mTOR信号通路异常激活已被证实参与颞叶内侧癫痫发病过程，目前研究发现mTOR上游负调控蛋白DEPDC5突变激活mTOR通路诱导癫痫发生，但其机制尚不明确。项目组前期研究发现mTOR表达异常升高的颞叶内侧癫痫患者中DEPDC5编码基因存在3个重复发生的突变，体外结构模型分析显示其中一个未见报道的错义突变p.Trp1242Ala位于DEP功能结构域，初步证实DEPDC5突变的致痫作用。然而DEPDC5基因突变如何影响DEPDC5蛋白功能，进而对mTOR通路产生失抑制作用尚不明确。因此，本项目拟利用颞叶内侧癫痫患者手术切除海马组织，研究DEPDC5突变对DEPDC5表达水平与mTOR信号通路活性的作用，并在体外表达纯化全长DEPDC5等蛋白，基于蛋白质相互作用及蛋白结构，探索突变DEPDC5蛋白参与mTOR活化及颞叶内侧癫痫的病理机制，为阐明颞叶内侧癫痫发病机制及寻找治疗新靶点提供新思路。

Aberrant activation of mTOR signaling pathway has been confirmed to be involved in the pathogenesis of mesial temporal lobe epilepsy. Recently, it has been found that mutation of DEPDC5, a negative regulator of mTOR, induce epilepsy by activating the mTOR pathway, but its mechanism remains unclear. In the previous study of our project, three rare variants of the DEPDC5 gene were found in patients with abnormally elevated mTOR expression in temporal lobe epilepsy. In silico structural model analysis revealed that one of the unreported missense mutations p.Trp1242Ala was located in the DEP functional domain，which preliminary confirmed its epileptogenic effect. However, how the DEPDC5 mutation affects the function of DEPDC5 protein, and the further dysregelation effect of mTOR pathway is not clear. Therefore, our project intends to use the surgical resection of hippocampal tissue in patients with temporal lobe epilepsy to study the effect of DEPDC5 mutation on DEPDC5 expression and mTOR signaling pathway activity, and further express and purify full-length DEPDC5 and other proteins in vitro, by means of protein interaction and structure, to explore the mechanism of mutated DEPDC5 protein in the pathogenesis of mTOR activation and medial temporal lobe epilepsy, for providing new ideas of elucidating the pathogenesis of temporal lobe epilepsy and finding new therapeutic targets.