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.

摘要 我很幸运地将我对科学研究和公共卫生的热情结合在我的博士学位上。 在哈佛T. H.陈氏公共卫生学院。我的目标是把联合收割机的创造力和技术能力结合起来 在针对高发病率和高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 严格的科学训练和创造性解决问题的典范。他和我建立了我的核心训练 目标.我希望不仅能完成我提出的研究计划，而且能在格兰特继续磨练技能 论文写作，协作，批判性思维和定量数据分析。F31奖学金将推动 在我成为抗生素研究和发现领域的首席研究员的旅程中，我一直在努力。 引起结核病（TB）的结核分枝杆菌（Mtb）一直是导致结核病的主要原因。 传染病的发病率和死亡率几千年。它在全球持续存在的一个主要原因是 结核分枝杆菌对大多数抗生素具有内在耐药性。目前的治疗需要几个月的组合 使用几乎专门用于结核病的药物进行治疗，患者耐受性不佳。高 结核病的发病率、死亡率和获得性耐药性的日益普遍，促使人们迫切寻求 新药如果能够确定与现有药物协同作用的细菌靶点，例如将疗效提高到 抗生素目前对结核病无效，它们可以推进我们改善治疗的目标。我用过 一种基因方法来确定一种必需的结核分枝杆菌酶，当耗尽时，这种酶会使这种病原体对现有的结核分枝杆菌敏感， 抗生素在这里，我建议描述Mtb基因和大环内酯类之间协同作用的机制 抗生素，其结合50 S核糖体亚基以抑制翻译。大环内酯类药物价格低廉，而且- 用于治疗其他传染病的耐受性抗生素。我感兴趣的基因，pth，编码了 酶肽基tRNA水解酶，一种翻译拯救因子，当肽 从停滞的核糖体中过早释放出来。我研究PTH作为抗生素靶点的方法借鉴了 现有的分析技术，以及一种新的方法，使用tRNA测序，我已经开发了 研究带电的tRNA池研究协同药物靶点及其相互作用机制将 使我们能够在对抗结核病的斗争中为旧的化合物提供新的功效。此外，研究tRNA的新工具将 使我们能够扩展对其他生物翻译机制的理解。