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.

项目摘要结核分枝杆菌是结核病的病因，仍然是发病率和死亡率的重要原因全世界。肉芽肿内缺氧条件的持久性是结核病的标志，导致潜在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新陈代谢在低氧条件下，适应这些挑战是消除潜在结核病的关键。 “解除武装” M。结核病通过消除其忍受应力诱导条件的肉芽肿的能力将提供可行的针对潜在结核的临床干预策略。我们将研究辅酶F420在结核分枝杆菌的生理和发病机理中的作用。 F420是 Deazaflavin在较广泛的氢化物转移反应中充当载体。越来越多的证据指向结核分枝杆菌对这种辅助因子的广泛使用，其低氧化还原潜力被认为在低氧中发挥了关键作用持久性。分枝杆菌中似乎有F420依赖性机制用于预防氧化和亚硝化应激，但F420对结核分枝杆菌的代谢和持久性的贡献仍然不清楚。该项目的最终目的是研究F420如何促进结核分枝杆菌的生存在颗粒瘤内的低氧条件下，阐明了其在持久性中的作用。我们设计了一个集成的实验方法使用遗传学，代谢组学，动物感染模型和蛋白质生物化学工具为了确定F420如何帮助结核分枝杆菌在发病机理期间持续存在。我们将构建一组有条件的针对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在缺氧引起的休眠状态后重生期间利用F420。我们将然后在小鼠模型中进行感染研究，该模型发展了坏死和低氧TB病变 F420如何促进肉芽肿内的结核分枝杆菌生存，并了解F420和F420之间的联系持久性。最后，我们将揭示对F420发色团生物合成的结构见解，从而提供 F420生物合成中这种基本反应的机械和抑制研究基础。