CircRNA-79530与TWIST形成正反馈回路调节肾纤维化EMT的分子机制研究

TWIST, the core transcription factor involved in epithelial-mesenchymal transition (EMT), attenuates renal fibrosis and prevents renal failure by inhibiting EMT progression. In our previous studies, circRNA-79530 (circ79530) can form a positive feedback with TWIST to adjust the TWIST expression, and its concentration is closely related to the EMT process. Those data indicates circ79530 can regulate the EMT in renal fibrosis by TWIST. This project will test the relationship between circ79530 and TWIST, observe the function of circ79530 in the EMT process based on EMT cellular model, research the mechanism of circ79530 on EMT in renal fibrosis by gene knock out, study the relationship between circ79530, TWIST and EMT in renal fibrosis based on renal fibrosis model and clinical samples in vivo. This study will discuss function of the positive feedback loop of circ79530 and TWIST on EMT in renal fibrosis for the first time. Because circRNA is more stable and detectable, this method is of important strategic significance and application value in the long-term monitoring and effective control for the renal fibrosis.

TWIST作为调控肾小管上皮细胞间充质转化（EMT）的核心转录因子，通过激活EMT促进肾纤维化引发肾衰竭。我们前期工作发现circRNA-79530(circ79530)通过与TWIST形成正反馈回路调节TWIST表达，其含量与EMT进程密切相关，提示circ79530是肾纤维化EMT过程中的重要调控因子。本项目将明确circ79530与TWIST的调控关系，并基于EMT细胞模型观察circ79530在EMT中的变化趋势及其对EMT的调节作用，再通过基因敲除研究circ79530影响EMT进程和肾纤维化的机理，进一步利用小鼠模型和临床样本，探讨circ79530通过TWIST调节EMT的分子机制。本研究将揭示circ79530与TWIST形成正反馈回路调控肾纤维化EMT的作用，借助circRNA稳定易检出的特点，为肾纤维化的实时检测和有效控制提供新的诊疗策略，具有重要的战略意义和应用价值。

TWIST作为调控肾小管上皮细胞间充质转化（EMT）的核心转录因子，通过激活EMT促进肾纤维化引发肾衰竭。我们前期工作发现circRNA-79530(circ79530)通过与TWIST形成正反馈回路调节TWIST表达，其含量与EMT进程密切相关，提示circ79530是肾纤维化EMT过程中的重要调控因子。本项目通过体外培养HK-2和HKC，经细胞水平验证明确circRNA-79530可能通过抑制miR-214 发挥促进TWIST 表达的作用，通过进一步构建EMT模型，通过生物信息学分析，找出具有代表性的非编码RNA，经验证最终发现lncRNA-4474-miR-615-HNF-1β 能够形成一个调节链条，在肾纤维化过程中发挥重要的调节作用。因为基因序列的复杂性和多样性，lncRNA-4474 可能会与多种miRNA 相结合，miR-615 可能会有多个靶基因，并且HNF-1β 可能也会被多种miRNA 调节，而在肾纤维化发生发展的环境下，我们发现了一个新的调节链条，并且能够在肾纤维化过程中发挥促进或者抑制的作用。本项目在发现新机制的同时，也给肾纤维化检测及治疗提供了新的思路。