Regulators of IFN-gamma responses during Mycobacterium tuberculosis infection

结核分枝杆菌感染期间 IFN-γ 反应的调节因子

• 批准号: 10659240
• 负责人: Andrew Olive
• 金额: $ 52.4万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-06 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659240
• 关键词: Address; Antigen Presentation; Antigen-Presenting Cells; Automobile Driving; Bacterial Model; CD4 Positive T Lymphocytes; CRISPR screen; Cause of Death; Cell Communication; Cell physiology; Cells; Cessation of life; Chemicals; Chronic; Communicable Diseases; Complex; Coupled; Data; Development; Disease; Disease Progression; Disputes; Epidemic; Equilibrium; Genes; Genetic Screening; Glycerol; Glycogen Synthase Kinases; Goals; Growth; Immune; Immune Evasion; Immune response; Immunity; Infection; Inflammation; Inflammatory; Inflammatory Response; Innate Immune Response; Interferon Type II; Kinetics; Knowledge; Lead; Lung; Lung diseases; Macrophage; Macrophage Activation; Mediating; Mediator; Metabolic; Mus; Mycobacterium tuberculosis; Myelogenous; Outcome; Pathogenesis; Pathologic; Pathway interactions; Phosphotransferases; Play; Production; Public Health; Regulation; Role; Shapes; Signal Transduction; Structure of parenchyma of lung; Surface; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Time; Tissues; Transgenic Organisms; Tuberculosis; Vaccines; Virulence; adaptive immune response; cytokine; effective therapy; effector T cell; genetic approach; genome-wide; in vivo; in vivo Model; interdisciplinary approach; novel therapeutics; paralogous gene; pathogen; prevent; protective pathway; response; targeted treatment; transcriptome

Summary Tuberculosis (TB) is a leading cause of death by infectious disease worldwide. Current therapy requires at least six months of treatment and there is no effective vaccine that prevents pulmonary disease. There is a critical need to develop new therapies that will shorten treatment time and activate protective responses to prevent TB progression. Unfortunately, we continue to lack a fundamental understanding of the host immune pathways that protect against TB. While the cytokine interferon γ (IFNγ) is absolutely required, the precise mechanisms that drive IFNγ-mediated protection remain unclear. This is because IFNγ is a pleotropic mediator that controls both bacterial growth and the inflammatory response in the lungs, while bridging the innate and adaptive immune responses. To dissect the complex regulation of IFNγ responses, we conducted a genome-wide CRISPR Cas9 screens in macrophages to define the networks required for the IFNγ-mediated surface expression of MHCII. This screen uncovered an important role for the glycerol 3 kinase beta (GSK3b) in controlling IFNγ-dependent MHCII expression. Using chemical and genetic approaches coupled with transcriptome analysis we found that GSK3b and the paralog GSK3a contribute to the expression of a subset of IFNγ-inducible genes. Further mechanistic studies found that the loss of GSK3a/b blocks the ability of macrophages to serve as antigen presenting cells to CD4+ T cells ex vivo and results in dysregulated cytokine production by macrophages during M. tuberculosis infection. Thus, GSK3a/b balances the cellular response to IFNγ to protect against infection without driving inflammatory tissue damage. These data lead to the hypothesis that GSK3a/b is a critical mediator of the IFNγ response during TB in vivo. This proposal will use genetic approaches combined with in vivo models to dissect the role of GSK3a/b-mediated control of IFNγ responses and TB. Aim 1 will understand the mechanisms that regulate GSK3a/b control of IFNγ responses and determine how M. tuberculosis modulates GSK3a/b activity. In Aim 2, we will use unique models of bacterial control and inflammation to examine how the loss of myeloid-specific GSK3a and/or GSK3b disrupts the IFNγ-mediated control of M. tuberculosis growth and tissue damage. Finally, in Aim 3 we will use M. tuberculosis specific TCR-transgenic T cell mice to test the hypothesis that the loss of myeloid GSK3a and/or GSK3b results in reduced T cell responses and ineffective protection against TB. Together these aims will dissect the role of GSK3a/b in controlling IFNγ-responses and protecting against TB. These mechanisms can then be leveraged to develop new therapies that may shorten treatment times and prevent TB disease progression.

概括 结核病 (TB) 是全球传染病导致死亡的主要原因，目前的治疗至少需要六种治疗方法。 几个月的治疗，并且没有有效的疫苗可以预防肺部疾病。 开发新疗法，缩短治疗时间并激活保护反应以防止结核病进展。 不幸的是，我们仍然缺乏对宿主免疫途径的基本了解，以防止 虽然细胞因子干扰素 γ (IFNγ) 是绝对必需的，但驱动 IFNγ 介导的精确机制 这是因为 IFNγ 是一种多效性介质，既控制细菌生长又控制细菌生长。 肺部炎症反应，同时桥接先天性和适应性免疫反应。 由于 IFNγ 反应的复杂调控，我们在巨噬细胞中进行了全基因组 CRISPR Cas9 筛选，以 定义了 IFNγ 介导的 MHCII 表面表达所需的网络。该筛选揭示了一个重要的信息。 使用化学和方法研究甘油 3 激酶 β (GSK3b) 在控制 IFNγ 依赖性 MHCII 表达中的作用。 遗传方法与转录组分析相结合，我们发现 GSK3b 和旁系同源 GSK3a 有助于 进一步的机制研究发现，IFNγ诱导基因的一个子集的表达缺失。 GSK3a/b 阻断巨噬细胞作为体外 CD4+ T 细胞抗原呈递细胞的能力和结果 因此，GSK3a/b 可以平衡结核分枝杆菌感染期间巨噬细胞产生的细胞因子的失调。 这些数据导致细胞对 IFNγ 的反应，以防止感染而不引起炎症组织损伤。 本提案将使用 GSK3a/b 是体内 TB 期间 IFNγ 反应的关键介质的假设。 遗传方法与体内模型相结合，剖析 GSK3a/b 介导的 IFNγ 反应控制的作用 目标 1 将了解调节 GSK3a/b 控制 IFNγ 反应的机制并确定如何调节。 结核分枝杆菌调节 GSK3a/b 活性 在目标 2 中，我们将使用独特的细菌控制和炎症模型。 研究骨髓特异性 GSK3a 和/或 GSK3b 的缺失如何破坏 IFNγ 介导的分枝杆菌控制。 最后，在目标 3 中，我们将使用结核分枝杆菌特异性 TCR 转基因 T 细胞。 小鼠测试髓系 GSK3a 和/或 GSK3b 缺失导致 T 细胞反应降低的假设 这些目标将共同剖析 GSK3a/b 在控制 IFNγ 反应中的作用。 然后可以利用这些机制来开发可能缩短时间的新疗法。 治疗时间并防止结核病进展。