Mechanisms of macrolide synergy in Mycobacterium tuberculosis

大环内酯类药物在结核分枝杆菌中的协同作用机制

• 批准号: 10386174
• 负责人: Francesca Guglielmini Tomasi
• 金额: $ 2.16万
• 依托单位: HARVARD SCHOOL OF PUBLIC HEALTH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2022-08-22
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386174
• 关键词: Acetylation; Address; Amino Acyl Transfer RNA; Aminoacyl-tRNA hydrolase; Animal Model; Antibiotics; Bacterial Infections; Binding; Biological Assay; CRISPR interference; Cell Survival; Cells; Charge; Collaborations; Combined Modality Therapy; Communicable Diseases; Coupled; Creativeness; Critical Thinking; Data Analyses; Drops; Drug Metabolic Detoxication; Drug Targeting; Drug resistance in tuberculosis; Enzymes; Exposure to; Fellowship; Genes; Genetic; Goals; Grant; Growth; Human; Ligase; Liquid Chromatography; Macrolides; Mass Spectrum Analysis; Measures; Methods; Morbidity - disease rate; Mycobacterium smegmatis; Mycobacterium tuberculosis; Natural Resistance; Organism; Paper; Pathway interactions; Patients; Peptides; Persons; Pharmaceutical Preparations; Phenotype; Physiological; Play; Prevalence; Principal Investigator; Problem Solving; Protein Biosynthesis; Proteins; Public Health; Public Health Schools; Quality Control; RNA, Transfer, Amino Acid-Specific; Recording of previous events; Recycling; Research; Resistance; Ribosomes; Role; Sampling; System; Technical Expertise; Techniques; Testing; Toxin; Training; Transfer RNA; Transfer RNA Aminoacylation; Translations; Tuberculosis; Writing; acquired drug resistance; antibiotic tolerance; antitoxin; drug discovery; experimental study; fight against; fitness; genetic approach; global health; graduate student; improved; inhibitor; interest; knock-down; mortality; novel strategies; novel therapeutics; overexpression; pathogen; peptidyl-tRNA; premature; response; skills; synergism; tool; tuberculosis drugs; tuberculosis treatment

ABSTRACT I have been fortunate to combine my passions for scientific research and public health in my doctoral studies at the Harvard T. H. Chan School of Public Health. My goal is to combine creativity and technical skills in drug discovery efforts against infectious diseases of high morbidity and mortality. Dr. Eric Rubin's lab is the archetype for rigorous scientific training and creative problem-solving. He and I have established my core training goals. I hope not only to complete my proposed research plan, but also to continue sharpening skills in grant and paper writing, collaboration, critical thinking, and quantitative data analysis. An F31 Fellowship will propel me in my journey to become a principal investigator in the field of antibiotic research and discovery. Mycobacterium tuberculosis (Mtb), which causes tuberculosis (TB), has been a leading cause of infectious disease morbidity and mortality for thousands of years. One major reason for its global persistence is the fact that Mtb is intrinsically resistant to most antibiotics. Current treatment requires months of combination therapy with drugs almost exclusively reserved for TB and that are not well-tolerated by patients. The high morbidity, mortality, and the growing prevalence of acquired drug resistance in TB motivate an urgent search for new drugs. If bacterial targets could be identified that synergize with existing drugs, such as to bring efficacy to antibiotics not currently effective against TB, they could advance our goal of treatment improvement. I have used a genetic approach to identify an essential Mtb enzyme that, when depleted, sensitizes this pathogen to existing antibiotics. Here, I propose to characterize the mechanisms of synergy between this Mtb gene and macrolide antibiotics, which bind the 50S ribosomal subunit to inhibit translation. Macrolides are inexpensive and well- tolerated antibiotics used to treat other infectious diseases. My gene of interest, pth, encodes the essential enzyme peptidyl tRNA hydrolase, a translation rescue factor that cleaves tRNA from peptides when they are prematurely released from stalled ribosomes. My approach to studying Pth as an antibiotic target draws on existing analytical techniques, as well as a new method using tRNA-sequencing that I have developed for studying charged tRNA pools. Investigating synergistic drug targets and their mechanisms of interaction will allow us to lend new efficacy to old compounds in the fight against TB. Additionally, new tools to study tRNA will allow us to expand our understanding of translation machinery in other organisms.

抽象的 我很幸运地将我的热情结合在我的博士学位上 哈佛T. H. Chan公共卫生学院的研究。我的目标是结合创造力和技术技能 在针对高发病率和死亡率的传染病的药物发现工作中。埃里克·鲁宾博士的实验室是 严格的科学培训和创造性问题的原型。他和我已经建立了我的核心培训 目标。我希望不仅能够完成我提出的研究计划，还希望继续提高格兰特的技能 以及纸质写作，协作，批判性思维和定量数据分析。 F31奖学金将推进 我在成为抗生素研究和发现领域的首席研究员的旅程中。 导致结核病（TB）的结核分枝杆菌（MTB）一直是主要原因 传染病发病率和死亡率数千年。全球持久性的主要原因之一是 MTB本质上对大多数抗生素具有抗性。当前的治疗需要数月的组合 药物的治疗几乎仅用于结核病，患者不耐受。高 结核病的发病率，死亡率和越来越多的毒品耐药性促使人们紧急寻找 新药。如果可以确定细菌靶标与现有药物协同作用，例如将功效带入 抗生素目前尚未针对结核病有效，它们可以提高我们改善治疗的目标。我已经用过了 一种遗传方法来识别必需的MTB酶，该酶在耗尽时将这种病原体敏感 抗生素。在这里，我建议表征该MTB基因和大花环之间的协同作用 抗生素，结合50S核糖体亚基以抑制翻译。大环内酯类廉价且良好 耐受的抗生素用于治疗其他传染病。我感兴趣的基因PTH编码了必不可少的 酶肽基TRNA水解酶，这是一种翻译救援因子，在肽中裂解tRNA时。 从停滞的核糖体过早释放。我研究PTH作为抗生素靶标的方法是 现有的分析技术以及我为我开发的tRNA测序的一种新方法 研究带电的tRNA池。研究协同药物靶标及其相互作用机制将 允许我们在与结核病的斗争中对旧化合物提出新的功效。此外，研究tRNA的新工具将 让我们扩大对其他生物中翻译机制的理解。