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

项目摘要 主要致病菌中脂类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上，而不是代谢物上。 基因方法在很大程度上是成功的，现代传染病研究依靠 基本上取决于主要病原体的基因组图谱。由于缺乏全生物体的化学物质 生物学工具，细菌脂类还没有系统地测试它们在毒力中的作用，即使脂类 是与人类宿主的主要接口，在那里它们控制营养流动并触发宿主免疫 回应。我们发明了一种用于脂类分析的质谱学平台，可以检测几乎所有可电离的脂类 细菌细胞在2小时内。结核分枝杆菌和肠道沙门氏菌血清型的实验研究 伤寒现在为普遍的洞察力提供了明确的证据，即世界上许多，也许是大多数的脂质 细菌病原体，目前作为命名化合物尚不为人所知。基于在鉴定毒力方面的成功 在两种具有世界意义的主要病原体因素中，我们将开展一种已知的化学生物学方法 这是一种前沿性的脂质组学。具体地说，我们将使用整个生物体的质谱图来发现 在致病细菌中选择性表达的脂类，在现有的脂类目录中未知。然后，我们 将化学合成与毒力相关的脂类，并将它们与其生物合成基因联系起来进行删除 在使用反向遗传方法的细菌中。使用定义有缺陷的转基因细菌 脂类，我们将确定毒力类脂在感染过程中的作用。使用天然相同的合成脂类，我们 将确定泡沫巨噬细胞产生的细胞机制并识别免疫受体 这是宿主反应的中介。我们将根据以下内容创建主要革兰氏阴性病原菌群的脂谱 患者菌株构建了被忽视的细菌毒力化学生物学领域。这些基本的和 翻译研究将支持开发新的正向脂组学方法来诊断和 重大传染病的救治。

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.