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

项目摘要 主要病原菌脂质毒力因子的化学生物学发现 几十年来，对细菌毒性原因的研究一直集中在基因而不是代谢物上。 遗传学方法已经取得了广泛的成功，现代传染病研究依赖于 主要病原体的基因组图谱。由于缺乏全有机化学物质， 生物学工具，细菌脂质还没有系统地测试它们在毒力中的作用，即使脂质 是与人类宿主的主要界面，在那里它们控制营养流并触发宿主免疫 反应我们发明了一种用于脂质谱分析的质谱平台，以检测几乎所有的可电离脂质。 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 伤寒现在提供了明确的证据，为一般的见解，许多，也许是大多数，脂质在世界上的 细菌病原体，是目前未知的命名化合物。基于在确定毒力方面的成功 因子在两个主要病原体的世界性意义，我们将进行化学生物学方法已知 as forward向前lipidomics脂质组学.具体来说，我们将使用整个生物体质谱分析来发现 在毒性细菌中选择性表达的脂质，在现有的脂质目录中是未知的。然后我们 将化学合成与毒性相关的脂质，并将其与其生物合成基因连接以进行删除 在细菌中使用反向遗传方法。使用基因改造的细菌， 脂质，我们将确定毒力脂质在感染过程中的作用。使用天然相同的合成脂质，我们 将确定泡沫巨噬细胞产生的细胞机制并识别免疫受体 介导宿主反应。我们将根据以下内容创建主要革兰氏阴性病原体组的脂质图谱： 病人菌株建立被忽视的化学生物学领域的细菌毒力。这些基本和 翻译研究将支持开发新的前向脂质组学方法来诊断和 重大传染病的治疗。

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.