Abstract 11919: Role of microRNA-1a (miR-1a) in the Differentiation of Induced Pluripotent Stem Cells (iPSCs) to Endothelial Cells (ECs)
Background: miR-1a has been reported to promote cardiac differentiation of iPSCs. DNA methylatransferase 3a (DNMT3a) plays an important role in neovascularization by mediating the methylation of endothelial specific genes. However, the miR-1/ DNMT3a interaction in the endothelial differentiation is not well understood.
Methods: Mouse iPSCs were cultured for 14 days in differentiation media to yield ECs. miR-1a, DNMT3a, EC marker (VE-cadherin), and stem cell marker (Oct4) expression were analyzed by qRT-PCR. The effect of miR-1a on DNMT3a was analyzed by a luciferase 3’UTR reporter. EC differentiation was assessed by acetylated-LDL uptake and vWF staining. Additionally, the effect of 5-azacytidine, a DNMT inhibitor, on VE-cadherin, eNOS, and GATA2 was also analyzed by qRT-PCR.
Results: Compared to the native iPSCs, miR-1a and VE-cadherin expression were increased in iPSCs cultured for 14 days, while DNMT3a and Oct4 expression were decreased (Fig.1). The administration of a miR-1a analog reduced both DNMT3a expression and the luciferase 3'UTR activity. miR-1a enhanced EC differentiation of iPSCs as determined by ac-LDL uptake and vWF staining. Inhibition of DNMT activity by 5-azacytidine augmented the expression of VE-cadherin, eNOS, and GATA2 in iPSCs.
Conclusion: miRNA-1a enhances EC differentiation of iPSCs by targeting DNMT3a and it is a novel strategy for generating cardiac progenitors for cell based therapy.
- © 2012 by American Heart Association, Inc.