Metabolic determinants of Mtb virulence, vulnerability and variation

结核分枝杆菌毒力、脆弱性和变异的代谢决定因素

• 批准号: 10271478
• 负责人: DAVID Branch MOODY
• 金额: $ 254.73万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271478
• 关键词: Advanced Development; Aerosols; Anabolism; Antibiotics; Bacteria; Biological; Biological Assay; Biological Markers; Biology; Blood; CRISPR interference; Characteristics; Chemicals; Clinical; Complex; DNA-Directed RNA Polymerase; Detection; Development; Diagnostic; Diagnostic tests; Disease; Drug Regulations; Drug resistance in tuberculosis; Elements; Ethambutol; Exhalation; Foundations; Genes; Genetic; Genomics; Genus Mycobacterium; Growth; Human; Hydrophobicity; Immune response; Knowledge; Lecithin; Link; Lipids; Maps; Mass Spectrum Analysis; Mediating; Membrane; Membrane Lipids; Metabolic; Modeling; Mus; Mycobacterium tuberculosis; Organism; Pathologic; Patients; Penetration; Peptidoglycan; Performance; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Physiology; Plasma; Polymers; Polysaccharides; Prevalence; Rapid diagnostics; Research; Resistance; Rifampin; Role; Sampling; Scientist; Serum; Site; South Africa; Species Specificity; Sphingomyelins; Sputum; Structure; Surface; Synthesis Chemistry; System; Technology; Therapeutic; Time; Tuberculosis; Urine; Variant; Virulence; Virulent; Whole Organism; active control; arabinofuranose; base; cell envelope; chemical property; clinically relevant; comparative; drug action; drug testing; drug-sensitive; gene synthesis; genome wide association study; genome-wide; in vivo Model; interdisciplinary approach; lipid biosynthesis; lipidomics; metabolic profile; metabolomics; mouse model; mycobacterial; mycolate; overexpression; pandemic disease; pathogen; patient population; point-of-care diagnostics; response; response biomarker; tool; treatment response; urinary

ABSTRACT - Metabolic determinants of Mtb virulence, vulnerability and variation Mycobacterium tuberculosis (Mtb) has emerged as the world's most deadly pathogen based in large part on the highly unusual biological and chemical properties of its cell envelope. Comprised of a distinctive hydrophobic outer mycolate membrane, anchored to an underlying complex of polysaccharide and peptidoglycan polymers, the Mtb envelope serves as both the primary interface with, and barrier to, the human host. In human tuberculosis (TB) disease the Mtb envelope mediates a years-long standoff, and serves as the barrier to all anti-mycobacterial drugs. Yet, knowledge of its native composition, variation and regulation of drug entry remains fragmentary. This team of applicants has created new genetic and metabolomic tools to comprehensively dissect and analyze the metabolite and lipid components of the Mtb envelope on an organism-wide basis across a large set of clinical isolates. Moreover, this TBRU proposes to provide the first descriptions of cell envelope variation among isolates from human patients and identify key determinants of its virulence and barrier to drug action that could inform the development of better diagnostics and therapeutics. Structures of new molecules will first be determined using synthetic chemistry and mass spectrometry. The genes encoding these metabolites will then be identified and functionally validated using new genome-scale CRISPR interference technologies, assays for penetration into the cell envelope, and genetically defined mouse models of in vivo growth. Using mass spectrometry, we will solve the structures of up to 250 surface barrier lipids and more than 41 gene-lipid pairs that dominate in cell envelope variation among patients. Patient-derived Mtb strains will be obtained from clinical samples collected at our field sites in Masiphumelele, South Africa, where we will implement clinically relevant technology for detection of live Mtb in exhaled (non- coughed) human bioaerosols. Studies of barrier function place special emphasis on rifampicin as a model compound due to its clinical importance as a frontline drug and role as a defining element of drug resistant TB. The ability to analyze patient urine and serum has further resulted in the discovery of new biomarkers of disease activity and response to drug therapy, motivating linked translational efforts to advance the development of non-sputum based, real time point-of-care diagnostic tests. This highly interactive group of scientists thus seeks to provide better drugs and diagnostic tests, as well as a deep and durable scientific foundation for understanding of the Mtb envelope, especially the particular genes and molecules that control active remodeling, drug action and human host response.

结核分枝杆菌毒力、脆弱性和变异的代谢决定因素 结核分枝杆菌（Mtb）已成为世界上最致命的病原体，这在很大程度上是基于 它的细胞被膜具有极不寻常的生物和化学特性。由一种独特的 疏水性的真菌酸盐外膜，锚定在下面的多糖复合物上， 作为肽聚糖聚合物，Mtb包膜既作为与人的主要界面，又作为人的屏障。 主持人在人类结核病（TB）疾病中，Mtb包膜介导了长达数年的对峙，并充当了 所有抗分枝杆菌药物的屏障。然而，了解其天然成分，变异和调节， 药物输入仍然不完整。这个申请人团队创造了新的遗传和代谢组学工具， 全面解剖和分析结核分枝杆菌包膜的代谢产物和脂质成分， 在大量临床分离株中的微生物范围内的基础。此外，该TBRU建议提供第一个 描述人类患者分离株之间的细胞包膜变异，并确定其关键决定因素 毒力和药物作用的障碍，可以为更好的诊断和治疗的发展提供信息。 新分子的结构将首先使用合成化学和质谱法确定。的 编码这些代谢物的基因将被识别，并使用新的基因组规模进行功能验证。 CRISPR干扰技术，渗透到细胞包膜的测定，以及基因定义的 体内生长的小鼠模型。使用质谱分析，我们将解决多达250个表面的结构 屏障脂质和超过41个基因-脂质对在患者的细胞包膜变化中占主导地位。 患者来源的结核分枝杆菌菌株将从我们在Masiphumelele的现场采集的临床样本中获得， 南非，我们将在那里实施临床相关技术，用于检测呼出（非 咳嗽）人体生物气溶胶。屏障功能的研究特别强调以利福平为模型 由于其作为一线药物的临床重要性和作为耐药结核病的决定性因素的作用， 分析患者尿液和血清的能力进一步导致了新的生物标志物的发现， 疾病活动和对药物治疗的反应，激励相关的翻译努力，以促进 开发基于非痰液的、真实的时间点护理诊断测试。这个高度互动的群体， 因此，科学家们寻求提供更好的药物和诊断测试，以及一个深入和持久的科学， 这是理解结核分枝杆菌包膜的基础，特别是控制结核分枝杆菌包膜的特定基因和分子。 主动重塑、药物作用和人类宿主反应。