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

色氨酸合酶抑制对结核分枝杆菌的代谢影响以及合理联合治疗的意义

• 批准号: 10442387
• 负责人: Kyle Planck
• 金额: $ 4.68万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10442387
• 关键词: Academic Medical Centers; Anabolism; Animal Model; Antibiotic Therapy; Antibiotics; Antimycobacterial Agents; Antitubercular Agents; Biochemical; Biochemical Pathway; Biochemical Process; Biochemistry; Biology; CRISPR interference; Cause of Death; Cells; Chemicals; Colony-forming units; Combined Modality Therapy; Communicable Diseases; Complex; Data; Development; Diet; Dominant-Negative Mutation; Drug Combinations; Drug Controls; Drug Targeting; Drug resistance; Enzymes; Extreme drug resistant tuberculosis; Feedback; Fellowship; Folic Acid; Foundations; Genes; Genetic; Goals; Growth; Human; Infection; Investigation; Iron; Knock-out; Knowledge; Laboratory Research; Light; Link; Literature; Medicine; Mentorship; Metabolic; Metabolic Pathway; Microbiological Techniques; Microbiology; Modality; Molecular Target; Mycobacterium tuberculosis; New York City; Nutrient; Organism; Pathogenesis; Pathogenicity; Pathway interactions; Persons; Pharmacology; Physiology; Population; Process; Production; Regulation; Research; Resistance; Siderophores; Signal Transduction; Source; Starvation; Stress; Supplementation; Surveys; System; Technical Expertise; Techniques; Technology; Testing; Toxic effect; Tryptophan; Tryptophan Synthase; Tuberculosis; Vitamin K 2; Vocational Guidance; Western Blotting; Work; base; career; chemotherapy; clinically relevant; cost; doctoral student; drug action; drug candidate; drug development; effective therapy; experience; fitness; gene product; inhibitor; interest; knock-down; liquid chromatography mass spectrometry; member; metabolomics; microorganism; mycobacterial; novel; novel strategies; pathogen; programs; protein expression; resistant strain; response; scaffold; transcriptome sequencing; transcriptomics; treatment strategy; tuberculosis drugs; tuberculosis treatment

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.

项目总结 凯尔·普朗克是纽约市威尔·康奈尔医学院药理学项目的博士生。 他是Rhee研究实验室的成员。他的研究兴趣包括开发更深层次的 理解病原微生物的生物化学过程，并利用这一点 为他们引起的感染制定临床相关治疗方法的知识。在威尔·康奈尔大学，凯尔携带 这项工作特别是在结核病(TB)的背景下进行的，结核病是世界上主要的死亡原因 一种传染病。这份奖学金申请详细说明了一项研究计划，该计划包括收购 具有指导和职业指导的技能，以便为他的职业目标做好准备 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丰富的研究经验，他带来了这些研究 为他的研究生工作奠定了基础，并提出了这一建议，其中整合了系统级的“组学”分析方法 用经典的微生物和生化技术。这个项目的目标是描述新陈代谢的特征 抑制对分枝杆菌生存至关重要的色氨酸生物合成途径的作用 结核病(Mtb)--结核病的病原体--正在作为潜在药物靶标的来源而被追查。这个 该项目有两个目标：确定色氨酸变构化学抑制剂诱导的杀伤机制 合成酶(TrpAB)，我们的初步数据表明它比简单的酶产物更复杂 耗竭，并调查TrpAB抑制是否会导致其他代谢途径的侧支脆弱性 这也可能是有针对性的。通过开展这些调查，凯尔希望为 这些候选药物的作用机制，有助于揭示色氨酸的生物合成过程及其 对结核分枝杆菌的调节，并揭示了有效联合治疗的新可能性 结核病的治疗。 这一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与尖端技术相结合，例如 可诱导基因敲除菌株，RNA测序，靶向和非靶向LC/MS代谢组学， 以及CRISPR干扰以探测TrpAB抑制背景下与无药物相比的Mtb的生物学 控制。将使用代谢组学和转录组学等系统水平组学方法来调查 控制Mtb中色氨酸生物合成的遗传和代谢网络，然后将通过 遗传和生化方法。