靶向NLRP3炎症小体的小分子抗炎药物筛选及分子药理机制研究

NLRP3 inflammasome plays a critical role in innate immunity and inflammation，and is involved in the development of several major human diseases, including type 2 diabetes, neurodegenerative diseases and tumor, but the detailed mechanisms of NLRP3 inflammasome activation and regulation and the anti-inflammatory strategy targeting NLRP3 inflammasome remain little known. We recently found that ω-3 fatty acids could prevent high fat diet-induced metabolic disorder by inhibition of NLRP3 inflammasome (Immunity, In press), suggesting the possibility to prevent the progression of inflammation-associated diseases by inhibition of NLRP3 inflammasome. In this project, we will screen the inhibitors of NLRP3 inflammasome using low-molecular-weight compound libraries, and then will identify the critical proteins or signal pathways involved in NLRP3 inflammasome activation by analyzing the role of the targets of the inhibitors. After that, we will further study the mechanisms of how these proteins or signal pathways regulate NLRP3 inflammasome activation. Based on these results, we will investigate the effects of these NLRP3 inflammasome inhibitors on high fat diet-induced type 2 diabetes model and LPS-induced sepsis model. This project not only is helpful for clarification of the mechanisms of NLRP3 inflammasome activation and regulation, but also is helpful for identification new anti-inflammatory compounds targeting for NLRP3 inflammasome and providing experimental evidences for therapy of inflammation-associated diseases.

NLRP3炎症小体在固有免疫应答和炎症发生中起关键作用且与2型糖尿病、神经退行性疾病和肿瘤等多种人类重大疾病密切相关，但目前对其活化和调控机制知之甚少，也缺乏靶向NLRP3炎症小体的抗炎干预策略。我们近期发现ω-3脂肪酸分子能通过抑制NLRP3炎症小体活化缓解高脂食物诱导的代谢紊乱（Immunity，In press），提示可以靶向NLRP3炎症小体干预炎症相关疾病进程。本课题拟利用小分子药物文库筛选NLRP3炎症小体的抑制剂，并以此为基础鉴定NLRP3炎症小体活化和调控中的关键分子和信号通路，并阐明其调控机制；进而利用高脂食物诱导的2型糖尿病模型和LPS诱导的脓毒血症模型，探究筛选所得靶向NLRP3炎症小体抗炎药物对疾病的干预效果。该项目的实施不仅有助于阐明NLRP3炎症小体的活化和调控机制，也有助于寻找靶向NLRP3炎症小体的抗炎药物，并为炎症相关重大疾病治疗提供实验依据。

NLRP3炎症小体在固有免疫应答和炎症发生中起关键作用且与2型糖尿病、神经退行性疾病和肿瘤等多种人类重大疾病密切相关，但目前对其活化和调控机制及靶向抗炎干预策略还知之甚少。我们前期发现w-3脂肪酸分子能通过抑制NLRP3炎症小体活化缓解高脂食物诱导的代谢紊乱（Immunity，2013），提示NLRP3炎症小体是潜在的炎症相关疾病干预靶点。本课题拟利用小分子药物文库筛选NLRP3炎症小体的抑制剂，探究药物靶点对NLRP3炎症小体活化的影响，鉴定NLRP3炎症小体活化和调控中的关键分子和信号通路，并阐明其调控机制；进而利用动物模型，探究靶向NLRP3炎症小体的抗炎药物对疾病的干预效果。本项目主要取得如下研究成绩：1）已经完成针对NLRP3的10000个小分子药物文库筛选，得到20余个可正向或者负向调控NLRP3炎症小体活化的小分子化合物； 2）阐明了小分子多巴胺，Necrostatin-1，CY-09、Oridonin和IAA94抑制NLRP3炎症小体的分子机制；3）鉴定了DRD1-cAMP,RIP-1RIP3, CLICs等调控NLRP3炎症小体活化的信号通路或者分子；4）发现CY-09和Oridonin可直接靶向NLRP3抑制NLRP3炎症小体活化并对NLRP3驱动的疾病，比如2型糖尿病和痛风等疾病具有较好的预防和治疗效果。5）以通讯作者发表课题编号标注的论文6篇，包括Cell、Nat Immunol、J Exp Med、Nat Commu(2)、Trends Immunol，授权专利1项。该项目的实施不仅鉴定了NLRP3炎症小体重要的活化和调控信号通路，也有助于发展靶向NLRP3的疾病干预药物。