Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia; Centre for Cardiac and Vascular Biology, The University of Queensland, Brisbane, QLD 4072, Australia.
Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia.
Cell Stem Cell. 2018 Oct 4;23(4):586-598.e8. doi: 10.1016/j.stem.2018.09.009.
Cardiac differentiation of human pluripotent stem cells (hPSCs) requires orchestration of dynamic gene regulatory networks during stepwise fate transitions but often generates immature cell types that do not fully recapitulate properties of their adult counterparts, suggesting incomplete activation of key transcriptional networks. We performed extensive single-cell transcriptomic analyses to map fate choices and gene expression programs during cardiac differentiation of hPSCs and identified strategies to improve in vitro cardiomyocyte differentiation. Utilizing genetic gain- and loss-of-function approaches, we found that hypertrophic signaling is not effectively activated during monolayer-based cardiac differentiation, thereby preventing expression of HOPX and its activation of downstream genes that govern late stages of cardiomyocyte maturation. This study therefore provides a key transcriptional roadmap of in vitro cardiac differentiation at single-cell resolution, revealing fundamental mechanisms underlying heart development and differentiation of hPSC-derived cardiomyocytes.
人多能干细胞（hPSCs）的心脏分化需要在逐步的命运转变过程中协调动态的基因调控网络，但通常产生不成熟的细胞类型，不能完全再现其成体对应物的特性，这表明关键转录网络的不完全激活。我们进行了广泛的单细胞转录组分析，以绘制 hPSC 心脏分化过程中的命运选择和基因表达程序，并确定了改善体外心肌细胞分化的策略。利用遗传功能获得和缺失方法，我们发现肥厚信号在基于单层的心脏分化过程中不能有效激活，从而阻止 HOPX 的表达及其对下游基因的激活，这些基因控制心肌细胞成熟的后期阶段。因此，这项研究提供了单细胞分辨率下体外心脏分化的关键转录路线图，揭示了心脏发育和 hPSC 衍生心肌细胞分化的基本机制。
