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）

基本信息

批准号：
10612035
负责人：
DAVID Branch MOODY
金额：
$ 25.84万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
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/10612035
关键词：
Antimycobacterial Agents Bacterial Genes Biochemical Biological Biological Process CRISPR interference Cell Wall Chemicals Citric Acid Cycle Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Color Communities Data Data Set Databases Diagnostic Drug Controls Drug resistance Epidemic Gene Silencing Genes Genetic Genetic Variation Genetic study Genomics Genus Mycobacterium Goals High Pressure Liquid Chromatography Human Immune Immune response Individual Infection Knowledge Laboratories Link Lipids Maps Mass Spectrum Analysis Membrane Membrane Lipids Metabolic Metabolism Mus Mycobacterium tuberculosis Names Nature Organism Outcome Pathway interactions Patients Pattern Penetration Pharmaceutical Preparations Phenotype Population Research Personnel Robin bird Role South Africa Structure Testing Tuberculosis Tuberculosis diagnosis Validation Variant Virulence Virulent Whole Organism Wood material 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 transmission process tuberculosis treatment whole genome

项目摘要

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. 在人类宿主选择下研究结核分枝杆菌，以确定毒力和屏障脂质项目负责人：D. Branch Moody 联合调查员：Kyu Rhee、雅各布·梅菲尔德合作调查员：Adriaan Minnaard（核心 C）、Jeremy Rock（核心 D）、Clare Smith（核心 E）抽象的比较基因组学已成为追踪结核病（TB）流行的主导范式，了解结核分枝杆菌 (Mtb) 毒力并开发新药和诊断方法。相比之下，分枝杆菌代谢被视为所有临床结核分枝杆菌菌株的不变特征。对 84 种源自患者的 Mtb 菌株中约 10,000 种脂质进行比较代谢组学分析，我们发现结核分枝杆菌的特征性脂质包膜显示出在不同人群中传播的菌株之间可识别的变异模式。确定感染细菌群体内表型多样性的影响，我们将绘制来自南部 Masiphulemele 的结核病患者中 140 个 Mtb 菌株的细胞壁脂质变异图非洲。由此产生的脂质图将描述在非洲传播的结核分枝杆菌菌株之间脂质成分的变化。从生物学角度来看，结核分枝杆菌的脂质包膜形成了与宿主的主要界面。因此，该项目旨在揭示进化选择的直接且持续的生化目标。先前未描述的化学多样性和脂质产物是由于宿主和利用全生物体脂质分析和全基因组测序，我们获得了药物衍生的临床压力。鉴定了 42 个在 Mtb 菌株变异中占主导地位的脂质基因对，以及 1150 个过表达的脂质物种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，即使在了解代谢物的结构或基因功能缺失后，CRISPR 干扰策略将建立基因之间的因果关系。我们将进一步测试协作杂交中的脂质缺陷菌株。小鼠揭示新发现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。具有重要生物学意义的脂质，决定毒力、宿主界面和结核分枝杆菌存活的关键结果 vivo，支持结核病诊断和治疗的新方法。