Coenzyme F420, helping mycobacteria find a niche in humans

辅酶 F420，帮助分枝杆菌在人类中找到一席之地

• 批准号: 10666639
• 负责人: Michael Berney
• 金额: $ 19.8万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10666639
• 关键词: Adopted; Aerobic Bacteria; Anabolism; Anaerobic Bacteria; Animals; Attention; Autophagocytosis; Biochemical Pathway; Biochemistry; Biology; Biosynthetic Proteins; C3HeB/FeJ Mouse; Clinical; Coenzymes; Disease; Drug Tolerance; Ensure; Enzymes; Evolution; Flavins; Flavoproteins; Foundations; Genetic; Genome; Genus Mycobacterium; Goals; Granuloma; Growth; Host Defense; Human; Hypoxia; Immune Evasion; Immune system; In Vitro; Infection; International; Intervention; Lesion; Life Style; Link; M. tuberculosis genome; Macrophage; Metabolic; Metabolism; Modeling; Morbidity - disease rate; Mycobacterium leprae; Mycobacterium smegmatis; Mycobacterium tuberculosis; Necrosis; Niacinamide; Nitroimidazoles; Organ; Oxidation-Reduction; Oxidative Stress; Oxygen; Pathogenesis; Pathology; Pathway interactions; Pharmaceutical Preparations; Physiology; Play; Point Mutation; Predisposition; Prodrugs; Production; Protein Biochemistry; Proteins; Reaction; Research; Research Personnel; Role; S-Adenosylhomocysteine; S-Adenosylmethionine; Stress; Structure; System; Time; Tuberculosis; biochemical tools; chromophore; cofactor; environmental stressor; experience; improved; in vitro Model; insight; knock-down; latent infection; metabolomics; microorganism; mortality; mouse model; mutant; mycobacterial; nitrosative stress; pathogen; programs; structural biology; transcriptomics; tuberculosis treatment

Project summary Mycobacterium tuberculosis, the causative agent of TB, remains an important cause of morbidity and mortality worldwide. Persistence in hypoxic conditions within granuloma is the hallmark of TB, leading to latent infection and limiting the application of current therapies. Understanding how M. tuberculosis supports its metabolism under hypoxic conditions, and adapts to these challenges, is key to eliminating latent TB. “Disarming” M. tuberculosis by removing its ability to endure the stress-inducing conditions in granuloma will provide a feasible strategy for clinical interventions against latent TB. We will investigate the role of the coenzyme F420 in the physiology and pathogenesis of M. tuberculosis. F420 is a deazaflavin that acts as a carrier in a wide range of hydride transfer reactions. Growing evidence points to the wide use of this cofactor by M. tuberculosis, with its low redox potential thought to give it key roles in hypoxic persistence. There appears to be F420-dependent mechanisms in mycobacteria that are used to protect against oxidative and nitrosative stress, but the contribution of F420 to M. tuberculosis metabolism and persistence remains unclear. The ultimate goal of the project is to investigate how F420 facilitates M. tuberculosis’ survival under hypoxic conditions within granuloma, elucidating its role in persistence. We have devised an integrated experimental approach using genetics, metabolomics, animal infection models, and protein biochemistry tools to determine how F420 helps M. tuberculosis persist during pathogenesis. We will construct a set of conditional knockdown strains targeting F420 biosynthesis and metabolism, and perform in vitro studies to determine how M. tuberculosis utilizes F420 under hypoxic conditions and during re-growth after hypoxia-induced dormancy. We will then perform infection studies in a mouse model that develops necrotic and hypoxic TB lesions to investigate how F420 facilitates M. tuberculosis survival inside granuloma and to understand the link between F420 and persistence. Finally, we will reveal structural insights into the biosynthesis of F420 chromophore, providing the foundations for mechanistic and inhibition studies of this essential reaction in F420 biosynthesis.

项目摘要 结核分枝杆菌是结核病的病原体，仍然是发病率和死亡率的重要原因 国际吧肉芽肿内持续缺氧是结核病的标志，导致潜伏感染 并限制了当前疗法的应用。了解M。结核病支持它的新陈代谢 在低氧条件下，并适应这些挑战，是消除潜伏性结核病的关键。"解除武装" M 结核病通过消除其能力，以承受应力诱导条件下的肉芽肿将提供一个可行的 针对潜伏性结核病临床干预战略。 我们将研究辅酶F420在M.结核F420是 一种在广泛的氢化物转移反应中作为载体的脱氮黄素。越来越多的证据表明， M广泛使用该辅因子。结核病，其低氧化还原电位被认为在缺氧中发挥关键作用， 坚持不懈在分枝杆菌中似乎存在F420依赖性机制，用于防止 氧化和亚硝化胁迫，但F420对M.结核代谢和持久性 仍不清楚本项目的最终目的是研究F420是如何促进M。结核病生存率 在缺氧条件下肉芽肿，阐明其作用的持久性。我们设计了一个综合的 使用遗传学、代谢组学、动物感染模型和蛋白质生物化学工具的实验方法 以确定F420如何帮助M。结核病在发病过程中持续存在。我们将构建一组条件 敲减菌株靶向F420的生物合成和代谢，并进行体外研究，以确定如何M。 结核杆菌在缺氧条件下和在缺氧诱导的休眠后的再生长期间利用F420。我们将 然后在小鼠模型中进行感染研究，该模型产生坏死和缺氧的结核病病变， F420如何促进M.肉芽肿内结核病的生存，并了解F420和 坚持不懈最后，我们将揭示F420发色团生物合成的结构见解， 为F420生物合成中这一重要反应的机理和抑制研究奠定了基础。