Introduction: Studies have shown that mRNA splicing is regulated by scaRNAs and plays a significant role in mammalian cardiac development and differentiation, but the potential contribution to human in vitro differentiation of iPSCs into CMCs remains unknown. In this study, the comparison of scaRNA20 modified and unmodified iPSCs derived-CMCs (iCMCs) will allow us to understand the role of scaRNAs in cardiac differentiation.
Hypothesis: We hypothesize that CMCs modulated with scaRNA20 plays an important role in iPSCs-derived CMC differentiation.
Methods and Results: Recently, our RNA sequencing analysis of iCMCs indicated that scaRNA20 plays an important role in CMC differentiation. We observed that si-scaRNA20 knockdown iPSCs underwent CMCs differentiation. They failed to grow as mature functional CMCs (Fig. A) and the number of beating clusters were reduced compared to the scrambled control cells (Fig. B). Our FACS data showed that there was a significant reduction in the expression of Gata4 and CTT in the CMC-derived from iPSCs kockdown with si-scaRNA20-1 but not in those with si-scaRNA-2 (Fig. C). Similarly, our FACS data from scaRNA20 overexpressing iPSCs differentiated into CMCs showed a significant increase in the number of Gata4- and CTT-expressing cells, suggesting the role of scaRNA20 in CMC differentiation.
Conclusions: We report for the first time the role scaRNA20 in the regulation of in-vitro cardiac development.