Strategic Center of Cell & Bio Therapy, Seoul National University Hospital, Seoul 03080, Republic of Korea; Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, Republic of Korea.
Department of Internal Medicine, Seoul National University Hospital, Seoul, Republic of Korea.
Stem Cell Reports. 2021 Apr 13;16(4):868-882. doi: 10.1016/j.stemcr.2021.03.003. Epub 2021 Apr 1.
Identifying lineage-specific markers is pivotal for understanding developmental processes and developing cell therapies. Here, we investigated the functioning of a cardiomyogenic cell-surface marker, latrophilin-2 (LPHN2), an adhesion G-protein-coupled receptor, in cardiac differentiation. LPHN2 was selectively expressed in cardiac progenitor cells (CPCs) and cardiomyocytes (CMCs) during mouse and human pluripotent stem cell (PSC) differentiation; cell sorting with an anti-LPHN2 antibody promoted the isolation of populations highly enriched in CPCs and CMCs. Lphn2 knockdown or knockout PSCs did not express cardiac genes. We used the Phospho Explorer Antibody Array, which encompasses nearly all known signaling pathways, to assess molecular mechanisms underlying LPHN2-induced cardiac differentiation. LPHN2-dependent phosphorylation was the strongest for cyclin-dependent kinase 5 (CDK5) at Tyr15. We identified CDK5, Src, and P38MAPK as key downstream molecules of LPHN2 signaling. These findings provide a valuable strategy for isolating CPCs and CMCs from PSCs and insights into the still-unknown cardiac differentiation mechanisms.
鉴定谱系特异性标志物对于理解发育过程和开发细胞疗法至关重要。在这里，我们研究了一种心肌细胞表面标志物——拉托菲林-2（LPHN2）的功能，它是一种黏附 G 蛋白偶联受体，在心脏分化中起作用。在小鼠和人多能干细胞（PSC）分化过程中，LPHN2 选择性地在心脏祖细胞（CPCs）和心肌细胞（CMCs）中表达；用抗 LPHN2 抗体进行细胞分选可促进 CPCs 和 CMCs 高度富集的群体的分离。Lphn2 敲低或敲除 PSCs 不表达心脏基因。我们使用了涵盖几乎所有已知信号通路的 Phospho Explorer Antibody Array 来评估 LPHN2 诱导心脏分化的分子机制。LPHN2 依赖性磷酸化在 Tyr15 处对周期蛋白依赖性激酶 5（CDK5）最强。我们鉴定出 CDK5、Src 和 P38MAPK 是 LPHN2 信号的关键下游分子。这些发现为从 PSCs 中分离 CPCs 和 CMCs 提供了有价值的策略，并深入了解了仍未知的心脏分化机制。
