Identification of the non-proteolytic mechanism of NLRP1 activation

NLRP1 激活的非蛋白水解机制的鉴定

• 批准号: 9234453
• 负责人: Sarah E. Ewald
• 金额: $ 10.7万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9234453
• 关键词: Address; Alleles; Amino Acids; Anthrax disease; Area; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Award; Bacillus anthracis; Bacteria; Bacterial Infections; Binding; Biological Assay; Biology; Bone Marrow; Cell Death; Cells; Chemicals; Cleaved cell; Complement; Complex; Data; Detection; Development; Disease; Epitopes; Flagellin; Genes; Genetic Engineering; Genetic Screening; Goals; Human; Immune; Immune response; Immunofluorescence Immunologic; Immunology; Immunoprecipitation; Individual; Infection; Infectious Agent; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory Response; Innate Immune System; Intervention; K22 Award; Lead; Light; Mediating; Modeling; Molecular; N-terminal; Parasites; Pathway interactions; Peptide Hydrolases; Process; Production; Proteolysis; Rattus; Resources; Rest; Rodent; Role; Route; System; Time; Toxoplasma; Toxoplasma gondii; Training; Translating; Vaccines; Variant; Virus; Virus Diseases; Work; anthrax lethal factor; autoinflammatory; biochemical tools; cell injury; cell type; design; genetic approach; genome sequencing; immunosuppressed; in vivo; killings; macrophage; microbial; mutant; novel; pathogen; public health relevance; response; reverse genetics; sensor; skills; therapeutic development; therapy development; tool; tumor; vaccine development; vaccine response; vaccinology

 DESCRIPTION (provided by applicant): Inflammasomes are innate immune sensing pathways designed to identify and clear infectious agents through production of pro-inflammatory mediators and pyroptotic cell death. Selective targeting of inflammasomes and their sensors, the NLRPs, is emerging as an important means of modulating the immune response in vaccinology, tumor therapy and treatment of autoimmunity. However, only two molecular mechanisms of NLRP activation have been described to date: NLRP1 proteolysis by Bacillus anthracis lethal factor and flagellin binding by the NAIPs/NLRC4 inflammasome. I have recently determined that NLRP1 also drives a host-protective inflammasome response to the protozoan parasite Toxoplasma gondii. In contrast to the previously described mechanism of NLRP1 activation by anthrax, however, NLRP1 is not proteolytically processed in response to Toxoplasma infection, leading to the hypothesis that NLRP1 has evolved to detect parasite infection via a novel, undescribed mechanism. This K22 award will provide the experimental resources, time and training to identify the critical components of the NLRP1 sensing pathway. Specifically, this award will allow me to develop 1) skills in the design and implementation of forward genetic screens, 2) tools and expertise in reverse genetic engineering of parasites, 3) biochemical tools to probe the essential components of NLRP1 for sensor activation. These studies address an understudied area in immunology: how eukaryotic pathogens are sensed by the innate immune system. Moreover, by exploiting the natural allelic variation in NLRP1 in parallel with a parasite genetics approach I will establish a unique set of tools to define the molecular underpinnings of NLRP1 activation that will be broadly important to our understanding of inflammasome biology. Specifically, the central goals of this project are to 1) identify the parasite components that trigger NLRP1 activation and 2) determine the critical residues in NLRP1 that mediate this response.

 描述（由申请人提供）：炎性小体是先天性免疫传感途径，旨在通过产生促炎介质和致炎细胞死亡来识别和清除感染因子。选择性靶向炎性小体及其传感器，NLRP，正在成为疫苗学，肿瘤治疗和自身免疫治疗中调节免疫应答的重要手段。然而，迄今为止仅描述了两种NLRP激活的分子机制：炭疽杆菌致死因子引起的NLRP 1蛋白水解和NAIP/NLRC 4炎性体引起的鞭毛蛋白结合。我最近确定NLRP 1也驱动宿主对原生动物寄生虫弓形虫（Toxoplasma gondii）的保护性炎症反应。然而，与先前描述的炭疽激活NLRP 1的机制相反，NLRP 1在弓形虫感染时不被蛋白水解处理，这导致了NLRP 1已经进化为通过一种新的、未描述的机制来检测寄生虫感染的假设。该K22奖项将提供实验资源，时间和培训，以确定NLRP 1传感途径的关键组成部分。 具体来说，该奖项将使我能够发展1）正向遗传筛选的设计和实施技能，2）寄生虫反向遗传工程的工具和专业知识，3）探测NLRP 1传感器激活的基本组成部分的生化工具。这些研究解决了免疫学中一个未充分研究的领域：先天免疫系统如何感知真核病原体。此外，通过利用NLRP 1中的天然等位基因变异与寄生虫遗传学方法并行，我将建立一套独特的工具来定义NLRP 1激活的分子基础，这对我们理解炎性小体生物学非常重要。具体而言，该项目的中心目标是：1）确定触发NLRP 1激活的寄生虫组分; 2）确定NLRP 1中介导这种反应的关键残基。

项目成果

Immunoregulatory Role of NK Cells in Tissue Inflammation and Regeneration.

• DOI: 10.3389/fimmu.2017.00301
• 发表时间: 2017
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Tosello-Trampont A;Surette FA;Ewald SE;Hahn YS
• 通讯作者: Hahn YS