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: 10612023
• 负责人: DAVID Branch MOODY
• 金额: $ 256.65万
• 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/10612023
• 关键词: Advanced Development; Aerosols; Anabolism; Antibiotics; Antimycobacterial Agents; 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; 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; 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 - MTB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，很大程度上是基于 其细胞包膜的高度不寻常的生物学和化学特性。由独特的 疏水外乳腺膜膜，固定在多糖的基础络合物上 肽聚糖聚合物，MTB信封既是与人类的主要界面，又是障碍 主持人。在人类结核病（TB）疾病中，MTB包膜介导了长达数年的僵局，并充当 所有抗菌药物的障碍。然而，了解其本地组成，变化和调节的知识 药物进入仍然是零散的。这个申请人团队创建了新的遗传和代谢组工具 全面剖析和分析MTB信封的代谢物和脂质成分 在大量临床分离株中范围内的有机体基础。而且，该TBRU建议提供第一个 人类患者分离株之间细胞包膜变化的描述，并确定其关键决定因素 毒品作用的毒力和障碍，可以为更好的诊断和治疗学的发展提供依据。 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产物的基因并在功能上验证 CRISPR干扰技术，渗透到细胞包膜中的测定以及遗传定义 体内生长的小鼠模型。使用质谱法，我们将解决高达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。 将从我们在Masiphumelele的现场收集的临床样本中获得患者来源的MTB菌株， 南非，我们将实施与临床相关的技术来检测呼出的现场MTB（非 - 咳嗽）人类生物溶质。屏障功能的研究将利福平作为模型特别强调 化合物由于其作为前线药物的临床重要性和作为耐药性结核病的定义元素而引起的。 分析患者尿液和血清的能力进一步导致了发现的新生物标志物 疾病活动和对药物疗法的反应，激励连接的转化努力以推动 开发基于非输入的实时护理诊断测试。这个高度互动的群体 因此，科学家试图提供更好的药物和诊断测试，以及深层耐用的科学 理解MTB信封的基础，尤其是控制的特定基因和分子 主动重塑，药物作用和人类宿主反应。