Trim27调控PPARγ泛素化在获得性癫痫发生中的作用及机制

获得性癫痫（AE）是目前已知存在形成风险但又缺乏有效预防手段仅有的几种脑部疾病之一。核转录因子PPARγ可通过平衡神经炎症、调控小胶质细胞表型和功能在阻抑AE形成中起关键作用, 申请人证实PPARγ促进癫痫持续状态后小胶质细胞从促炎M1向抑炎M2型转化，并发挥神经保护作用。项目组前期发现癫痫患者颞叶组织的PPARγ明显减少，E3泛素连接酶Trim27高表达，且与PPARγ的表达负相关；在谷氨酸损伤中Trim27泛素化降解PPARγ，但Trim27泛素化调控PPARγ在AE形成中的作用及机制尚不明确。本项目拟采用过表达/沉默策略，结合细胞、动物模型，论证Trim27在AE形成中的作用；利用体内外生化技术，研究Trim27泛素化调控PPARγ的分子机制；使用转录组测序和基因敲除技术，探讨Trim27泛素化调控PPARγ对下游AE形成信号的作用途径和靶基因，为预防AE提供新的作用靶点和治疗手段。

Acquired epilepsy (AE) is one of the few brain diseases known to have risk of development but lack of effective prevention. Imbalance of neuroinflammation, abnormal function and phenotype of microglia promote the development of AE. Nuclear transcription factor, PPARγ, plays a key role in inhibiting the development of AE by balancing neuroinflammation and regulating the phenotype and function of microglia. In the preliminary study of our project group, it was found that PPARγ was significantly decreased, while E3 ubiquitin ligase, Trim27, was highly expressed in temporal lobe tissue of patients with epilepsy with negatively correlation with the expression of PPARγ. Trim27 Ubiquitinates and degrades PPARγ in glutamate injury, but the role and mechanism of Trim27 ubiquitination regulation of PPARγ in AE develepment is not clear. This project intends to adopt overexpression/silencing strategy, combined with cellular and animal models, to determine the role of Trim27 in AE development; to study the molecular mechanism of Trim27 ubiquitination regulation of PPARγ by in vivo and in vitro biochemical techniques; and to explore the pathway and target genes of Trim27 ubiquitination regulation of PPARγ on downstream AE development signals and target genes by transcriptome sequencing and gene knockout techniques, so as to provide new targets and therapeutic means for the prevention of AE development.