Metabolomic Analysis of Redox control in Mycobacterium tuberculosis persisters

结核分枝杆菌持续存在氧化还原控制的代谢组学分析

• 批准号: 7980241
• 负责人: WILLIAM Robert JACOBS
• 金额: $ 24.9万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7980241
• 关键词: Aerobic; Aerobiosis; Affect; Anaerobic Bacteria; Anaerobiosis; Antitubercular Agents; Bacillus (bacterium); Bacteria; Biochemical; Biological Models; Butyrates; Carbon; Carbon Dioxide; Cause of Death; Cells; Citric Acid Cycle; Complement; Cytochrome c Reductase; Decarboxylation; Development; Drug resistance; Drug resistance in tuberculosis; Effectiveness; Enzymes; Ethionamide; Exposure to; Extreme drug resistant tuberculosis; Fatty Acids; Genes; Genetic; Glucose; Glycerol; Goals; Growth; HIV; Hour; Hypoxia; Knowledge; Lead; Life; Link; Mass Spectrum Analysis; Mediating; Metabolic; Metabolic Control; Metabolism; Microbial Biofilms; Modeling; Multi-Drug Resistance; Mutation; Mycobacterium tuberculosis; Opportunistic Infections; Organism; Oxidation-Reduction; Oxidoreductase; Pathway interactions; Pharmaceutical Preparations; Phenotype; Physiological; Population Heterogeneity; Protein Biosynthesis; Resistance; Rifampin; Role; Route; Starvation; Time; Treatment Efficacy; Tuberculosis; Water; bactericide; base; chemotherapy; improved; isoniazid; killings; metabolomics; mutant; novel; novel strategies; pandemic disease; prevent; public health relevance; stable isotope; tuberculosis drugs

DESCRIPTION (provided by applicant): New chemotherapeutics are urgently required to control the tuberculosis pandemic, which is fueled by the emergence of multi-drug- and extensively-drug-resistant Mycobacterium tuberculosis strains and the bacterium's catastrophic alliance with HIV. In addition to genetic resistance to drugs, M. tuberculosis cells can become persistors, cells that are phenotypically resistant to the killing by drugs. This ability to enter into a persistent state necessitates long periods of chemotherapy and represents the greatest challenge in improve TB chemotherapies. Unfortunately, little is known about the metabolic state of persistors. To elucidate the physiological state of persistent M. tuberculosis cells, we have used mass spectroscopy to generate metabolic profiles of M. tuberculosis when grown in aerobic and hypoxic growth conditions (a persistence model) and found several key control points modulating NADH/NAD (redox) metabolism. To further characterize the metabolic profiles, we have investigated the Tricarboxylic Acid Cycle (TCA) of M. tuberculosis. Using a combination of genetic and biochemical approaches, we have discovered that M. tuberculosis possesses a set of genes encoding ketoglutarate oxidoreductase, a key enzyme of the TCA that is generally observed in anaerobic bacteria. In addition, we have discovered that mutations in the gene encoding NADH dehydrogenase II (ndh) confer resistance to Isoniazid and Ethionamide. By combining metabolite profiling, and stable isotope flux profiling, with mutational studies we intend to characterize persistent states of M. tuberculosis. This knowledge should lead to novel strategies to effectively kill persistent M. tuberculosis cells. PUBLIC HEALTH RELEVANCE: New chemotherapeutics are urgently required to control the tuberculosis pandemic, which is fueled by the emergence of multi-drug- and extensively-drug-resistant Mycobacterium tuberculosis strains and the bacterium's catastrophic alliance with HIV. In addition to genetic resistance to drugs, M. tuberculosis cells can become persistors, cells that are phenotypically resistant to the killing by drugs. This proposal seeks to develop new understandings of persisting M. tuberculosis cells providing the knowledge for novel and shortened chemotherapies.

描述（由申请人提供）：迫切需要新的化疗药物来控制结核病大流行，这是由多重耐药和广泛耐药结核分枝杆菌菌株的出现以及该细菌与HIV的灾难性联盟所推动的。除了对药物的遗传抗性，M.结核病细胞可以成为持续存在的细胞，这些细胞在表型上对药物的杀伤具有抗性。这种进入持久状态的能力需要长期化疗，并且是改善结核病化疗的最大挑战。不幸的是，很少有人知道的代谢状态的坚持。阐明宿存M.结核杆菌细胞中，我们已经使用质谱产生M的代谢谱。在有氧和低氧生长条件下生长时，结核杆菌的生长（持久性模型），并发现了几个调节NADH/NAD（氧化还原）代谢的关键控制点。为了进一步表征代谢谱，我们研究了M的三羧酸循环（TCA）。结核利用遗传学和生物化学的方法相结合，我们已经发现，M。结核病具有一组编码酮戊二酸氧化还原酶的基因，酮戊二酸氧化还原酶是通常在厌氧菌中观察到的TCA的关键酶。此外，我们还发现，编码NADH脱氢酶II（ndh）的基因突变可赋予对异烟肼和异烟酰胺的耐药性。通过结合代谢物分析，稳定同位素通量分析，与突变研究，我们打算表征持久状态的M。结核这些知识应该会导致新的策略，以有效地杀死持续M。结核细胞 公共卫生相关性：迫切需要新的化疗药物来控制结核病大流行，这是由多药和广泛耐药结核分枝杆菌菌株的出现以及该细菌与艾滋病毒的灾难性联盟所推动的。除了对药物的遗传抗性，M.结核病细胞可以成为持续存在的细胞，这些细胞在表型上对药物的杀伤具有抗性。这一建议旨在发展新的理解坚持M。结核细胞为新型和缩短的化疗提供了知识。