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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要结核分枝杆菌是结核病的病原体，仍然是发病和死亡的重要原因在世界范围内， oma 内持续存在颗粒缺氧状况是结核病的标志，导致潜伏感染。并限制当前疗法的应用。了解结核分枝杆菌如何支持其代谢。在缺氧条件下并适应这些挑战是消除潜在结核病“解除武装”的关键。通过消除其耐受肉芽肿中应激条件的能力来治疗结核病将提供一种可行的方法针对潜伏性结核病的临床干预策略。我们将研究辅酶 F420 在结核分枝杆菌 F420 的生理学和发病机制中的作用。越来越多的证据表明，脱氮黄素在多种氢化物转移反应中充当载体。结核分枝杆菌广泛使用这种辅助因子，其氧化还原电位低，被认为使其在缺氧中发挥关键作用分枝杆菌中似乎存在 F420 依赖性机制，可用于预防。氧化和亚硝化应激，但 F420 对结核分枝杆菌代谢和持久性的贡献目前尚不清楚该项目的最终目标是研究 F420 如何促进结核分枝杆菌的存活。在肉芽肿内的缺氧条件下，阐明其在持久性中的作用我们设计了一种综合方法。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为了确定 F420 如何帮助结核分枝杆菌在发病过程中持续存在，我们将构建一组条件。针对 F420 生物合成和代谢的敲低菌株，并进行体外研究以确定 M. 结核病在缺氧条件下和缺氧诱导休眠后的重新生长过程中利用F420。然后在出现坏死和缺氧结核病灶的小鼠模型中进行感染研究以进行调查 F420 如何促进结核分枝杆菌在肉芽肿内存活并了解 F420 与结核分枝杆菌之间的联系最后，我们将揭示 F420 发色团生物合成的结构见解，提供 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。