长链非编码 RNA CARMA 可调控心肌细胞分化。

The conserved long non-coding RNA CARMA regulates cardiomyocyte differentiation.

机构信息

Department of Molecular Genetics, Faculty of Biological Sciences Tarbiat Modares University, Tehran, Iran.

Experimental Cardiology Unit, Division of Cardiology, Department of Cardiovascular Medicine, University of Lausanne Medical School, Lausanne, Switzerland.

出版信息

Cardiovasc Res. 2022 Jul 27;118(10):2339-2353. doi: 10.1093/cvr/cvab281.

DOI:10.1093/cvr/cvab281

PMID:34459880

Abstract

AIMS

Production of functional cardiomyocytes from pluripotent stem cells requires tight control of the differentiation process. Long non-coding RNAs (lncRNAs) exert critical regulatory functions in cell specification during development. In this study, we designed an integrated approach to identify lncRNAs implicated in cardiogenesis in differentiating human embryonic stem cells (ESCs).

METHODS AND RESULTS

We identified CARMA (CARdiomyocyte Maturation-Associated lncRNA), a conserved lncRNA controlling cardiomyocyte differentiation and maturation in human ESCs. CARMA is located adjacent to MIR-1-1HG, the host gene for two cardiogenic miRNAs: MIR1-1 and MIR-133a2, and transcribed in an antisense orientation. The expression of CARMA and the miRNAs are negatively correlated, and CARMA knockdown increases MIR1-1 and MIR-133a2 expression. In addition, CARMA possesses MIR-133a2 binding sites, suggesting the lncRNA could be also a target of miRNA action. Upon CARMA down-regulation, MIR-133a2 target protein-coding genes are coordinately down-regulated. Among those, we found RBPJ, the gene encoding the effector of the NOTCH pathway. NOTCH has been shown to control a binary cell fate decision between the mesoderm and the neuroectoderm lineages, and NOTCH inhibition leads to enhanced cardiomyocyte differentiation at the expense of neuroectodermal derivatives. Interestingly, two lncRNAs, linc1230 and linc1335, which are known repressors of neuroectodermal specification, were found up-regulated upon Notch1 silencing in ESCs. Forced expression of either linc1230 or linc1335 improved ESC-derived cardiomyocyte production. These two lncRNAs were also found up-regulated following CARMA knockdown in ESCs.

CONCLUSIONS

Altogether, these data suggest the existence of a network, implicating three newly identified lncRNAs, the two myomirs MIR1-1 and MIR-133a2 and the NOTCH signalling pathway, for the coordinated regulation of cardiogenic differentiation in ESCs.

摘要

目的

从多能干细胞中产生功能性心肌细胞需要严格控制分化过程。长非编码 RNA（lncRNA）在发育过程中细胞特化中发挥关键的调节功能。在这项研究中，我们设计了一种综合方法来鉴定参与人胚胎干细胞（hESC）分化为心肌细胞的 lncRNA。

方法和结果

我们鉴定出 CARMA（心肌细胞成熟相关 lncRNA），这是一种在 hESC 中控制心肌细胞分化和成熟的保守 lncRNA。CARMA 位于 MIR-1-1HG 附近，后者是两个心肌生成 miRNA：MIR1-1 和 MIR-133a2 的宿主基因，并以反义方向转录。CARMA 和 miRNA 的表达呈负相关，CARMA 敲低增加 MIR1-1 和 MIR-133a2 的表达。此外，CARMA 具有 MIR-133a2 的结合位点，表明该 lncRNA也可能是 miRNA 作用的靶标。CARMA 下调后，MIR-133a2 的靶蛋白编码基因协同下调。在这些基因中，我们发现了 RBPJ，它编码 NOTCH 通路的效应因子。已经表明 NOTCH 控制中胚层和神经外胚层谱系之间的二元细胞命运决定，NOTCH 抑制导致以牺牲神经外胚层衍生物为代价增强心肌细胞分化。有趣的是，在 hESC 中沉默 Notch1 时，发现两个已知的神经外胚层特化抑制子 lncRNA，linc1230 和 linc1335 上调。强制表达这两种 lncRNA 均可提高 ESC 衍生的心肌细胞产量。在 hESC 中 CARMA 敲低后，这两种 lncRNA 也发现上调。