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

基本信息

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
项目类别：
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

项目摘要

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）：迫切需要新的化学治疗剂来控制结核病大流行，这是由于多药和广泛耐药的结核病菌株以及细菌与HIV的灾难性联盟的出现所助长的。除了对药物的遗传性抗性外，结核分枝杆菌细胞还可以成为持久性，在表型上对药物杀死具有抗性。进入持续状态的能力需要长期的化学疗法，这代表了改善TB化学疗法的最大挑战。不幸的是，对持久者的代谢状态知之甚少。为了阐明持续性结核分枝杆菌细胞的生理状态，我们已经使用质谱法在有氧和低氧生长条件下生长（持久性模型）时，可以产生结核分枝杆菌的代谢特征，并发现了几个关键控制点调节NADH/NADH/NADH/NADH/NADH/NADH/NADH（氧化还原））代谢。为了进一步表征代谢谱，我们研究了结核分枝杆菌的三羧酸周期（TCA）。使用遗传学和生化方法的结合，我们发现结核分枝杆菌具有编码酮戊二酸氧化还原酶的一组基因，这是TCA的关键酶，这是厌氧菌中通常观察到的。此外，我们发现编码NADH脱氢酶II（NDH）会议的基因中的突变对异oni氮和乙二酰胺的抗性。通过将代谢物分析和稳定的同位素通量分析与突变研究相结合，我们打算表征结核分枝杆菌的持续状态。这些知识应导致新的策略有效地杀死持续的结核分枝杆菌细胞。公共卫生相关性：迫切需要新的化学治疗剂来控制结核病大流行，这是由于多药和广泛的药物耐药性结核菌菌株的出现以及细菌与艾滋病毒的灾难性联盟所促进的。除了对药物的遗传性抗性外，结核分枝杆菌细胞还可以成为持久性，在表型上对药物杀死具有抗性。该建议旨在发展对持续的结核分枝杆菌细胞的新理解，从而为新颖和缩短化学疗法提供知识。