A Novel CDN Sensor-Adaptor System for Host Defense Against Infection

用于主机防御感染的新型 CDN 传感器适配器系统

基本信息

批准号：
10165463
负责人：
John David MacMicking
金额：
$ 41.88万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-01 至 2022-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10165463
关键词：
Acute Adaptor Signaling Protein Affinity Anti-Bacterial Agents Antibiotic Therapy Autophagocytosis Bacteria Binding Binding Proteins Biological Process Blood Bypass Cause of Death Cells Chimera organism Cytosol Data Detection Dinucleoside Phosphates Engineering Enzymes Event Family Family member GBP1 gene Gastroenteritis Guanosine Triphosphate Phosphohydrolases Host Defense Human Immune Immune response Immunity In Vitro Infection Inflammasome Intensive Care Units Interferon Type I Interferons Ligands Link Listeria monocytogenes Mammals Mediating Modeling Molecular Mus Mycobacterium tuberculosis Nucleotides Organism Periodicity Physiological Population Production Proteins Publishing Salmonella typhimurium Second Messenger Systems Sepsis Signal Transduction Stimulator of Interferon Genes Structure System Testing Tuberculosis antimicrobial base defense response design ds-DNA experimental study fighting foodborne genetic approach guanylate human disease in vivo member microbial microbial signature mutant novel paracrine pathogen pathogenic bacteria pathogenic microbe programs protein function public health relevance recruit response sensor

项目摘要

PROJECT SUMMARY/ABSTRACT Mammals defend against infection by detecting foreign and/or danger signals to mount both cell-autonomous as well as paracrine immune responses. Among the microbial signatures sensed by mammalian hosts are a new class of signaling intermediates termed cyclic di-nucleotides (CDNs). CDNs are recognized by the host protein, STING (STimulator of INterferon Genes), to elicit type I IFN production. In this application we will characterize a new CDN recognition system that mobilizes protective immunity independently of STING or type I IFN release. This new circuit enlists four members of an immune GTPase family – the 65-73kDa Guanylate Binding Proteins (GBPs) – to activate distinct host defense programs in both humans and mice. Preliminary evidence implicates GBP1 as the proximal CDN sensor which recruits GBP2, GBP3 and GBP5 as adaptors that link the antimicrobial machinery further downstream. In Aim 1, we will test the importance of GBP1 to act as a broad-spectrum CDN sensor capable of binding c-diGMP, c-diAMP and cGAMPs under physiological conditions to elicit either inflammasome assembly, pyroptosis or autophagy in a STING-independent manner. This will be examined using a powerful host-pathogen genetic strategy where mice and cells lacking GBP1 or expressing GBP1 CDN-binding mutants will be infected with important bacterial pathogens (M. tuberculosis, S. typhimurium and L. monocytogenes) engineered to secrete a specific CDN moiety to define this relationship in vitro and in vivo. In Aim 2, we expand this CDN sensor circuit to include GBP2, GBP3 and GBP5 as heterotypic partners that direct core components of the inflammasome, pyroptotic or autophagy cascade to restrict bacterial replication. This aim will enlist newly-created Gbp2-/-, Gbp3-/- and Gbp5-/- mice as well as mixed sensor-adaptor chimeras as a novel binary approach to dissect how the GBP relay mobilizes different types of CDN-dependent immunity to bacterial pathogens. Collectively, our proposal examines a new set of host proteins that define a novel CDN sensor-adaptor circuit with important implications for developing host- directed therapies as an adjunct to standard antibiotic treatment.

项目概要/摘要哺乳动物通过检测外来和/或危险信号来防御感染，以安装细胞自主的以及旁分泌免疫反应是哺乳动物宿主感知到的微生物特征之一。称为环状二核苷酸（CDN）的新型信号中间体可以被宿主识别。蛋白质，STING（干扰素基因的 STimulator），用于引发 I 型干扰素的产生。描述了一种新的 CDN 识别系统，该系统可以独立于 STING 或类型来调动保护性免疫 I IFN 释放。这个新电路招募了免疫 GTP 酶家族的四个成员 - 65-73kDa 鸟苷酸。结合蛋白 (GBP) – 激活人类和小鼠的不同宿主防御程序。有证据表明 GBP1 是近端 CDN 传感器，它招募 GBP2、GBP3 和 GBP5 作为适配器在目标 1 中，我们将测试 GBP1 发挥作用的重要性。作为广谱 CDN 传感器，能够在生理条件下结合 c-diGMP、c-diAMP 和 cGAMP 以不依赖于 STING 的方式引发炎症小体组装、细胞焦亡或自噬的条件。这将使用强大的宿主病原体遗传策略进行检查，其中小鼠和细胞缺乏 GBP1 或表达 GBP1 CDN 结合突变体将被重要的细菌病原体（结核分枝杆菌、链球菌）感染。鼠伤寒杆菌和单核细胞增生利斯特氏菌）被设计为分泌特定的 CDN 部分来定义这种关系在目标 2 中，我们扩展了该 CDN 传感器电路以包括 GBP2、GBP3 和 GBP5。异型伙伴将炎症小体、焦亡或自噬级联的核心成分引导至该目标将招募新创建的 Gbp2-/-、Gbp3-/- 和 Gbp5-/- 小鼠以及混合传感器适配器嵌合体作为一种新颖的二元方法来剖析 GBP 继电器如何调动不同的总的来说，我们的提案研究了一组新的 CDN 依赖性免疫。宿主蛋白定义了一种新型 CDN 传感器适配器电路，对开发宿主具有重要意义定向治疗作为标准抗生素治疗的辅助手段。