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: 10438911
• 负责人: DAVID Branch MOODY
• 金额: $ 257.81万
• 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/10438911
• 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; diagnostic strategy; 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.

摘要 - 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，这在很大程度上是由于 其细胞膜的极其不寻常的生物和化学特性。由具有特色的 疏水性分枝菌外膜，固定在多糖的底层复合物上 肽聚糖聚合物，结核分枝杆菌包膜既是与人类的主要界面，也是人类的屏障 主持人。在人类结核 (TB) 疾病中，结核分枝杆菌包膜介导长达数年的对峙，并充当 对所有抗分枝杆菌药物具有屏障作用。然而，对其天然组成、变异和调节的了解 毒品进入仍然零碎。该申请人团队创建了新的遗传和代谢组学工具 全面剖析和分析 Mtb 包膜的代谢物和脂质成分 跨大量临床分离株的生物体范围的基础。此外，该 TBRU 提议提供第一个 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 毒力和药物作用障碍可以为开发更好的诊断和治疗提供信息。 新分子的结构将首先使用合成化学和质谱法确定。这 然后将使用新的基因组规模来识别编码这些代谢物的基因并进行功能验证 CRISPR 干扰技术、渗透细胞膜的分析以及基因定义 体内生长的小鼠模型。使用质谱法，我们将解析多达 250 个表面的结构 屏障脂质和超过 41 个基因-脂质对在患者细胞包膜变异中占主导地位。 患者来源的 Mtb 菌株将从我们位于 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来检测呼出气中的活结核分枝杆菌（非 咳嗽）人类生物气溶胶。屏障功能的研究特别强调利福平作为模型 化合物由于其作为一线药物的临床重要性以及作为耐药结核病的决定因素的作用。 分析患者尿液和血清的能力进一步发现了新的生物标志物 疾病活动和对药物治疗的反应，激发相关的转化努力以推进 开发基于非痰的实时护理点诊断测试。这个互动性很强的团体 因此，科学家们寻求提供更好的药物和诊断测试，以及深入而持久的科学依据。 为了解 Mtb 包膜，特别是控制 Mtb 的特定基因和分子奠定基础 主动重塑、药物作用和人类宿主反应。