Role of alpha-and beta-adrenergic receptors in cardiomyocyte differentiation from murine-induced pluripotent stem cells
Role of alpha-and beta-adrenergic receptors in cardiomyocyte differentiation from murine-induced pluripotent stem cells
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α-和β-肾上腺素能受体在小鼠诱导多能干细胞分化为心肌细胞中的作用
DOI：
10.1111/cpr.12310
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发表时间：
2017
影响因子：
8.5
通讯作者：
Zheng Qiang-Sun
中科院分区：
文献类型：
--
作者：
Li Xiao-Li;Zeng Di;Chen Yan;Ding Lu;Li Wen-Ju;Wei Ting;Ou Dong-Bo;Yan Song;Wang Bin;Zheng Qiang-Sun
目的诱导多能干细胞（iPSC）衍生的心肌细胞是再生性心脏病治疗的一种有前途的细胞来源，但其低分化效率和高细胞异质性限制了其应用的进展。先前的研究已经证明了肾上腺素能受体（AR）在干细胞分化后的表达;然而，AR在干细胞的命运特化中的作用，特别是在心肌细胞分化和发育中，尚未被表征。材料和方法小鼠诱导的多能干细胞（miPSC）在悬滴中培养以形成胚状体，然后将其细胞分化为心肌细胞。为了确定AR是否调节miPSC分化为心脏谱系，AR激动剂肾上腺素（EPI）对miPSC分化和潜在信号传导机制的影响，进行了评估。结果EPI治疗，强烈增强miPSC心脏分化，如心脏特异性标志物GATA 4，Nkx 2. 5和Tnnt 2的表达水平增加所示。虽然β-AR信号传导是心肌细胞中最重要的信号传导途径，但EPI增强的心脏分化更多地依赖于α-AR信号传导而不是β-AR信号传导。此外，用特异性激动剂选择性激活α1-AR信号传导诱导了剧烈的心肌细胞分化，而选择性激活α2-或β-AR信号传导分别诱导了无分化或较少的分化。EPI和α1-AR依赖的miPSC心肌细胞分化是通过特异性促进CPC增殖、MEK-ERK 1/2通路和调节miPS细胞周期进程而发生的。结论这些结果表明，AR的激活，特别是α1-AR的激活，通过MEK-ERK 1/2信号传导促进miPSC分化为心肌细胞系。
ObjectivesInduced pluripotent stem cell (iPSC)‐derived cardiomyocytes are a promising source of cells for regenerative heart disease therapies, but progress towards their use has been limited by their low differentiation efficiency and high cellular heterogeneity. Previous studies have demonstrated expression of adrenergic receptors (ARs) in stem cells after differentiation; however, roles of ARs in fate specification of stem cells, particularly in cardiomyocyte differentiation and development, have not been characterized.Materials and methodsMurine‐induced pluripotent stem cells (miPSCs) were cultured in hanging drops to form embryoid bodies, cells of which were then differentiated into cardiomyocytes. To determine whether ARs regulated miPSC differentiation into cardiac lineages, effects of the AR agonist, epinephrine (EPI), on miPSC differentiation and underlying signalling mechanisms, were evaluated.ResultsTreatment with EPI, robustly enhanced miPSC cardiac differentiation, as indicated by increased expression levels of cardiac‐specific markers,GATA4,Nkx2.5andTnnt2. Although β‐AR signalling is the foremost signalling pathway in cardiomyocytes, EPI‐enhanced cardiac differentiation depended more on α‐AR signalling than β‐AR signalling. In addition, selective activation of α1‐AR signalling with specific agonists induced vigorous cardiomyocyte differentiation, whereas selective activation of α2‐ or β‐AR signalling induced no or less differentiation, respectively. EPI‐ and α1‐AR‐dependent cardiomyocyte differentiation from miPSCs occurred through specific promotion of CPC proliferationviathe MEK‐ERK1/2 pathway and regulation of miPS cell‐cycle progression.ConclusionsThese results demonstrate that activation of ARs, particularly of α1‐ARs, promoted miPSC differentiation into cardiac lineagesviaMEK‐ERK1/2 signalling.