Chemical probes of mycobacterial growth and persistence

分枝杆菌生长和持久性的化学探针

• 批准号: 9760965
• 负责人: Michael G Wuo
• 金额: $ 6.12万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-20 至 2022-05-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9760965
• 关键词: Antibiotics; Antigens; Antimycobacterial Agents; Bacillus (bacterium); Bacteria; Benign; Biology; Cell Division Process; Cell Wall; Cell division; Cells; Chemicals; Consumption; Corynebacterium; Corynebacterium glutamicum; Data; Diagnosis; Diagnostic; Disease; Drug resistance; Early Diagnosis; Etiology; Fatty Acids; Fluorescence; Fluorescence-Activated Cell Sorting; Foundations; Genetic Determinism; Genomics; Genus Mycobacterium; Growth; Heterogeneity; Human; Hydrolysis; Incidence; Kinetics; Label; Link; Lipids; Metabolic; Methods; Microbe; Microscopy; Modeling; Modification; Molecular; Monitor; Multidrug-Resistant Tuberculosis; Mycobacterium smegmatis; Mycobacterium tuberculosis; Mycolic Acid; Phenotype; Physiology; Population; Population Heterogeneity; Relapse; Reporting; Resistance; Route; Stress; Structure; Time; Trehalose; Tuberculosis; Variant; arabinogalactan; base; cell envelope; cell growth; cost; design; experimental study; flexibility; fluorophore; improved; infectious disease treatment; insight; microbial community; mortality; mycobacterial; mycolate; novel diagnostics; population based; time use; tool; transcriptome sequencing; transcriptomics

PROJECT SUMMARY/ABSTRACT The bacillus Mycobacterium tuberculosis (Mtb) is the etiological agent of tuberculosis (TB) in humans. Despite a steady decline in TB mortality rates, multidrug resistant TB (MDR-TB) and relapse cases have increased. These observations highlight the urgent yet unmet need to understand how MDR-TB develops and persists. Moreover, new strategies to diagnose and treat Mtb infection can limit the spread of the disease and therefore the spread of resistance. Despite these needs, the mechanisms by which Mtb evades treatment remain insufficiently defined, in part due to a lack of tools available to probe Mtb physiology. Strategies that could mitigate these problems include early diagnosis, an understanding of variation in division rate between cells, and insight into Mtb persistence. Distinguishing phenotypic heterogeneity is difficult using traditional transcriptomics; chemical biology can yield tools to detect mycobacteria and monitor the dynamics of cell division. We have developed a fluorogenic probe (QTF) that reports on mycobacterial cell envelope assembly in real time. I shall employ QTF to identify and characterize distinct spatial and temporal profiles of bacteria from the Corynebacterineae suborder. Because QTF is activated by mycolyltransferase activity, fluorescence kinetics will be used for diagnostic purposes to identify and separate mycobacteria and corynebacteria in mixed microbial communities. I will also evaluate QTF in an isogenic monoculture, as I postulate this probe can be used to separate normal from persister cells. Finally, I shall develop a new fluorogenic probe, complementary to QTF, that will covalently label the growing cell wall. I envision QTF and this probe can simultaneously monitor mycolyltransferase activity cell envelope construction and remodeling.

项目概要/摘要 结核分枝杆菌 (Mtb) 是人类结核病 (TB) 的病原体。尽管 结核病死亡率稳步下降，但耐多药结核病（MDR-TB）和复发病例有所增加。这些 观察结果凸显了了解耐多药结核病如何发展和持续的迫切但尚未得到满足的需求。而且， 诊断和治疗结核分枝杆菌感染的新策略可以限制疾病的传播，从而限制结核病的传播 反抗。尽管有这些需求，结核分枝杆菌逃避治疗的机制仍未得到充分明确， 部分原因是缺乏可用于探测结核分枝杆菌生理学的工具。可以缓解这些问题的策略包括 早期诊断、了解细胞间分裂率的变化以及了解结核分枝杆菌的持久性。 使用传统转录组学很难区分表型异质性；化学生物学可以产生 检测分枝杆菌和监测细胞分裂动态的工具。我们开发了荧光探针 （QTF）实时报告分枝杆菌细胞包膜组装。我将使用 QTF 来识别和 表征棒杆菌亚目细菌的不同空间和时间特征。因为QTF 被分枝菌基转移酶活性激活，荧光动力学将用于诊断目的来识别 并分离混合微生物群落中的分枝杆菌和棒状杆菌。我还将评估 QTF 同基因单一培养，因为我假设该探针可用于分离正常细胞和持续细胞。最后，我将 开发一种新的荧光探针，与 QTF 互补，共价标记生长的细胞壁。我 设想 QTF 和该探针可以同时监测霉基转移酶活性细胞包膜结构 和改造。