Study of M. tuberculosis under human host selection to identify virulence and barrier lipids (Project 1)

研究人类宿主选择下的结核分枝杆菌以确定毒力和屏障脂质（项目 1）

• 批准号: 10271484
• 负责人: DAVID Branch MOODY
• 金额: $ 26.04万
• 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/10271484
• 关键词: Bacterial Genes; Biochemical; Biological; Biological Process; CRISPR interference; Cell Wall; Chemicals; Citric Acid Cycle; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Color; Communities; Data; Data Set; Databases; Diagnosis; Diagnostic; Drug Controls; Drug resistance; Epidemic; Gene Silencing; Genes; Genetic; Genetic Variation; Genetic study; Genomics; Genus Mycobacterium; Goals; High Pressure Liquid Chromatography; Human; Immune response; Immunotherapeutic agent; Individual; Infection; Knowledge; Laboratories; Link; Lipids; Maps; Mass Spectrum Analysis; Membrane; Membrane Lipids; Metabolic; Metabolism; Mus; Mycobacterium tuberculosis; Names; Nature; Organism; Outcome; Pathologic; Pathway interactions; Patients; Pattern; Penetration; Pharmaceutical Preparations; Phenotype; Population; Research Personnel; Robin bird; Role; South Africa; Structure; Testing; Tuberculosis; Validation; Variant; Virulence; Virulent; Whole Organism; Wood material; base; cell envelope; comparative; comparative genomics; experimental study; gene discovery; gene function; gene synthesis; genome sequencing; genome wide association study; genome-wide; in vivo; individual patient; insight; lipid biosynthesis; lipidome; lipidomics; metabolomics; mycobacterial; mycolate; novel diagnostics; novel strategies; novel therapeutics; overexpression; pressure; programs; selective expression; whole genome

Project 1. Study of M. tuberculosis under human host selection to identify virulence and barrier lipids Project Leader: D. Branch Moody Coinvestigators: Kyu Rhee, Jacob Mayfield Collaborating Investigators: Adriaan Minnaard (Core C), Jeremy Rock (Core D), Clare Smith (Core E) ABSTRACT Comparative genomics has served as a dominant paradigm for tracking the tuberculosis (TB) epidemic, understanding Mycobacterium tuberculosis (Mtb) virulence and developing new drugs and diagnostics. Mycobacterial metabolism, in contrast, has been viewed as invariant feature of all clinical Mtb strains. Through comparative metabolomic profiling of ~10,000 lipids among 84 patient-derived Mtb strains, we discovered that Mtb’s pathognomonic lipid envelope shows identifiable patterns of variance among strains circulating among human populations. To determine the impact of phenotypic diversity within the infecting bacterial population, we will map cell wall lipid variation among 140 Mtb strains among TB patients from Masiphulemele, South Africa. The resulting lipid map will describe variations in lipid composition among Mtb strains transmitting in community. From a biological perspective, Mtb’s lipid envelope forms the primary interface with the host and is therefore a direct and ongoing biochemical target of evolutionary selection. This project aims to reveal the previously undescribed chemical diversity and lipid products that have arisen as a consequence of host- and drug-derived clinical pressure. Using organism wide lipid profiling and genome wide sequencing, we have identified 42 lipid-gene pairs that dominate in Mtb strain variance, as well as 1150 lipid species overexpressed in virulent Mtb and 250 lipids selectively expressed at the host interface. Preliminary data support our ability to then link these lipids to specific bacterial genes, even when prior to knowledge of the metabolite’s structure or a gene’s function is lacking. CRISPR interference strategies will then establish causal linkages between genes of unknown function and newly discovered lipids. We will further test lipid deficient strains in collaborative cross mice to reveal specific roles of newly identified lipids in Mtb virulence. These discovery studies will identify biologically important lipids that determine key outcomes in virulence, the host interface and Mtb survival in vivo, supporting new approaches for tuberculosis diagnosis and treatment.

项目1。研究M.人类宿主选择下的结核病，以确定毒力和屏障脂质 项目负责人：D.分支穆迪 共同研究者：Kyu Rhee、Jacob Mayfield 合作研究者：Adriaan Minnaard（核心C）、Jeremy Rock（核心D）、克莱尔史密斯（核心E） 摘要 比较基因组学已成为追踪结核病流行的主要范例， 了解结核分枝杆菌（Mtb）的毒力，开发新的药物和诊断方法。 相比之下，分枝杆菌代谢被视为所有临床Mtb菌株的不变特征。通过 通过对84个患者来源的结核分枝杆菌菌株中约10，000种脂质的比较代谢组学分析，我们发现， 结核分枝杆菌的特异性脂质包膜显示出可识别的变异模式， 人类种群。为了确定感染细菌群体内表型多样性的影响， 我们将绘制来自南非Masiphulemele的结核病患者中140株结核分枝杆菌菌株的细胞壁脂质变异图， 非洲由此产生的脂质图谱将描述结核分枝杆菌菌株中脂质组成的变化， 社区从生物学的角度来看，结核分枝杆菌的脂质包膜形成了与宿主的主要界面， 因此是进化选择的直接和持续的生化目标。该项目旨在揭示 以前未描述的化学多样性和脂质产品，已出现的结果，主机-和 药物引起的临床压力。使用生物体范围的脂质分析和基因组范围的测序，我们 鉴定了在Mtb菌株变异中占主导地位的42个脂质基因对，以及1150个过表达的脂质种类 在毒性Mtb和250脂质选择性地表达在宿主界面。初步数据支持我们的能力， 然后将这些脂质与特定的细菌基因联系起来，即使在知道代谢物的结构之前， 基因的功能缺失。然后，CRISPR干扰策略将在基因之间建立因果联系。 功能未知和新发现的脂质。我们将在合作杂交中进一步测试脂质缺乏菌株 小鼠揭示新鉴定的脂质在结核分枝杆菌毒力中的特定作用。这些发现研究将确定 生物学上重要的脂质，决定了毒力，宿主界面和Mtb存活的关键结果， 体内，支持结核病诊断和治疗的新方法。