Setd7靶向p21调控角膜上皮损伤修复的分子机制研究

角膜上皮的损伤修复对于维持角膜的完整性至关重要，其分子调控机制尚未明了。组蛋白甲基化修饰是一种极为重要的表观遗传调控机制，我们的前期工作发现：角膜上皮损伤修复中组蛋白甲基化转移酶Setd7以及H3K4me下调，同时p21也明显降低，提示Setd7介导的H3K4me靶向p21调控角膜上皮的损伤修复。在此基础上，本项目首先联合采用质谱和生物信息学等技术，系统分析小鼠角膜损伤修复模型中组蛋白甲基化修饰图谱。进而以人角膜上皮细胞为模型，通过基因干扰和过表达技术，明确Setd7的生物学功能。通过ChIP明确Setd7 靶向p21的表观调控机制以及H3K4me修饰的靶位点。最后通过小鼠模型在体验证Setd7对角膜上皮的损伤修复的效应机制。本研究将阐明Setd7介导的组蛋白甲基化在角膜上皮损伤修复中的分子调控机制，为角膜损伤相关疾病的临床治疗提供新的思路。

Corneal epithelial wound healing (CEWH) plays a critical role in maintaining corneal integrity, but the mechanisms involved in this process remain to be investigated in depth. Histone methylation modifications are important components of epigenetic regulation. We performed preliminary studies showing that during wound healing there are significant changes occurring in H3K4 methyltransferase Setd7, H3K4me and p21 expression during this process. Such changes suggest that Setd7-mediated H3K4me targeted p21 can be important effectors of the CEWH process. On the basis, we first established a mice CEWH model to extract epithelial histones. Two dimensional-nano-LC-MS/MS analysis was performed along with western blot and real time qRT-PCR to identify the differences in histone methylation expression patterns between normal control epithelial cells and those involved in the wound healing process. Human corneal epithelial cell specific for Setd7 losses or gains of function were generated to determine their effects on these epithelial cellular responses to injury. Additionally, molecular mechanisms of Setd7-targeted p21 will be identified by ChIP. The functional mechanisms of specific Setd7 on CEWH will also be verified in an in vivo model. This study will clarify the molecular mechanisms of Setd7-mediating control of H3K4 methylation status during CEWH as well as identify novel potential drug targets for more selective intervention and in treatment of patients with corneal wounds compromising the CEWH process.