Hypoxia, tuberculosis, and T cell dysfunction

缺氧、结核和 T 细胞功能障碍

• 批准号: 10735553
• 负责人: SAMUEL M BEHAR
• 金额: $ 72.97万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735553
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Alcoholism; Animal Model; Antibodies; Bacillus; Bacteria; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cell physiology; Cell surface; Cellular Immunity; Cellular Metabolic Process; Cessation of life; Chronic; Clinical; Containment; Coupled; Data; Development; Diabetes Mellitus; Disease; Equilibrium; Failure; Flow Cytometry; Functional disorder; Goals; Granuloma; Growth; Homeostasis; Human; Hypoxia; Immune; Immunity; Immunooncology; Impairment; In Vitro; Infection; Intervention; Knowledge; Lead; Lesion; Lung; Malignant Neoplasms; Malnutrition; Maps; Mediating; Metabolic; Metabolic stress; Metabolism; Mitochondria; Modeling; Mouse Strains; Mus; Mycobacterium tuberculosis; Outcome; Pathogenesis; Patients; Persons; Pharmaceutical Preparations; Phenotype; Prediction of Response to Therapy; Predisposition; Proliferating; Pulmonary Tuberculosis; Receptor Signaling; Recrudescences; Reporter; Resistance; Resolution; Risk; Risk Factors; Role; Smoking; T cell receptor repertoire sequencing; T cell response; T-Cell Activation; T-Cell Development; T-Lymphocyte; Testing; Therapeutic; Tuberculosis; Tumor Immunity; Tumor-Infiltrating Lymphocytes; Work; cancer therapy; exhaust; exhaustion; experimental study; global health; improved; in vivo; insight; mouse model; neovascularization; preservation; prevent; programmed cell death protein 1; prophylactic; receptor; stressor; tumor; tumor microenvironment; Acquired Immunodeficiency Syndrome; Affect; Alcoholism; Animal Model; Antibodies; Bacillus; Bacteria; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cell physiology; Cell surface; Cellular Immunity; Cellular Metabolic Process; Cessation of life; Chronic; Clinical; Containment; Coupled; Data; Development; Diabetes Mellitus; Disease; Equilibrium; Failure; Flow Cytometry; Functional disorder; Goals; Granuloma; Growth; Homeostasis; Human; Hypoxia; Immune; Immunity; Immunooncology; Impairment; In Vitro; Infection; Intervention; Knowledge; Lead; Lesion; Lung; Malignant Neoplasms; Malnutrition; Maps; Mediating; Metabolic; Metabolic stress; Metabolism; Mitochondria; Modeling; Mouse Strains; Mus; Mycobacterium tuberculosis; Outcome; Pathogenesis; Patients; Persons; Pharmaceutical Preparations; Phenotype; Prediction of Response to Therapy; Predisposition; Proliferating; Pulmonary Tuberculosis; Receptor Signaling; Recrudescences; Reporter; Resistance; Resolution; Risk; Risk Factors; Role; Smoking; T cell receptor repertoire sequencing; T cell response; T-Cell Activation; T-Cell Development; T-Lymphocyte; Testing; Therapeutic; Tuberculosis; Tumor Immunity; Tumor-Infiltrating Lymphocytes; Work; cancer therapy; exhaust; exhaustion; experimental study; global health; improved; in vivo; insight; mouse model; neovascularization; preservation; prevent; programmed cell death protein 1; prophylactic; receptor; stressor; tumor; tumor microenvironment

