基于ATAC-seq技术研究交叉反应物质197调控TFEB介导的自噬抑制子宫内膜异位症侵袭的分子机制

内异症的治疗存在瓶颈问题。我们的前期研究发现，交叉反应物质197（CRM197）可有效抑制内异症的侵袭，但作用机制不清。鉴于内异症细胞在内异症微环境中受到多种复杂调控，本项目基于ATAC-seq从染色质可接近性水平探索CRM197的调控机制。预实验结果发现，CRM197可有效增强自噬转录因子TFEB的核转位和细胞自噬活性；siRNA沉默TFEB则可显著减弱CRM197增强自噬和抑制内异症侵袭的能力。鉴此，我们提出假说：CRM197通过调节染色质可接近性增强TFEB介导的自噬，从而抑制内异症侵袭。为验证这一假说，拟在细胞、裸鼠、人体水平，从信号转导途径、表观遗传学及调控机理的层面，采用WB、IF、CHIP-qPCR、RNA干扰等技术，从“染色质可接近性-TFEB-自噬”的新视角明确CRM197抑制内异症侵袭的作用与机制。本项目将有望为内异症的治疗提供安全有效的新药物和新靶点。

There is a bottleneck problem in the treatment of endometriosis. Our previous study found that the Cross-reacting material 197 (CRM197) effectively inhibit the invasion of ectopic endometrial cells, but the mechanism is unclear. Considering that the endometrial cells are regulated by a variety of complex regulation in the endometriosis microenvironment, this study explored the regulatory mechanism of CRM197 based on ATAC-seq at the chromatin accessibility level. The pre-experiments in this study suggested that CRM197 could significantly enhance the activity of autophagy transcription factor TFEB and autophagy; siRNA silencing of TFEB significantly attenuated CRM197's ability to enhance autophagy and inhibit the invasion of endometriosis. Therefore, we hypothesized that CRM197 might inhibit the invasion of endometriosis via enhancing TFEB-mediated autophagy by regulating chromatin accessibility. To prove this hypothesis, we will elucidate the mechanism of the effect and mechanism of CRM197 on inhibiting endometriosis invasion from the new point of chromatin accessibility-TFEB-autophagy and the aspects of signal transduction pathway, epigenetics and regulatory mechanism, using WB, immunofluorescence staining, confocal microscopy, CHIP-qPCR, RNA interference, transfection and other techniques at the cell, nude mouse and human levels. This project will provide safe and effective new drugs and targets for endometriosis treatment.