Time-regulated transcripts with the potential to modulate human pluripotent stem cell-derived cardiomyocyte differentiation [miRNA]

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CM) are a promising disease model, although hPSC-CMs are not fully mature cardiomyocytes. We examined exponential differentially expressed miRNA/gene to identify critical time-regulated transcripts associated with hPSC-CM differentiation. We uncovered 324 interactions among 29 differentially expressed genes and 51 miRNAs from 20.543 transcripts during the 120 days of hPSC-CM differentiation. We found 16 genes and 26 miRNAs with an inverse pattern of expression (Pearson R-values < -0.5) validated using several human and mouse databases. We further validated these findings using two hPSC-CM lines and seven sampling times over a 30-day protocol and observed 16 inverse interactions among eight genes and 12 miRNAs (Person R-values < -0.5) changing over time. Finally, we tested the top eight miRNAs by adding miRNA mimics to differentiating hPSC-CMs and found that they influenced proliferation and maturation phenotypes. These time-regulated transcripts appear to regulate single or multiple pathways affecting cardiac differentiation. microRNA expression array of human pluripotent stem cells (hPSCs), hPSCs progenitor cardiomyocytes, and hiPSC early cardiomyocytes at different time-points of differentiation were analyzed in duplicates at 0, 2, 4, 6, 8, 15 and 30 days of maturation microRNA extracted from hPSC commited to cardiomyocytes differentiation protocol in different time points of development