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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要凯尔·普朗克 (Kyle Planck) 是纽约市威尔康奈尔医学院药理学项目的博士生，该校他是李研究实验室的成员，他的研究兴趣包括开发更深入的研究。了解病原微生物的生化过程并加以利用在威尔康奈尔大学，凯尔拥有针对其引起的感染制定临床相关治疗方法的知识。特别是在结核病 (TB) 的背景下开展这项工作，结核病是世界上导致死亡的主要原因该奖学金申请详细介绍了一项研究计划，其中包括收购技术技能以及导师和职业指导，以便为他建立职业目标做好准备学术医疗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拥有丰富的研究经验，他带来了这些研究为他的研究生工作和该提案奠定了基础，该提案集成了系统级“组学”分析方法该项目的目标是利用经典的微生物学和生化技术来表征代谢。抑制色氨酸生物合成途径的作用，色氨酸生物合成途径对分枝杆菌的生存至关重要结核病 (Mtb)——结核病的病原体——正被视为潜在药物靶点的来源。该项目有两个目标：确定色氨酸变构化学抑制剂引起的杀伤机制合酶 (TrpAB)，我们的初步数据表明它比简单的酶产品更复杂耗竭，并调查 TrpAB 抑制是否会导致其他代谢途径的附带脆弱性凯尔希望通过进行这些调查来提供证据。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结核分枝杆菌的调节，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结核病。该研究策略建议通过结合标准微生物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的结合，例如可诱导的基因敲低菌株、RNA 测序、基于 LC/MS 的靶向和非靶向代谢组学、与无药物相比，在 TrpAB 抑制的背景下使用 CRISPR 干扰来探究 Mtb 的生物学系统级组学模式（例如代谢组学和转录组学）将用于调查控制结核分枝杆菌色氨酸生物合成的遗传和代谢网络，然后将通过以下方法进行验证遗传和生化方法。