LncRNA CRRL作为ceRNA调控miRNAs在心梗后心肌再生中的作用及机制研究

Gene targeted therapy is a new promising strategy for endogenous myocardial regeneration after myocardial infarction. Recent studies found that long non-coding RNA (lncRNA) played important roles in heart development, but the role in endogenous myocardial regeneration remains unclear. We previously analyzed the publicly available RNA-seq data of human fetal (mid-term) and normal adult cardiac tissues and found that a large number of lncRNAs were differentially expressed. Among them, we found a key lncRNA (we named it CRRL) which was likely involved in cardiac regeneration, and validated it could promote myocardial regeneration through pre-experiment. Further bioinformatics predict CRRL may bind with multiple miRNAs which have proved to involve in myocardial regeneration. Therefore, we assume: CRRL could functions as a competing endogenous RNA by sponging miRNAs in post-infarction endogenous myocardial regeneration. In this study, we aim to use the gain-of-function and loss-of-function method to explore whether CRRL could promote cardiac proliferation in both vivo and vitro, and attenuate post-infarction cardiac remodeling. Finally, the mechanism involved in was determined by luciferase assay and RNA pull down assay. Our study may provide a new gene target for therapy heart failure following myocardial infarction.

基因靶向治疗促进内源性心肌再生是防治心梗后心衰十分有前景的新策略。新近研究发现长链非编码RNA(lncRNA)在心脏发育中起着非常重要作用，然而其能否促进内源性心肌再生尚不清楚。我们前期研究通过比较人胎儿中期和成年心脏的lncRNA表达谱的差异，筛选并预实验验证具有促进心肌再生潜能的lncRNA (命名为CRRL)，通过生物信息方法预测CRRL能与多个参与心肌再生的miRNAs结合。因此，我们假设：CRRL作为ceRNA(competing endogenous RNAs)调控miRNAs介导的内源性心肌再生信号转导通路。本项目拟利用功能获得/缺失的方法在细胞及动物水平验证CRRL对心肌再生及心梗后心室重构的影响；通过荧光素酶实验及RNA pull down 实验探索CRRL能否作为ceRNA调控miRNAs介导的心肌再生信号转导通路。本项目将为防治心梗后心衰提供潜在基因新靶点和理论依据。

基因靶向治疗促进内源性心肌再生是防治心梗后心衰十分有前景的新策略。长链非编码RNA(lncRNA)在心脏发育中起着非常重要作用，然而其能否促进内源性心肌再生尚不清楚。我们通过比较人胎儿中期和成年心脏的lncRNA表达谱的差异，筛选出成年人心脏表达上调的lncRNA CRRL。生物信息学分析预测CRRL参与了细胞增殖的负调控。CRRL具有物种保守性且其表达量随着年龄逐渐减低。体内外实验发现，敲除CRRL能有效促进7天大鼠心肌细胞增殖。体内敲除CRRL能显著减少成年鼠和7天鼠心梗后梗死面积，有效改善成年大鼠心梗后心功能。此外，细胞亚定位结果发现CRRL主要表达在细胞浆内，进一步的机制研究发现CRRL能直接与miR-199a-3p结合, 并作为竞争性内源RNA(ceRNA)下调miR-199a-3p的靶基因Hopx来抑制心肌再生。因此，本项目研究结果证明人源性lncRNA(CRRL)通过CRRL/miR-199a-3p/Hopx轴调控心肌再生从而影响心梗后心功能。本项目的研究成果提示CRRL可以作为促进心肌再生的新型有效基因靶标，为临床防治心梗后心力衰竭提供一种潜在基因靶向治疗策略。