Abstract. After Mycobacterium tuberculosis (Mtb) infection, 5-10% of people develop clinically evident tuberculosis (TB), most within two years. This leads to 10 million new cases of TB and 1.5 million deaths each year. Why immunity fails and permits recrudescence in people that initially control Mtb is unknown. Risk factors include diabetes, malnutrition, alcoholism, cancer, and smoking, all which cause metabolic stress. Our long-term goal is to understand the drivers of immune failure and identify protective mechanisms of immunity. A major knowledge gap is how various metabolic insults affect cellular immunity in the infected lung. Our over-arching hypothesis is that that during TB, metabolic stressors such as granuloma hypoxia contribute to T cell dysfunction, degrade immunity, and impair Mtb containment. We and others find that T cells from patients with pulmonary TB and chronically Mtb-infected mice are dysfunctional. Dysfunctional CD8 T cells (e.g., exhausted CD8 T cells) have been intensively studied because of their role in tumor immunity. In contrast, far less is known about CD4 T cell dysfunction. We will investigate both CD4 and CD8 T cells and focus on CD4 T cells as they are crucial for immunity to Mtb. We will use the murine TB model to investigate how metabolic stress affects T cell function and contributes to TB pathogenesis. An important component of our strategy is to compare T cells from susceptible mice that develop hypoxic granulomas with T cells from resistant mouse strains. The first aim is to “Determine the relationship between metabolic perturbation and T cell dysfunction.” A high-resolution map of T cell responses to Mtb in susceptible and resistant mice will be assembled after performing scRNASeq, TCRseq, conventional flow cytometry and MetFlow (to assess cell metabolism). We will determine whether dysfunctional T cells differ in their control of Mtb in vitro and in vivo. Secondly, we will “Determine how hypoxia affects T cell immunity against Mtb.” Using hypoxia fate reporter mice, we will study how hypoxia affects T cell function in vivo. These studies will be coupled with mechanistic studies using hypoxic culture conditions in vitro. We will establish how hypoxia, metabolic stress, and T cell function are related, and whether hypoxia is detrimental to protective T cell responses during TB. Finally, Aim 3 will “Assess how metabolic interventions alter T cell function and TB outcome.” We predict that drugs that correct underlying metabolic perturbations can improve T cell function and enhance control of Mtb infection. Using the hypoxic mouse models, proof-of-principle experiments will be done to determine how drugs that affect neovascularization, target metabolism, or protect mitochondria, affect Mtb containment in vivo. Our studies will determine how hypoxia and metabolic stress affect immunity to Mtb and provide insight into why CD4 immunity fails. As T cells are essential in containing Mtb infection, we hypothesize that interventions to limit T cell dysfunction or improve their function will augment Mtb containment. Metabolic reprogramming of T cells in the tumor microenvironment is on the horizon. Our hypothesis embraces the idea that metabolic therapeutics could prevent or reverse T cell dysfunction and improve TB outcome.

抽象的。结核分枝杆菌感染（MTB）感染后，有5-10％的人出现临床证据 结核病（TB），大部分在两年内。这会导致1000万例新的结核病和150万例 年。为什么免疫失败并允许最初控制MTB的人重新发明。风险因素 包括糖尿病，营养不良，酒精中毒，癌症和吸烟，所有这些都会导致代谢压力。我们的长期 目标是了解免疫失败的驱动因素，并确定免疫史的受保护机制。专业 知识差距是各种代谢损伤如何影响感染肺的细胞免疫。我们的整理 假设是，在结核病期间，代谢胁迫（例如肉芽肿缺氧）导致T细胞功能障碍， 降解免疫力并损害MTB遏制。我们和其他人发现来自肺结核患者的T细胞 慢性MTB感染的小鼠功能失调。功能失调的CD8 T细胞（例如，耗尽的CD8 T细胞） 由于它们在肿瘤免疫学中的作用，因此被深入研究。相反，关于CD4的知之甚少 T细胞功能障碍。我们将研究CD4和CD8 T细胞，并专注于CD4 T细胞，因为它们至关重要 为了免疫MTB。我们将使用鼠结核模型来研究代谢应力如何影响T细胞功能 并有助于结核病发病机理。我们策略的重要组成部分是比较来自 易感小鼠，与抗性小鼠菌株的T细胞发育低氧肉芽肿。第一个目的是 “确定代谢扰动与T细胞功能障碍之间的关系。” T的高分辨率图 在执行scrnaseq，tcrseq， 常规的流式细胞仪和METFlow（评估细胞代谢）。我们将确定功能失调 T细胞在体外和体内对MTB的控制方面有所不同。其次，我们将“确定缺氧如何影响T细胞 对MTB的免疫力。“使用缺氧脂肪报告小鼠，我们将研究缺氧如何影响体内T细胞功能。 这些研究将在体外使用低氧培养条件与机械研究结合使用。我们将建立 缺氧，代谢应激和T细胞功能如何相关，以及缺氧是否有害保护 TB期间的T细胞反应。最后，AIM 3将“评估代谢干预如何改变T细胞功能和TB 结果。我们预测，纠正潜在代谢扰动的药物可以改善T细胞功能和 增强对MTB感染的控制。使用低氧鼠标模型，将进行原理证明实验 确定影响新血管形成，靶向代谢或保护线粒体的药物如何影响MTB 体内遏制。我们的研究将确定缺氧和代谢应激如何影响对MTB和MTB的免疫学 提供有关为什么CD4免疫疾病失败的见解。由于T细胞对于包含MTB感染至关重要，我们假设 限制T细胞功能障碍或改善其功能的干预措施将增加MTB的控制。代谢 肿瘤微环境中T细胞的重编程即将到来。我们的假设包含了这个想法 这种代谢疗法可以预防或逆转T细胞功能障碍并改善结核病结果。