VRAC通道调控NLRP3炎症小体活化的发现与机制及其作为多发性硬化症治疗新靶点的研究

过度活化的NLRP3炎症小体是多发性硬化症等神经免疫疾病的重要病理诱因。氯离子外流和细胞肿胀直接活化NLRP3炎症小体，然而，机制尚不清楚。体积调控型阴离子通道VRAC是一类重要的渗透压敏感型离子通道，LRRC8家族蛋白在2014年被报道介导VRAC通道，LRRC8A需与LRRC8B-E异聚形成VRAC通道。我们已针对LRRC8开展了大量结构-功能学和生理学研究，发现巨噬细胞和小胶质细胞中表达显著VRAC电流，沉默LRRC8A可显著抑制该电流。VRAC抑制剂DCPIB或沉默LRRC8A均可抑制NLRP3炎症小体活化。基于上述研究基础，本项目将综合采用电生理学、分子细胞生物学和基因敲除小鼠等方法研究VRAC通道对NLRP3炎症小体活化的调控及机制，并进一步探索LRRC8A对多发性硬化症等神经免疫疾病的影响，为相关疾病的发病机制和药物新靶点的发现提供理论依据。

NLRP3 inflammasome plays a critical role in neuroinflammation diseases such as multiple sclerosis. Recent studies have shown that Cl- efflux and cell swelling could directly activate NLRP3 inflammasome, however, the mechanism is still unclear. Volume-regulated anion channels (VRAC) belong to osmolarity sensitive channels and the underlying molecular determinants are LRRC8 proteins, which was revealed in 2014. LRRC8A is an essential component of VRAC, while LRRC8A must co-assemble with at least one member of LRRC8B-E to form functional channels. We have carried out structural-functional and physiological studies on LRRC8 proteins and found that inhibiting VRAC channels or silencing LRRC8A significantly inhibited the activation of NLRP3 inflammasome. Based on above results, this project will comprehensively utilize electrophysiology, molecular cell biology, and knockout mice to study the regulation and mechanism of VRAC channels on NLRP3 inflammasome activation and explore the role of LRRC8A in multiple sclerosis. Our project will provide a theoretical basis for the pathogenesis and novel drug targets identification of neuroinflammation related diseases.