Metabolic Impact of Tryptophan Synthase Inhibition in Mycobacterium tuberculosis and Implications for Rational Combination Therapy

项目摘要 凯尔·普朗克是纽约市威尔康奈尔大学医学院药理学项目的博士生， 他是李承晚研究实验室的成员。他的研究兴趣包括开发更深层次的 了解病原微生物中的生物化学过程，并利用这些过程 知识，以制定临床相关的治疗方法，他们引起的感染。在威尔康奈尔大学，凯尔 特别是在结核病（TB）的背景下，这是世界上导致死亡的主要原因， 传染病该奖学金申请详细说明了一项研究计划，其中包括收购 技术技能与导师和职业指导，以准备他的职业目标，建立 一个学术医学中心的独立研究实验室。 凯尔在生物学、生物化学和药理学方面有着丰富的研究经验， 他的研究生工作的基础和这个建议，它集成了系统级的“组学”分析方法 用经典的微生物和生化技术。这个项目的目标是表征代谢 抑制色氨酸生物合成途径的作用，这对分枝杆菌的生存至关重要 结核病（Mtb）-结核病的病原体-并且正被追求为潜在药物靶点的来源。的 该项目有两个目标：确定色氨酸的变构化学抑制剂引起的杀伤机制 合成酶（TrpAB），我们的初步数据表明它比简单的酶产物更复杂 消耗，并调查TrpAB抑制是否会导致其他代谢途径中的附带脆弱性 也可能是有针对性的。通过进行这些调查，凯尔希望为 这些候选药物的作用机制，揭示了色氨酸生物合成的过程及其 Mtb的调节，并揭示了联合治疗的新可能性，这是有效治疗所必需的。 治疗肺结核。 本研究策略建议通过结合标准微生物学技术来回答这些问题 例如生长曲线和菌落形成单位计数， 可诱导的遗传敲减菌株、RNA测序、基于靶向和非靶向LC/MS的代谢组学， 和CRISPR干扰以探测与无药物相比在TrpAB抑制的背景下Mtb的生物学 对照系统水平的组学模式，如代谢组学和转录组学将用于调查 控制Mtb中色氨酸生物合成的遗传和代谢网络，然后将用 遗传和生物化学方法。

PROJECT SUMMARY Kyle Planck is a PhD Student in the pharmacology program at Weill Cornell Medicine in New York City, where he is a member of the Rhee research laboratory. His research interests include developing a deeper understanding of the biochemical processes at work in pathogenic microorganisms and employing this knowledge to formulate clinically relevant treatments for the infections they cause. At Weill Cornell, Kyle carries out this work specifically within the context of tuberculosis (TB), which is the world’s leading cause of death from an infectious disease. This fellowship application details a research plan that incorporates the acquisition of technical skills with mentorship and career guidance in order to prepare him for his career goal of establishing an independent research laboratory at an academic medical center. Kyle has extensive research experience in biology, biochemistry, and pharmacology, and he brings that research foundation to his graduate work and this proposal, which integrates systems level “omics” analytical approaches with classical microbiological and biochemical techniques. The goal of this project is to characterize the metabolic effects of inhibiting the tryptophan biosynthesis pathway, which is essential to the survival of Mycobacterium tuberculosis (Mtb)—the causative agent of TB—and is being pursued as a source of potential drug targets. The project has two aims: to determine the mechanism of killing elicited by allosteric chemical inhibitors of tryptophan synthase (TrpAB), which our preliminary data suggests to be more complex than simple enzyme product depletion, and to survey whether TrpAB inhibition results in collateral vulnerabilities in other metabolic pathways that may be targetable as well. By carrying out these investigations, Kyle hopes to provide evidence for the mechanism of action of these drug candidates, shed light on the process of tryptophan biosynthesis and its regulation in Mtb, and reveal novel possibilities for combination therapy, which is required for the effective treatment of tuberculosis. This research strategy proposes to answer these questions by incorporating standard microbiological techniques such as growth curves and colony-forming unit enumeration in tandem with cutting-edge technologies such as inducible genetic knockdown strains, RNA sequencing, targeted and untargeted LC/MS-based metabolomics, and CRISPR interference to probe the biology of Mtb in the context of TrpAB inhibition compared to no drug controls. Systems level omics modalities such as metabolomics and transcriptomics will be used to survey the genetic and metabolic networks controlling tryptophan biosynthesis in Mtb, which will then be validated with genetic and biochemical approaches.