非编码RNA在心肌细胞形成中的作用与调控机制

To obtain cardiomyocytes by direct differentiation and transdifferentiation is the most promising frontier for the treatment of heart diseases, but the low efficiency and safety of transdifferentiation are still controversial. Non-coding RNA plays critical function in cell fate transition, but it is not clear for the exact function of ncRNA and ncRNA-mediated regulatory network in cardiomyocyte generation. We will use the in vitro and in vivo model of cardiomyocyte generation, and our expertise in the function and mechanism research of ncRNA in regulating cell fate. In combination with the bioinformatics analysis and systematic screening for cardiomyocyte-specific ncRNA that may potentially target myocardial cell related transcription factors and signaling pathways, we will further clarify the function of specific ncRNA such as lncRNA-TPB in cardiomyocyte generation. The specific regulatory mechanism of ncRNA in cardiomyocyte generation will be further elucidated by molecular experiments including luciferase reporter gene, RNA pull-down, RIP and so on. On this basis, the newly discovered ncRNA will be introduced into the in vivo transdifferentiation model, and the effects of key ncRNA on the efficiency of in vivo transdifferentiation, and cardiac function recovery will also be investigated. The completion of this project will further enrich the ncRNA-mediated epigenetic-specific regulatory network in cardiomyocyte generation, provide important basis for obtaining high quality cardiomyocytes with high efficiency, and potential target for early and safe application of cardiomyocytes in disease treatment.

定向分化和转分化获得心肌细胞是治疗心脏疾病的前沿研究，但分化的低效和安全性仍受争议。非编码RNA（ncRNA）在细胞命运转变中具有重要作用，但心肌细胞形成中ncRNA的作用及其介导的调控网络尚不明确。我们将利用心肌细胞形成的体内外模型与ncRNA调控细胞命运研究方面的特长，结合生物信息分析系统筛查可能调控心肌细胞关键转录因子和信号通路的心肌特异性ncRNA，并进一步明确lncRNA-TPB等在心肌细胞形成中的功能，通过报告基因、RNA pull-down、RIP等实验阐明ncRNA在心肌细胞形成中的具体调控机制。在此基础上，将新发现的ncRNA介入到心肌细胞体内转分化模型中，分析关键ncRNA对体内心肌细胞转分化效率和心脏功能回复等方面的影响。本项目的完成，将更加丰富心肌细胞形成中ncRNA介导的表观遗传特异调控网络，为高效获得高质量心肌细胞及其早日安全用于疾病治疗提供重要依据和新靶点。

心血管疾病是全球致死的最主要原因之一，具有高发病率和高死亡率的特点。如何高效地获得高质量的心肌细胞对于治疗心血管疾病具有重要意义。干细胞具有自我更新和多向分化潜能，在再生医学中具有重要作用。借助干细胞体外定向心肌细胞分化体系可获得足量的心肌细胞，但是干细胞定向分化形成心肌细胞的低效和安全性仍受争议。非编码RNA在细胞命运转变中具有重要作用，但心肌细胞形成中的非编码RNA作用及其调控网络尚不明确。本项目立项以来，筛查并鉴定得到了心肌细胞形成密切相关的非编码RNA类群，确定了特定非编码RNA分子在心肌细胞形成中的功能，阐明了特定非编码RNA分子与转录因子和关键信号通路间的具体作用关系，揭示了多个非编码RNA在心肌细胞形成中的调控机制，解析了心肌细胞形成中由非编码RNA介导的表观遗传特异性调控网络，为高质量心肌细胞的获得以及心脏类疾病的诊疗提供了重要理论依据和潜在的非编码RNA靶点。项目实施取得了突破性成果，圆满完成了研究任务和目标，在国外重要杂志发表标注本项目基金号的学术论文4篇，培养毕业研究生6名。具体成果如下：1）揭示了LncCMRR在心肌细胞形成中的新功能与表观遗传调控机制；2）揭示了miR-184介导Wnt3信号通路调控干细胞向心肌中胚层分化的新功能；3）阐明了linc1557在干细胞多能性退出和分化起始中的新功能与新机制；4）阐明了lncCmarr/miR-540-3p促进心肌细胞成熟的新功能与新机制。