Mechanisms of macrolide synergy in Mycobacterium tuberculosis

摘要 在我的博士生涯中，我有幸将我对科学研究和公共卫生的热情结合在一起 在哈佛大学公共卫生学院学习。我的目标是将创造力和技术技能结合起来 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 严谨的科学训练和创造性解决问题的原型。他和我已经确立了我的核心训练 目标。我不仅希望完成我提出的研究计划，而且还希望在赠款中继续磨练技能 论文写作、协作、批判性思维和定量数据分析。F31联谊会将推动 我在成为抗生素研究和发现领域的首席研究员的旅程中。 引起结核病(TB)的结核分枝杆菌(Mtb)一直是导致 几千年来传染病的发病率和死亡率。它在全球范围内持续存在的一个主要原因是 事实上，结核分枝杆菌对大多数抗生素具有内在的抗药性。目前的治疗方法需要几个月的联合 使用几乎只针对结核病的药物进行治疗，患者对这些药物的耐受性不佳。高潮 结核病的发病率、死亡率和获得性耐药性的日益流行促使人们迫切地寻找 新药。如果能确定与现有药物协同的细菌靶点，例如将疗效带给 抗生素目前对结核病无效，它们可能会推进我们改进治疗的目标。我用过 一种识别一种必要的结核分枝杆菌酶的遗传学方法，当这种酶耗尽时，会使这种病原体对现有的 抗生素。在这里，我建议描述该mtb基因与大环内酯类化合物之间的协同作用机制。 抗生素，它结合了50s核糖体亚基来抑制翻译。大环内酯价格便宜，而且性能良好- 用于治疗其他传染病的耐受抗生素。我感兴趣的基因PTH编码了 多肽tRNA水解酶，一种翻译救援因子，当多肽被切割时，它能从多肽中切割出tRNA 过早地从停滞的核糖体中释放。我将甲状旁腺素作为抗生素靶标的研究方法借鉴了 现有的分析技术，以及我为之开发的使用tRNA测序的新方法 研究带电的tRNA池。研究协同药物靶点及其相互作用机制将 使我们能够在抗击结核病的斗争中赋予旧化合物新的效力。此外，研究tRNA的新工具将 使我们能够扩大对其他生物体中翻译机制的理解。

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.