长链非编码RNAs调节BMSCs向心肌分化的作用与机制

Low differentiation rate and pro-arrhythmic potential of transplanted BMSCs in heart diseases hamper broad application of BMSCs transplantation in clinics. Promoting cardiomyocyte differentiation of transplanted BMSCs is a key approach to overcoming this problem. Latest studies showed that lncRNAs are involved in a variety of biological process such as proliferation and apoptosis. However, the role and mechanisms of LncRNAs in cardiac differentiation of BMSCs remain unclear. Our preliminary results demonstrated that Zfas1 and GAS5 were altered in this process, and Zfas1 and GAS5 played a regulatory role in cardiac differentiation of BMSCs. In this project we employ patch clamp, Luciferase, EMSA, etc to study the following contents: (1) Whether Zfas1 and GAS5 can synergistically promote BMSCs differentiation to cardiomyocytes; (2) Identify Zfas1/GAS5-miRNAs/Lin28-Target genes as signal pathway of Zfas1 and GAS5 in regulating BMSCs differentiation; (3) Clarify that Zfas1 and GAS5 overexpression can enhance the efficacy and safety of BMSCs transplantation, which will provide new strategy for BMSCs-based heart repair.

骨髓基质干细胞（BMSCs）心脏移植后向心肌分化率低和潜在致心律失常风险制约了临床上BMSCs移植的广泛开展，如何促进BMSCs向心肌分化是解决这一问题的关键。新近研究发现长链非编码RNAs（LncRNAs）调控细胞增殖、凋亡等病理生理过程，但LncRNAs在BMSCs向心肌分化过程中的作用和机制仍不清楚。本项目前期实验发现LncRNAs-Zfas1和GAS5在BMSCs向心肌分化过程中变化显著，并首次揭示它们对BMSCs向心肌分化具有调控作用。本项目拟运用膜片钳、报告基因、EMSA等实验技术研究（1）Zfas1和GAS5是否具有协同促BMSCs向心肌分化作用；（2）揭示Zfas1/GAS5-miRNAs/Lin28-靶基因通路在BMSCs分化中的调控作用；（3）验证Zfas1和GAS5过表达可增强BMSCs移植的疗效与安全性，为临床BMSCs心脏移植提供新思路和优化策略。

干细胞心脏移植是近年来的研究热点，但是干细胞移植后向心肌分化率比率低及潜在致心律失常风险制约了临床上干细胞移植的广泛开展，如何促进干细胞向心肌分化是解决这一问题的关键。大量研究揭示长链非编码RNAs（LncRNAs）参与了细胞增殖、凋亡等病理生理过程，但LncRNAs在干细胞向心肌分化过程中的作用和机制仍不清楚。本项目的研究成果包括：（1）通过研究我们发现LncRNA ZFAS1和GAS5在干细胞向心肌分化过程中表达发生变化；LncRNA ZFAS1在干细胞向心肌分化过程中表达增加，沉默LncRNA ZFAS1可明显抑制干细胞向心肌分化，LncRNA ZFAS1可调控中胚层关键分子BRACHYURY和心肌前体细胞关键分子TBX-5表达，而对OCT-4无明显作用。进一步我们发现干细胞敲减LncRNA ZFAS1后ZNFX1表达明显增加，沉默ZNFX1可以减弱ZFAS1 siRNA对干细胞向心肌分化的抑制作用；（2）还发现LncRNA GAS5在干细胞向心肌分化过程中下调，进一步研究揭示GAS5过表达可以抑制干细胞向心肌分化。通过上述研究结果我们发现揭LncRNA ZFAS1和GAS5在干细胞向心肌分化过程中具有重要作用，这一研究发现不仅揭示了LncRNAs可调控干细胞向心肌细胞分化，而且有望为提高临床干细胞移植疗效提供新策略。