ROLE OF NEK7 PROTEIN IN NLRP3 INFLAMMASOME ACTIVATION AND INFLAMMATION

项目摘要/摘要 Nacht、LRR和PYD结构域包含蛋白3(NLRP3)属于节点样受体(NLR)的一类 触发炎症体组装的蛋白质，这是一个介导caspase-1的分子平台 激活、加工和分泌具有生物活性的IL-1β和IL-18。除了它在以下方面的关键作用 先天免疫，NLRP3炎症体的失调与遗传性和获得性有关 炎症性疾病，如低温相关自炎综合征、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在医学上具有重要意义 炎症体，其被激活的机制仍然不清楚。使用蛋白质组学方法揭示 在巨噬细胞中与NLRP3相互作用的关键因素，我们确定Nek7蛋白激酶是一种NLRP3- NLRP3炎症体组装所必需的相互作用蛋白。Nek7与 NLRP3处于静息状态，这种相互作用在NLRP3激活过程中得到加强。在巨噬细胞中 Caspase-1的激活和IL-1的分泌被Nek7耗尽，以响应触发的刺激 NLRP3激活。相反，Nek7不是激活NLRC4和AIM2炎症体所必需的。 Nek7在激活NLRP3炎症小体中的关键和特定作用是意想不到的，因为 Nek7是调节细胞质分裂的激酶家族的成员。这项提案的目标是定义 Nek7调节NLRP3炎症体激活的分子机制。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 通过促进炎性小体的组装来激活。此外，我们假设Nek7 导致炎症性疾病的诱发和/或进展。为了检验这些假设，我们建议 Nek7调控NLRP3炎性小体激活的机制及其作用 在体内使用依赖于通过NLRP3分泌IL-1的动物炎症模型。了解企业的角色 NLRP3炎症体激活中的Nek7有望为小说的发展提供关键的见解 炎症性疾病的治疗策略。

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.