METTL14介导的m6A甲基化修饰调控mTOR通路对角膜移植免疫排斥的影响及机制研究

免疫排斥是角膜移植失败的最主要原因。本团队的既往研究发现：抑制mTOR通路活性能有效防治角膜移植排斥反应，但防控新思路如发挥更大的作用，则需明确mTOR上游的关键调控因子及机制。前期预实验显示：角膜移植排斥过程中m6A甲基转移酶METTL14表达显著下降，而过表达的METTL14通过抑制mTOR通路活性缓解角膜移植免疫排斥反应。因此，我们提出“METTL14介导的m6A甲基化修饰通过负调控mTOR通路活性抑制角膜移植排斥”的假说。为验证此假说，本项目拟：（1）研究METTL14对小鼠角膜移植免疫排斥和mTOR通路活性的影响；（2）阐明METTL14调控mTOR通路活性的关键机制；（3）利用METTL14转基因小鼠验证其对角膜移植免疫排斥以及mTOR通路活性的影响。通过本研究初步阐明m6A修饰调控角膜移植mTOR通路的关键机制，为临床高效防治角膜移植免疫排斥反应提供理论基础。

Corneal allograft rejection was the main cause for transplant failure. Our group’s previous research revealed that increased autophagy through blocking mTOR signaling pathway significantly alleviated corneal allograft rejection. However, to maximize the role of this new approach to prevention and treatment of allograft rejection, further studies are needed to investigate how mTOR signaling pathway is regulated during corneal allograft rejection. Our results showed that the less expression of m6A methyltransferase METTL14 in rejected corneal allografts, while overexpression of METTL14 pronouncedly allerviated corneal allograft rejection through blockade of mTOR activity. Based on this, we hypothesized that METTLL14 alleviated corneal allograft rejection via inhibiting mTOR signaling pathway by m6A methylation modification. To test this hypothesis, we will intend to (1) investigate the effect of METTL14 on corneal allograft rejection and mTOR signaling pathway using murine corneal transplantation model; (2) elucidate the mechanism of METTL14 played on mTOR signaling pathway; (3) verify the effect of METTL14 on corneal allograft rejection and mTOR signaling pathway using METTL14 transgenic mice. This study will clarify the effect and mechanisms of m6A methylation modification mediated by METTL14 on corneal allograft rejection and mTOR signaling, and provide a theoretical basis clinically effective prevention and treatment of corneal allograft rejection.