ROLE OF NEK7 PROTEIN IN NLRP3 INFLAMMASOME ACTIVATION AND INFLAMMATION

NEK7 蛋白在 NLRP3 炎症小体激活和炎症中的作用

• 批准号: 9179168
• 负责人: Yuan He
• 金额: $ 16.04万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-17 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9179168
• 关键词: Alzheimer' s Disease; Animal Model; Atherosclerosis; Attenuated; Biological; CASP1 gene; Co-Immunoprecipitations; Crohn' s disease; Crystallization; Cyclic AMP-Dependent Protein Kinases; Cytokinesis; Development; Diabetes Mellitus; Disease; Event; Family; Family member; Goals; Gout; Inflammasome; Inflammation; Inflammatory; Inherited; Interleukin-1; Interleukin-1 beta; Interleukin-18; Knock-in Mouse; Lead; Link; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Medical; Modeling; Molecular; Mus; Mutation; Natural Immunity; Pathogenesis; Periodicity; Peritoneal; Phosphotransferases; Play; Protein Kinase; Proteins; Proteomics; Rest; Role; Signal Pathway; Signal Transduction; Stimulus; Syndrome; Testing; Urate; autoinflammatory; base; in vivo; insight; macrophage; member; mouse model; mutant; novel therapeutics; protein complex; receptor; response

Project Summary/Abstract NACHT, LRR and PYD domains-containing protein 3 (NLRP3) belongs to a class of Nod-like-receptor (NLR) proteins that trigger the assembly of the inflammasome, a molecular platform that mediates caspase-1 activation and processing and secretion of biologically active IL-1β and IL-18. In addition to its critical role in innate immunity, dysregulation of the NLRP3 inflammasome has been linked to both inherited and acquired inflammatory disorders, such as Cryopyrin-associated autoinflammatory syndrome, gout, Crohn’s disease, Alzheimer’s disease, diabetes and atherosclerosis. Despite the medical importance of the NLRP3 inflammasome, the mechanism by which it is activated remains elusive. Using a proteomics approach to reveal critical factors that interact with NLRP3 in macrophages, we identified the Nek7 protein kinase as a NLRP3- interacting protein that is essential for the assembly of the NLRP3 inflammasome. Nek7 is associated with NLRP3 in the resting state, and this interaction is enhanced during NLRP3 activation. In macrophages depleted with Nek7, caspase-1 activation and IL-1 secretion are abrogated in response to stimuli that trigger NLRP3 activation. In contrast, Nek7 is not required for the activation of the NLRC4 and AIM2 inflammasomes. The critical and specific role of Nek7 in the activation of the NLRP3 inflammasome is unexpected because Nek7 is a member of a kinase family that regulates cytokinesis. The goal of this proposal is to define the molecular mechanism by which Nek7 regulates the activation of the NLRP3 inflammasome. Based on our preliminary results, our primary hypothesis is that the Nek7 protein kinase regulates NLRP3 inflammasome activation by promoting the assembly of the inflammasome. Furthermore, we hypothesize that Nek7 contributes to the induction and/or progression of inflammatory disease. To test these hypotheses, we propose characterizing the mechanism by which Nek7 regulates NLRP3 inflammasome activation and the role of Nek7 in vivo using animals models of inflammation that rely on IL-1 secretion via NLRP3. Understanding the role of Nek7 in NLRP3 inflammasome activation is expected to provide critical insight into the development of novel therapeutic strategies for inflammatory diseases.

项目概要/摘要 含有 NACHT、LRR 和 PYD 结构域的蛋白 3 (NLRP3) 属于一类 Nod 样受体 (NLR) 触发炎症小体组装的蛋白质，炎症小体是介导 caspase-1 的分子平台 具有生物活性的 IL-1β 和 IL-18 的激活、加工和分泌。除了它的关键作用 先天免疫，NLRP3 炎症小体的失调与遗传性和获得性有关 炎症性疾病，例如冷热蛋白相关的自身炎症综合征、痛风、克罗恩病、 阿尔茨海默病、糖尿病和动脉粥样硬化。尽管 NLRP3 具有医学重要性 炎症小体，其激活机制仍然难以捉摸。使用蛋白质组学方法来揭示 与巨噬细胞中 NLRP3 相互作用的关键因素，我们将 Nek7 蛋白激酶鉴定为 NLRP3- NLRP3 炎症小体组装所必需的相互作用蛋白。 Nek7 关联于 NLRP3 处于静息状态，并且这种相互作用在 NLRP3 激活期间增强。在巨噬细胞中 Nek7 耗尽后，caspase-1 激活和 IL-1 分泌会响应触发的刺激而被消除 NLRP3 激活。相比之下，NLRC4 和 AIM2 炎症小体的激活不需要 Nek7。 Nek7 在 NLRP3 炎症小体激活中的关键和特定作用是出乎意料的，因为 Nek7 是调节胞质分裂的激酶家族的成员。该提案的目标是定义 Nek7 调节 NLRP3 炎症小体激活的分子机制。基于我们的 初步结果，我们的主要假设是 Nek7 蛋白激酶调节 NLRP3 炎症小体 通过促进炎症小体的组装来激活。此外，我们假设 Nek7 有助于炎症性疾病的诱导和/或进展。为了检验这些假设，我们建议 表征 Nek7 调节 NLRP3 炎性体激活的机制以及 Nek7 的作用 使用依赖于通过 NLRP3 分泌 IL-1 的炎症动物模型进行体内实验。了解角色 NLRP3 炎症小体激活中的 Nek7 有望为新型药物的开发提供重要见解 炎症性疾病的治疗策略。