Chemical Biological Discovery of Lipid Virulence Factors in the Major Bacterial Pathogens

主要细菌病原体中脂质毒力因子的化学生物学发现

基本信息

批准号：
10518252
负责人：
DAVID Branch MOODY
金额：
$ 64.25万
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：
2022
资助国家：
美国
起止时间：
2022-06-22 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10518252
关键词：
Address Autophagocytosis Bacteria Bacterial Chromosomes Biological Biological Process Biology Catalogs Cell model Cells Cellular Assay Chemical Structure Chemicals Chromosome Mapping Communicable Diseases Cord Factors Data Databases Detection Development Diagnosis Disease Enteral Environment Evolution Feedback Foamy Macrophage Functional disorder Gene Deletion Generations Genes Genomics Genus Mycobacterium Gram-Negative Bacteria Hour Human Immune response Immunologic Receptors Infection Infectious Diseases Research Laboratories Link Lipids Literature Location Lysosomes Maps Mass Spectrum Analysis Measures Mediating Membrane Methods Modernization Molecular Mus Mycobacterium tuberculosis Names Nature Nucleosides Nutrient Organism Patients Phenotype Phospholipids Process Production Proteins Proteomics Role Salmonella Salmonella enterica Salmonella typhi Science Signal Transduction Study Section System T cell response Testing Transcript Virulence Virulence Factors Virulent Whole Organism base comparative comparative genomics experience experimental study forward genetics gene discovery gene function genetic approach in vivo inhibitor insight lipidome lipidomics macrophage metabolomics mycobacterial novel strategies overexpression pathogen pathogenic bacteria prenyl receptor response reverse genetics selective expression success tool transcriptomics translational study

项目摘要

Project Summary Chemical Biological Discovery of Lipid Virulence Factors in the Major Bacterial Pathogens For decades, the search for the causes of bacterial virulence has focused on genes rather than metabolites. Genetic approaches have been broadly successful, and modern infectious disease research relies fundamentally on genomic maps of the major pathogens. Owing to the lack of whole-organism chemical biology tools, bacterial lipids have not been systematically tested for their roles in virulence, even though lipids are the primary interface with the human host, where they control nutrient flow and trigger host immune response. We invented a mass spectrometry platform for lipid profiling to detect nearly all ionizable lipids in a bacterial cell within 2 hours. Experiments on Mycobacterium tuberculosis and Salmonella enterica serovar Typhi now provide clear evidence for the general insight that many, perhaps the majority, of lipids in the world's bacterial pathogens, are currently unknown as named compounds. Based on successes in identifying virulence factors in two major pathogens of worldwide significance, we will carry out a chemical biology approach known as forward lipidomics. Specifically, we will use whole organism mass spectrometry profiling to discover the lipids that are selectively expressed in virulent bacteria and are unknown in existing lipid catalogs. Then, we will chemically synthesize the virulence associated lipids and link them to their biosynthetic genes for deletion in bacteria using reverse genetic approaches. Using genetically modified bacteria that are deficient in defined lipids, we will determine the roles of virulence lipids during infection. Using nature identical synthetic lipids, we will determine the cellular mechanisms of generation of foamy macrophages and identify immune receptors that mediate host response. We will create lipid maps of the major Gram negative pathogen groups based on patient strains to build the overlooked field of chemical biology of bacterial virulence. These basic and translational studies will support the development new forward lipidomics approaches to the diagnosis and treatment of major infectious diseases.

项目摘要主要细菌病原体中脂质毒力因子的化学生物学发现几十年来，寻找细菌毒力的原因一直集中在基因而不是代谢物上。遗传方法已广泛成功，现代传染病研究依赖从根本上讲，主要病原体的基因组图。由于缺乏全生物化学生物学工具，细菌脂质尚未系统地测试其在毒力中的作用，即使脂质是人类宿主的主要界面，它们控制营养流并触发宿主免疫回复。我们发明了一个质谱平台，用于脂质分析，以检测几乎所有可离子脂质细菌细胞在2小时内。结核分枝杆菌和沙门氏菌肠血清的实验 Typhi现在为一般见解提供了明确的证据，即世界上许多脂质的脂质细菌病原体目前未知为命名化合物。基于确定毒力的成功全球意义的两种主要病原体的因素，我们将采取一种化学生物学方法已知作为前向脂肪组学。具体而言，我们将使用整个生物体质谱分析来发现在毒细菌中选择性表达且在现有脂质目录中未知的脂质。然后，我们将通过化学合成毒力相关的脂质，并将其链接到其生物合成基因以进行缺失在细菌中使用反向遗传方法。使用定义不足的转基因细菌脂质，我们将确定感染过程中毒力脂质的作用。使用自然相同的合成脂质，我们将确定泡沫巨噬细胞产生的细胞机制并鉴定免疫受体介导宿主反应。我们将基于患者菌株建立细菌毒力的化学生物学领域。这些基本和翻译研究将支持开发新的前向脂肪组学的方法来诊断和治疗主要的传染病。