Genetic Dissection of Mycobacterial Pathogenesis During Eicosanoid-Mediated Immunity

类花生酸介导的免疫过程中分枝杆菌发病机制的遗传解析

• 批准号: 10314405
• 负责人: Erika Joy Hughes
• 金额: $ 3.8万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10314405
• 关键词: Ablation; Affect; Anti-Inflammatory Agents; Attenuated; Bacterial Genes; Binding; Biology; Cause of Death; Cell physiology; Cells; Complex; Data; Data Set; Disease; Dissection; Eicosanoids; Equilibrium; Fluorescence; Gene Expression Profile; Genes; Genetic; Genetic study; Genome; Genus Mycobacterium; Granuloma; Granulomatous; Growth; Histology; Human; Image; Immune; Immunity; Infection; Infection Control; Infectious Agent; Inflammatory; Libraries; Lipids; Maps; Mediating; Methods; Modeling; Monitor; Mus; Mutant Strains Mice; Mutation; Mycobacterium Infections; Mycobacterium marinum; Mycobacterium tuberculosis; Myelogenous; Outcome; Pathogenesis; Phenotype; Population; Prevention; Production; Proteins; Resistance; Role; Shapes; Side; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Structure; System; Testing; Therapeutic; Tuberculosis; Virulence; Zebrafish; apolipoprotein D; base; cell type; differential expression; experimental study; genetic approach; in vivo; infection burden; leukotriene A4 hydrolase; macrophage; mature animal; mouse model; mutant; mycobacterial; novel therapeutics; pathogen; pressure; programs; response; single-cell RNA sequencing; small molecule; transcriptome; transposon sequencing

ABSTRACT Mycobacterium tuberculosis (Mtb) one of the most successful pathogen in the world due to its long-standing, ancient partnership with humans. Despite decades of effort put towards prevention and treatment, Mtb was the number one cause of death by any single infectious agent in 2019. An important regulator of mycobacterial infections are eicosanoids: host-produced, lipid signaling molecules that can drive both inflammatory and anti- inflammatory signaling cascades. Using a zebrafish-Mycobacterium marinum infection model that recapitulates important aspects of mycobacterial pathogenesis, the balance of specific pro- and anti-inflammatory eicosanoids was found to be critical to control of infection. Disruption of eicosanoid synthesis, through a mutation in leukotriene A4 hydrolase-deficient (lta4h-/-), results in altered granuloma structure. The granuloma is the central feature of Mtb infection and is an aggregation of immune cells coordinating to restrict bacterial growth. This is the principal site of host-pathogen interactions. Although the granuloma is a key part of tuberculosis pathogenesis, host-pathogen signaling programs that govern granuloma biology remain elusive. Our lack of understanding of granuloma formation and signaling is due to the difficulty in studying the granuloma because the complex cell organization can only be formed and observed in vivo. The zebrafish-Mycobacterium marinum model, with conserved virulence loci and host-pathogen genetic programs, is used to study the genetics of mycobacterial infection and granuloma structure. This proposal will use genetic approaches in both the zebrafish- M.marinum granuloma model and the mouse-Mtb model of infection to test the hypothesis that there are key differences in eicosanoid-mediated immunity and bacterial response that are integral to overall granuloma biology. To this end, the first single-cell map of mycobacterial granulomas, in an experimentally tractable system was assembled from wildtype and lta4h-/- zebrafish granuloma cells. The findings from this dataset informed the following approaches to interrogate the effects of host eicosanoid signaling on granuloma structure and mycobacterial response. Aim 1 will define the functional role of a previously undefined population of granuloma cells that are dependent on lta4h expression. Aim 2 will use Mtb TnSeq approaches to the examine the genetic requirements of Mtb in mice that lack key components of eicosanoid signaling. Together, these aims will enhance our understanding of granuloma biology and mycobacterial response and can help guide therapeutic approaches to treating tuberculosis.

摘要 结核分枝杆菌（Mycobacterium tuberculosis，Mtb）是世界上最成功的病原体之一， 与人类的古老伙伴关系。尽管几十年来在预防和治疗方面做出了努力，但结核病仍是 2019年任何单一传染性病原体导致的头号死亡原因。一种重要的分枝杆菌调节因子 感染是类花生酸：宿主产生的脂质信号分子，可以驱动炎症和抗 炎症信号级联。使用斑马鱼-海洋分枝杆菌感染模型， 分枝杆菌发病机制的重要方面，特异性促炎和抗炎类花生酸的平衡 对控制感染至关重要。类花生酸合成的破坏，通过突变， 白三烯A4水解酶缺乏（lta 4 h-/-），导致肉芽肿结构改变。肉芽肿位于 Mtb感染的特征，并且是协调以限制细菌生长的免疫细胞的聚集。这是 宿主-病原体相互作用的主要场所。虽然肉芽肿是结核病的关键部位 然而，在发病机制中，控制肉芽肿生物学的宿主-病原体信号传导程序仍然难以捉摸。我们缺乏 对肉芽肿形成和信号传导的理解是由于研究肉芽肿的困难， 复杂的细胞组织只能在体内形成和观察。斑马鱼-海洋分枝杆菌 模型，保守的毒力基因座和宿主-病原体遗传程序，用于研究的遗传学， 分枝杆菌感染和肉芽肿结构。这项提议将在斑马鱼和斑马鱼中使用遗传方法- M.marinum肉芽肿模型和小鼠-Mtb感染模型来检验存在关键的 类花生酸介导的免疫和细菌反应的差异， 肉芽肿生物学为此，在一个实验中， 从野生型和LTA 4 H-/-斑马鱼肉芽肿细胞组装易处理的系统。这一发现 数据集提供了以下方法来询问宿主类花生酸信号传导对肉芽肿的影响 结构和分枝杆菌反应。目标1将界定以前未界定的人口的职能作用 依赖于LTA 4 H表达的肉芽肿细胞。目标2将使用Mtb TnSeq方法来检查 Mtb在缺乏类花生酸信号传导关键组分的小鼠中的遗传要求。总之，这些目标 将提高我们对肉芽肿生物学和分枝杆菌反应的理解， 肺结核的治疗方法。