Distinct roles of microRNA-1 and -499 in ventricular specification and functional maturation of human embryonic stem cell-derived cardiomyocytes.
Distinct roles of microRNA-1 and -499 in ventricular specification and functional maturation of human embryonic stem cell-derived cardiomyocytes.
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DOI：
10.1371/journal.pone.0027417
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Li RA
中科院分区：
文献类型：
--
作者：
Fu JD;Rushing SN;Lieu DK;Chan CW;Kong CW;Geng L;Wilson KD;Chiamvimonvat N;Boheler KR;Wu JC;Keller G;Hajjar RJ;Li RA
microRNAs（miRs）负调控转录，是正常心脏发育和心力衰竭发病机制的重要决定因素。尽管在小鼠研究中获得了重要的知识，但它们在人类（h）心脏中的功能作用仍然难以捉摸。我们假设在心脏分化中起重要作用的miR在分化、发育和终末成熟的人心肌细胞（CM）中差异表达。作为第一步，我们绘制了人（h）胚胎干细胞（ECs）、hECs衍生（hE）、胎儿（hF）和成人（hA）心室（V）CM的miR谱。63个miR在hESCs和hE-VCM之间差异表达。其中，29个，包括miR-302和-371/372/373簇，与多能性相关并在hESC中独特表达。在hE-VCM中差异表达的其余miR中，23个在hF-和hA-VCM中继续高度表达，其中miR-1、-133和-499显示出最大的倍数差异;其他如miR-let-7a、-let-7 b、-26b、-125 a和-143是非心脏特异性的。在功能上，LV-miR-499转导hESC衍生的心血管祖细胞显着增加了hE-VCM的产量（从对照的48%增加到72%; p<0.05）和收缩蛋白表达，而不影响其电生理特性（p>0.05）。相比之下，LV-miR-1转导不会使hE-VCM的产量产生偏差（p>0.05），但会降低APD和超极化RMP/MDP，这是由于Ito、IKs和IKr增加以及If降低（p<0.05），这是功能成熟的标志。此外，LV-miR-1而不是-499增强了未成熟Ca 2+瞬时幅度和动力学。进行分子途径分析以进一步了解。我们的结论是，miR-1和-499发挥不同的作用，在心脏分化的hESCs在上下文依赖的方式。虽然miR-499促进hESC的心室特化，但miR-1用于促进电生理成熟。
MicroRNAs (miRs) negatively regulate transcription and are important determinants of normal heart development and heart failure pathogenesis. Despite the significant knowledge gained in mouse studies, their functional roles in human (h) heart remain elusive. We hypothesized that miRs that figure prominently in cardiac differentiation are differentially expressed in differentiating, developing, and terminally mature human cardiomyocytes (CMs). As a first step, we mapped the miR profiles of human (h) embryonic stem cells (ESCs), hESC-derived (hE), fetal (hF) and adult (hA) ventricular (V) CMs. 63 miRs were differentially expressed between hESCs and hE-VCMs. Of these, 29, including the miR-302 and -371/372/373 clusters, were associated with pluripotency and uniquely expressed in hESCs. Of the remaining miRs differentially expressed in hE-VCMs, 23 continued to express highly in hF- and hA-VCMs, with miR-1, -133, and -499 displaying the largest fold differences; others such as miR-let-7a, -let-7b, -26b, -125a and -143 were non-cardiac specific. Functionally, LV-miR-499 transduction of hESC-derived cardiovascular progenitors significantly increased the yield of hE-VCMs (to 72% from 48% of control; p<0.05) and contractile protein expression without affecting their electrophysiological properties (p>0.05). By contrast, LV-miR-1 transduction did not bias the yield (p>0.05) but decreased APD and hyperpolarized RMP/MDP in hE-VCMs due to increased Ito, IKs and IKr, and decreased If (p<0.05) as signs of functional maturation. Also, LV-miR-1 but not -499 augmented the immature Ca2+ transient amplitude and kinetics. Molecular pathway analyses were performed for further insights. We conclude that miR-1 and -499 play differential roles in cardiac differentiation of hESCs in a context-dependent fashion. While miR-499 promotes ventricular specification of hESCs, miR-1 serves to facilitate electrophysiological maturation.
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