs from CCND2WTExos and CCND2OEExos (exosomes from wild type hiPSC-derived cardiomyocytes) was performed to identify some underlying mechanisms about proliferation induction.

ORGANISM(S): Homo sapiens

PROVIDER: GSE161046 | GEO | 2022/04/26

REPOSITORIES: GEO

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Project description:ABSTRACT Background: Viral myocarditis is a life-threatening illness that may lead to heart failure or cardiac arrhythmias. This study examined whether human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) could be used to model the pathogenic processes of coxsackievirus-induced viral myocarditis and to screen antiviral therapeutics for efficacy. Methods and Results: Human iPSC-CMs were infected with a luciferase-expressing mutant of the coxsackievirus B3 strain (CVB3-Luc). Brightfield microscopy, immunofluorescence, and calcium imaging were used to characterize virally infected hiPSC-CMs. Viral proliferation on hiPSC-CMs was subsequently quantified using bioluminescence imaging. For drug screening, select antiviral compounds including interferon beta 1 (IFNβ1), ribavirin, pyrrolidine dithiocarbamate (PDTC), and fluoxetine were tested for their capacity to abrogate CVB3-Luc proliferation in hiPSC-CMs in vitro. The ability of some of these compounds to reduce CVB3-Luc proliferation in hiPSC-CMs was consistent with the reported drug effects in previous studies. Finally, mechanistic analyses via gene expression profiling of hiPSC-CMs infected with CVB3-Luc revealed an activation of viral RNA and protein clearance pathways within these hiPSC-CMs after IFNβ1 treatment. Conclusions: This study demonstrates that hiPSC-CMs express the coxsackievirus and adenovirus receptor, are susceptible to coxsackievirus infection, and can be used to confirm antiviral drug efficacy. Our results suggest that the hiPSC-CM/CVB3-Luc assay is a sensitive platform that could be used to screen novel antiviral therapeutics for their effectiveness in a high-throughput fashion. For this experiment, human induced pluripotent stem cell derived cardiomyocytes were infected with coxsackievirus at multiplicity of infection (MOI) of 5 for 8 hours. Cells were treated with and without interferon beta 1 in order to determine if treatment activates antiviral response genes and/or viral clearance pathways. 4 total samples (2 for each condition) were analyzed

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Exosomes secreted by the CCND2OECMs promote proliferation and hypoxia tolerance in cultured CMs

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