Downregulation of LGR5 Expression Inhibits Cardiomyocyte Differentiation and Potentiates Endothelial Differentiation from Human Pluripotent Stem Cells

Abstract

Summary: Understanding molecules involved in differentiation of human pluripotent stem cells (hPSCs) into cardiomyocytes and endothelial cells is important in advancing hPSCs for cell therapy and drug testing. Here, we report that LGR5, a leucine-rich repeat-containing G-protein-coupled receptor, plays a critical role in hPSC differentiation into cardiomyocytes and endothelial cells. LGR5 expression was transiently upregulated during the early stage of cardiomyocyte differentiation, and knockdown of LGR5 resulted in reduced expression of cardiomyocyte-associated markers and poor cardiac differentiation. In contrast, knockdown of LGR5 promoted differentiation of endothelial-like cells with increased expression of endothelial cell markers and appropriate functional characteristics, including the ability to form tube-like structures and to take up acetylated low-density lipoproteins. Furthermore, knockdown of LGR5 significantly reduced the proliferation of differentiated cells and increased the nuclear translocation of β-catenin and expression of Wnt signaling-related genes. Therefore, regulation of LGR5 may facilitate efficient generation of cardiomyocytes or endothelial cells from hPSCs. : In this article, Xu and colleagues demonstrate that LGR5 (leucine-rich repeat-containing G-protein-coupled receptor 5) plays a critical role in cardiomyocytes and endothelial differentiation from human pluripotent stem cells. Expression of LGR5 is transiently upregulated during the early stage of cardiomyocyte differentiation, and knockdown of LGR5 results in reduced cardiomyocyte differentiation but increased endothelial differentiation. Keywords: cardiomyocytes, differentiation, embryonic stem cells, endothelial cells, gene expression, induced pluripotent stem cells, LGR5, mesoderm patterning, proliferation, Wnt signaling