Identification of new inhibitors of essential functions in M. tuberculosis by high-throughput metabolic profiling

通过高通量代谢分析鉴定结核分枝杆菌基本功能的新抑制剂

• 批准号: 10568482
• 负责人: Michael Berney
• 金额: $ 65.38万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-18 至 2027-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10568482
• 关键词: Algorithms; Anti-Bacterial Agents; Antibiotics; Antineoplastic Agents; Antitubercular Agents; Atlases; Biochemical; Biological; Biological Assay; Biology; CRISPR interference; CRISPR/Cas technology; Cancer cell line; Categories; Cells; Chemicals; Communicable Diseases; Computer Models; Consumption; Custom; Data; Databases; Dimensions; Droughts; Drug Kinetics; Drug Screening; Drug Targeting; Ensure; Environment; Escherichia coli; Essential Genes; Event; Expression Library; Fingerprint; Genes; Genetic; Goals; Growth; Growth Inhibitors; Lead; Libraries; Mass Spectrum Analysis; Measures; Metabolic; Methodology; Methods; Molecular; Multidrug-Resistant Tuberculosis; Mycobacterium smegmatis; Mycobacterium tuberculosis; Nature; Pharmaceutical Preparations; Phenotype; Property; Proteolysis; Regulator Genes; Reproducibility; Safety; Switzerland; System; Techniques; Technology; Time; Validation; Work; antimicrobial; comparative; computerized tools; drug action; drug candidate; drug discovery; emerging antibiotic resistance; follow-up; gene function; in vivo; inhibitor; innovation; insight; knock-down; lung cancer cell; metabolic profile; metabolome; metabolomics; meter; novel; novel strategies; overexpression; response; small molecule; small molecule libraries; transcription factor; tuberculosis drugs

Project Summary/Abstract While the emergence of multi-drug resistant tuberculosis raises an urgent need for antimicrobials with new Modes of Action, their discovery remains a major challenge. Many of the techniques to unravel drug Modes of Action rely on low-throughput, time-consuming and target-specific approaches that provide low- dimensional views into the broader functional impact of potential drugs. Together with the Zampieri lab at ETH Zurich, Switzerland, by leveraging CRISPR technology and non-targeted metabolomics, we developed a combined computational/experimental strategy that is based on the comparison of genetic and drug induced metabolic effects and allows to perform high-throughput de novo functional annotations of large compound libraries. Unraveling the mechanistic basis of drug or gene perturbations of thousands of metabolites provides rich multidimensional information complementary to classical phenotypic profiling and can be used to investigate the effect and mode of action of any drug candidate. The overall goal of the present proposal is to achieve functional annotation of 500 anti-TB compounds with known potency but unknown MoA, which will pave the way to new unconventional strategies to eradicate TB. We will build a compendium of metabolic responses of Mtb to essential gene knockdown, transcription factor overexpression and a unique selection of libraries of Mtb growth inhibitors. We will use our custom- developed computational tools to categorize drug action and gene knockdowns according to metabolic profiles, make testable hypothesis about unconventional drug MoA and move prioritized compounds to genetic and biochemical hit validation. An important collateral benefit of our proposed work will be functional annotation of genes with yet unknown function in M. tuberculosis, and an information dense database on gene-drug-metabolic interactions in M. tuberculosis.

项目总结/摘要 虽然耐多药结核病的出现提出了对具有新的耐药性的抗微生物剂的迫切需求， 虽然它们的作用方式不同，但它们的发现仍然是一个重大挑战。许多破解毒品模式的技术 行动依赖于低通量、耗时和针对具体目标的方法， 从多个维度观察潜在药物的更广泛的功能影响。与Zampieri实验室一起， 瑞士苏黎世ETH，通过利用CRISPR技术和非靶向代谢组学， 开发了一种结合计算/实验的策略，该策略基于遗传学的比较， 和药物诱导的代谢作用，并允许进行高通量从头功能注释 大型复合图书馆。揭开数千种药物或基因干扰的机制基础 代谢物的分析提供了丰富的多维信息，与经典的表型分析互补 并可用于研究任何候选药物的作用效果和作用模式。 本提案的总体目标是实现500种抗TB化合物的功能注释， 已知的效力，但未知的MoA，这将为新的非传统战略铺平道路，以消除 TB.我们将建立一个结核分枝杆菌代谢反应的纲要，以必要的基因敲除，转录 因子过表达和Mtb生长抑制剂文库的独特选择。我们会用我们的习惯- 开发了计算工具，根据代谢分类药物作用和基因敲除 简介，使非常规药物MoA的可测试的假设和移动优先化合物， 基因和生化命中验证。我们拟议工作的一个重要附带好处是 对M.结核病和信息密集 基因-药物-代谢相互作用的数据库。结核