Identification of Mycobacterium tuberculosis genes that mediate inhibition of the host cell IFN-beta signaling

介导宿主细胞 IFN-β 信号传导抑制的结核分枝杆菌基因的鉴定

基本信息

批准号：
9901025
负责人：
VOLKER BRIKEN
金额：
$ 22.78万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2021-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9901025
关键词：
Apoptotic Bacteria Cell Death Cell Line Cells Cessation of life Complement Cosmids DNA Data Dendritic Cells Detection Future Genes Genetic Screening Genome Growth IRF3 gene Immune response In Vitro Individual Infection Inflammasome Interferon-alpha Interferon-beta Interferons Knowledge Lead Libraries Luciferases M. tuberculosis genome Measures Mediating Modeling Molecular Molecular Analysis Mus Mycobacterium smegmatis Mycobacterium tuberculosis Pathway interactions Phenotype Population Production Proteins Public Health Publishing Reporter Reporter Genes Research Resources Series Signal Transduction Site Testing Time Tuberculosis Type I Epithelial Receptor Cell Virulence autocrine base drug development gain of function improved in vivo interest loss of function macrophage mutant new therapeutic target novel paracrine pathogen receptor screening tuberculosis drugs

项目摘要

Abstract It is estimated that about one third of the world’s population is latently infected with Mycobacterium tuberculosis (Mtb). The 10 million annual new cases of tuberculosis resulting in about one million deaths further underline the global public health impact of this pathogen. Mtb has developed a multitude of pathways to evade the host immune response. For the most part, the molecular mechanisms of these host pathogen interactions are only beginning to be understood. The premise of the current proposal is supported by a series of novel and unpublished results which demonstrate that Mtb inhibits IFN-a/b-receptor (IFNAR)-mediated cell signaling which results in improved intracellular growth of the bacteria. These findings thus describe a novel immunoevasion pathway engaged by Mtb. The central hypothesis of this application is that genetic screens will be able to identify the genes in Mtb important for inhibition of the IFNAR-mediated cell signaling. In Aim 1.1 we propose to execute a “gain-of-function” genetic screen using M. smegmatis clones transfected with a Mtb-DNA cosmid library. This approach has been successfully executed for two other screens by the PI’s lab. Next, in Aim 1.2 we describe how the relevant Mtb gene out of the approximately 40 genes in one of the isolated cosmids will be identified. Overall, we anticipate that the successful completion of the current project will generate at least one Mtb deletion mutant deficient in inhibition of IFNAR-mediated cell signaling and potentially the knowledge of several other Mtb regions that contain additional genes. These findings will be the basis for a competitive R01 application which will propose to: 1) complete the identification of all of the Mtb genes involved in inhibition of IFN-β-mediated cell signaling, 2) to use the mutants for detailed analysis of the molecular mechanisms of the inhibition and 3) to characterize the importance of inhibiting IFNAR-mediated cell signaling for in vivo virulence of Mtb.

抽象的据估计，世界上约三分之一的人口潜伏感染结核分枝杆菌 (Mtb) 每年新增 1000 万结核病病例，导致约 100 万人死亡，这进一步凸显了这一点。这种病原体对全球公共卫生的影响已经发展出多种逃避宿主的途径。在大多数情况下，这些宿主病原体相互作用的分子机制只是。当前提案的前提得到了一系列新颖和新颖的支持。未发表的结果表明 Mtb 抑制 IFN-a/b 受体 (IFNAR) 介导的细胞信号传导，结果改善了细菌的细胞内生长，因此这些发现描述了一种新的免疫逃避。 Mtb 参与的途径该应用的中心假设是基因筛选将能够识别在目标 1.1 中，我们建议执行 Mtb 中对抑制 IFNAR 介导的细胞信号转导很重要的基因。使用 Mtb-DNA 粘粒文库转染的耻垢分枝杆菌克隆进行“功能获得”遗传筛选。 PI 实验室已在另外两个屏幕上成功执行了该方法，接下来，我们在目标 1.2 中进行了描述。如何从分离的粘粒之一的大约 40 个基因中鉴定出相关的 Mtb 基因。总体而言，我们预计当前项目的成功完成将产生至少一个 Mtb 删除突变体缺乏对 IFNAR 介导的细胞信号传导的抑制，并且可能缺乏对其他几种 Mtb 的了解这些发现将成为竞争性 R01 应用的基础。将提出：1）完成所有参与抑制 IFN-β 介导细胞的 Mtb 基因的鉴定信号传导，2) 使用突变体详细分析抑制的分子机制，3) 描述了抑制 IFNAR 介导的细胞信号传导对于 Mtb 体内毒力的重要性。