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.

摘要 结核病(TB)是世界范围内主要的传染病死亡原因。目前的治疗方法至少需要六个 几个月的治疗，而且没有有效的疫苗来预防肺部疾病。迫切需要 开发新的治疗方法，缩短治疗时间并激活保护性反应，以防止结核病的进展。 不幸的是，我们仍然缺乏对宿主免疫途径的基本了解，这些免疫途径可以防止 结核病。虽然细胞因子干扰素γ(干扰素γ)是绝对必需的，但驱动干扰素γ介导的确切机制 保护措施仍不明朗。这是因为干扰素γ是一种多发性介质，它既控制细菌的生长，又控制 肺部的炎症反应，同时连接先天和获得性免疫反应。要仔细分析 干扰素γ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 确定干扰素γ介导的MHCII表面表达所需的网络。这个屏幕揭示了一个重要的 甘油3激酶β(GSK3b)在控制干扰素γ依赖的MHCII表达中的作用使用化学药品和 遗传方法结合转录组分析，我们发现GSK3b和Paralog GSK3a对 干扰素γ诱导基因亚集的表达。进一步的机制研究发现， GSK3a/b体外阻断巨噬细胞作为CD4+T细胞抗原提呈细胞的能力和结果 在结核分枝杆菌感染过程中巨噬细胞产生细胞因子的失调。因此，GSK3a/b平衡了 细胞对干扰素γ的反应，以防止感染，而不会导致炎症组织损伤。这些数据导致 假设GSK3a/b是体内结核病过程中干扰素γ反应的关键介质。这项提案将使用 遗传学方法结合体内模型剖析GSK3a/b介导的干扰素γ应答的调控作用 和肺结核。目标1将了解调节GSK3a/b控制干扰素γ反应的机制，并确定如何 结核分枝杆菌调节GSK3a/b的活性。在目标2中，我们将使用独特的细菌控制和炎症模型 研究髓系特异性GSK3a和/或GSK3b的缺失如何干扰干扰素γ介导的M。 肺结核的生长和组织损伤。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 小鼠验证髓系GSK3a和/或GSK3b丢失导致T细胞反应减少和 对结核病的保护不力。这些目标将共同剖析GSK3a/b在控制干扰素γ反应中的作用 以及预防结核病。然后可以利用这些机制来开发新的疗法，这些疗法可能会缩短 治疗次数和防止结核病的进展。