Regulation of LPS-responses by ZBP1

ZBP1 对 LPS 反应的调节

• 批准号: 10808282
• 负责人: Alexander Poltorak
• 金额: $ 23.42万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10808282
• 关键词: Acute; Address; Affect; Bacterial Infections; CASP8 gene; CD14 gene; Cell Death; Cell Death Induction; Cessation of life; Complex; Cytosol; Data; Endosomes; Equilibrium; Experimental Models; Gene Silencing; Goals; Gram-Negative Bacteria; Gram-Negative Bacterial Infections; Hypotension; Immune response; Immunoprecipitation; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Intervention; Investigation; Kinetics; Knock-out; Ligation; Lipopolysaccharides; MAP Kinase Gene; Macrophage; Mediating; Mediator; Modeling; Multiple Organ Failure; Mus; Nitric Oxide; Nucleic Acids; Outcome; Pathway interactions; Phosphotransferases; Physiological; Play; Poison; Positioning Attribute; Predisposition; Production; Protein Kinase Interaction; Publications; Publishing; RIPK1 gene; Regulation; Reporting; Research; Resistance; Role; Scaffolding Protein; Secure; Sepsis; Septic Shock; Shock; Signal Transduction; Stress; Surface; TLR3 gene; TLR4 gene; TNFRSF1A gene; Tissues; Viral; Virus Diseases; Yersinia; Yersinia infections; cytokine; in vivo; innate immune pathways; insight; member; monocyte; neutrophil; new therapeutic target; novel; pathogen; pharmacologic; recruit; response; sensor; synergism; trafficking

PROJECT SUMMARY The goal of this proposal is investigation of the mechanism of ZBP1-mediated response to LPS and protection from bacterial infection that is provided by ZBP1. Our published and preliminary data characterize a novel noncanonical activation fo Caspase 8 in response to LPS, which results in the pyroptotic cell death in macrophages. In further inquiry, we identified that containing CASP8 pro-death complex II that assembles in response to LPS, is not dependent on TNFR1 as was widely accepted so far, but rather on TRIF thus making TRIF a central component of theis activation pathway. To reflect the requirement for TRIF in complex II formation and activation of CASP8, we termed this new pyroptosis-inducing complex the “TRIFosome”. In further mechanistic inquiry, we found that the nucleic acid sensor, ZBP1, which thus far has been implicated only in viral infections, plays a significant role in the formation of the TRIFosome complex by shuttling RIPK1 to TRIF. The reliance of Complex II formation on ZBP1 raised a question whether ZBP1 mediates inflammatory responses to LPS as well. In support of this hypothesis, our preliminary data show that ZBP1 mediates LPS- induced inflammatory responses, thus providing rationale for further in vivo investigations of the role that ZBP1 might play in responses to LPS and Yersinia infection. To address whether ZBP1-mediated response to LPS provides protection from the bacterial infection, we propose 2 aims. In Aim 1, we will investigate ZBP1- mediated in vitro response to LPS using double knockout and gene-silencing approach. We will use ZBP1- specific immunoprecipitation in order to determine the components of ZBP1-interactome. determine the contribution of ZBP1 to LPS-induced immune response, which will include elucidation specific role of ZBP1 in mediating TRIF-dependent and TRIF-independent responses. It will also include delineation of the role of ZBP1 in the RIPK1-specific response to LPS. including understanding the roles of the functional domains of ZBP1 in this pathway, analysis of the recruitment of ZBP1 into the endosomal TRIFosome complex, and characterization of the ZBP1-dependent changes in host response to bacterial infection in vivo. In the second Aim, we will capitalize on our preliminary findings and will further investigate how the kinetics of response to LSP that is conferred by ZBP1 confers sensitivity to LPS-induced tosicity I vivo, and whether this sensitivity provides protection from gram-negative infection in vivo. These studies will help establishing the role of ZBP1 in host response to bacterial infection and will position ZBP1 at the crosstalk of different immune response pathways.

项目摘要 该提案的目的是调查ZBP1介导的对LP和保护的反应机制 来自ZBP1提供的细菌感染。我们出版的和初步的数据是一个小说的特征 响应LPS的非规范激活FO caspase 8，这导致凋亡细胞死亡中的死亡 巨噬细胞。在进一步的询问中，我们确定包含CASP8 Pro-Death Complex II的组装 对LP的反应，不依赖于TNFR1，因为到目前为止被广泛接受，而是对TRIF的依赖 trif是theis激活途径的中心分量。以反映复合物II中TRIF的要求 CASP8的形成和激活，我们称这种新的凋亡诱导的复合物为“三斑点”。 进一步的机械询问，我们发现核酸传感器ZBP1迄今已涉及 仅在病毒感染中，通过将Ripk1的将RIPK1穿梭为 trif。 ZBP1对复合物II形成的依赖提出了一个问题，ZBP1是否介导炎症 对LP的回应。为了支持这一假设，我们的初步数据表明，ZBP1介导了LPS- 诱导炎症反应，从而为进一步研究ZBP1的作用提供了理由。 可能会对LP和Yersinia感染做出反应。解决ZBP1介导的对LPS的反应 我们提出了2个目标，可保护细菌感染。在AIM 1中，我们将研究ZBP1- 使用双基因敲除和基因分解方法介导了对LPS的体外反应。我们将使用ZBP1- 特定的免疫沉淀以确定ZBP1相互肌的成分。确定 ZBP1对LPS诱导的免疫响应的贡献，其中包括阐明ZBP1在 介导TRIF依赖性和与TRIF无关的响应。它还将包括划定的作用 RIPK1对LPS的特异性响应中的ZBP1。包括了解的功能领域的作用 ZBP1在此途径中，将ZBP1募集到内体三斑点复合体中的分析和 ZBP1依赖性变化在体内对细菌感染的反应的表征。在第二个 目的，我们将利用我们的初步发现，并将进一步调查如何回应的动力学 由ZBP1赋予的LSP承认对LPS诱导的Tosicition I Vivo的敏感性以及该灵敏度是否 提供免受体内革兰氏阴性感染的保护。这些研究将有助于确定ZBP1的作用 在宿主对细菌感染的反应中，将在不同免疫反应的串扰处定​​位ZBP1 途径。