CARMN Is an Evolutionarily Conserved Smooth Muscle Cell-Specific LncRNA That Maintains Contractile Phenotype by Binding Myocardin.
CARMN Is an Evolutionarily Conserved Smooth Muscle Cell-Specific LncRNA That Maintains Contractile Phenotype by Binding Myocardin.
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DOI：
10.1161/circulationaha.121.055949
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发表时间：
2021-12-07
期刊：
影响因子：
37.8
通讯作者：
Zhou J
中科院分区：
文献类型：
--
作者：
Dong K;Shen J;He X;Hu G;Wang L;Osman I;Bunting KM;Dixon-Melvin R;Zheng Z;Xin H;Xiang M;Vazdarjanova A;Fulton DJR;Zhou J
血管内环境稳定是由血管平滑肌细胞（VSMCs）的分化表型维持的。蛋白质编码基因的景观组成的转录组分化的血管平滑肌细胞已被深入研究，但仍有许多差距，包括非编码基因的新兴作用。我们重新分析了来自多种组织和细胞类型的大规模、公开可用的批量和scRNA-seq数据集，以鉴定富含VSMC的lncRNA。使用新型Carmn GFP敲入报告基因小鼠模型研究了新型SMC表达的lncRNA Carmn（CARdiac中胚层增强子相关非编码RNA）的体内表达模式。采用生物信息学和qRT-PCR分析来评估人和鼠血管疾病模型中VSMC表型调节期间CARMN表达的变化。在体外，通过用反义寡核苷酸敲低CARMN并在人冠状动脉SMC中通过腺病毒过表达Carmn来进行功能测定。在SMC特异性Carmn敲除小鼠中进行颈动脉损伤以评估新生内膜形成，并在大鼠颈动脉球囊损伤模型中测试逆转CARMN损失的治疗潜力。使用RNA下拉、RNA免疫沉淀和荧光素酶报告基因测定研究CARMN功能的分子机制。我们鉴定了CARMN，它最初被注释为MIR 143/145簇的宿主基因，最近报道在心脏分化中发挥作用，作为一种高度丰富和保守的SMC特异性lncRNA。Carmn GFP敲入小鼠模型的分析证实，Carmn在胚胎心肌细胞中瞬时表达，此后仅限于SMC。我们还发现Carmn的转录独立于Mir 143/145。CARMN表达在人类和小鼠模型中因血管疾病而显著降低，并在体外调节VSMC的收缩表型。在体内，SMC特异性Carmn缺失显著加剧，而Carmn过表达显著减弱，分别在小鼠和大鼠中损伤诱导的新生内膜形成。从机制上讲，我们发现Carmn物理结合的关键转录辅因子myocardin，促进其活动，从而保持收缩表型的VSMC。CARMN是一种进化上保守的SMC特异性lncRNA，在维持VSMC的收缩表型方面具有以前未被认识到的作用，并且是发现的第一个与心肌蛋白相互作用的非编码RNA。
Vascular homeostasis is maintained by the differentiated phenotype of vascular smooth muscle cells (VSMCs). The landscape of protein coding genes comprising the transcriptome of differentiated VSMCs has been intensively investigated but many gaps remain including the emerging roles of non-coding genes. We re-analyzed large-scale, publicly available bulk and scRNA-seq datasets from multiple tissues and cell types to identify VSMC-enriched lncRNAs. The in vivo expression pattern of a novel SMC expressed lncRNA, Carmn (CARdiac Mesoderm Enhancer-associated Non-coding RNA) was investigated using a novel Carmn GFP knock-in reporter mouse model. Bioinformatics and qRT-PCR analysis were employed to assess CARMN expression changes during VSMC phenotypic modulation in human and murine vascular disease models. In vitro, functional assays were performed by knocking down CARMN with antisense oligonucleotides and over-expressing Carmn by adenovirus in human coronary artery SMCs. Carotid artery injury was performed in SMC-specific Carmn knockout mice to assess neointima formation and the therapeutic potential of reversing CARMN loss was tested in a rat carotid artery balloon injury model. The molecular mechanisms underlying CARMN function were investigated using RNA pull-down, RNA immunoprecipitation and luciferase reporter assays. We identified CARMN, which was initially annotated as the host gene of the MIR143/145 cluster and recently reported to play a role in cardiac differentiation, as a highly abundant and conserved, SMC-specific lncRNA. Analysis of the Carmn GFP knock-in mouse model confirmed that Carmn is transiently expressed in embryonic cardiomyocytes and thereafter becomes restricted to SMCs. We also found that Carmn is transcribed independently of Mir143/145. CARMN expression is dramatically decreased by vascular disease in humans and murine models and regulates the contractile phenotype of VSMCs in vitro. In vivo, SMC-specific deletion of Carmn significantly exacerbated, while overexpression of Carmn markedly attenuated, injury-induced neointima formation in mouse and rat, respectively. Mechanistically, we found that Carmn physically binds to the key transcriptional cofactor myocardin, facilitating its activity and thereby maintaining the contractile phenotype of VSMCs. CARMN is an evolutionarily conserved SMC-specific lncRNA with a previously unappreciated role in maintaining the contractile phenotype of VSMCs and is the first non-coding RNA discovered to interact with myocardin.
DOI：
10.1083/jcb.109.6.2929
发表时间：
1989-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Sawtell NM;Lessard JL
通讯作者：
Lessard JL