Host factors involved in bacterial pathogenesis

参与细菌发病机制的宿主因素

• 批准号: 7548603
• 负责人: NORBERT PERRIMON
• 金额: $ 32.29万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7548603
• 关键词: Affect; Antibiotic Resistance; Appearance; Applications Grants; Bacteria; Cells; DNA Sequence Rearrangement; Data; Development; Dissection; Doctor of Philosophy; Drosophila genus; Environment; Genetic; Genome; Genus Mycobacterium; Growth; Health; Human; Infection; Integration Host Factors; Listeria monocytogenes; Mammalian Cell; Mediating; Membrane; Methodology; Molecular; Mycobacterium Infections; Mycobacterium tuberculosis; Nutrient; Pathogenesis; Phagocytes; Phagocytosis; Play; Population; Process; Property; RNA Interference; Receptor Signaling; Research Personnel; Resistance; Role; SR-B proteins; Secondary to; Signal Transduction; System; Validation; Virulent; antimicrobial; cell killing; functional gain; high throughput screening; in vivo; insight; interest; killings; macrophage; mycobacterial; novel; pathogen; prevent; programs; receptor; research study; trafficking; uptake

The turn of the twentieth century has witnessed the continued appearance of new pathogens as well as the development of antibiotic resistance and re-emergence of old pathogens. Despite the enormous burden that bacterial pathogens impose on human health, there is relatively little understanding of host-pathogen interactions. The hypothesis underpinning this grant proposal is that by studying the mechanisms by which bacteria enter Drosophila cells, replicate and ultimately kill the cells, we will be able to identify evolutionary conserved host factors that are involved in the interaction between pathogens and their hosts. Studies have clearly established that Drosophila S2 cells, which have the properties of macrophages, are an excellent system to study host-pathogen interactions. We will focus our studies on Mycobacterium spp. It is estimated that one-third of the world's population is infected with Mycobacterium tuberculosis, and it kills more people yearly than any other bacterial pathogen. It, like other pathogenic mycobacteria, can convert the normally hostile environment of a macrophage into a niche in which it can effectively replicate. We propose high-throughput full genome screens in Drosophila cells, using the RNA interference (RNAi) methodology, to identify host factors required for entry into and survival within macrophages. Preliminarydata indicatethat M.fortuitum and M. marinum effectively enter and replicate in Drosophila cells, and that we can identify host factors required for these processes. We hypothesize that by elucidating the host factors required for mycobacterial entry and survival within host cells, we will gain an understandingof the common machinery used by intracellular pathogens as well as specific factors that are important for the intracellular strategy employed by mycobacteria. The host factors identified in Drosophila cells will then be used to dissect the mechanisms of bacterial uptake and intracellular survival followed by validation in mammalian systems. Thus, exploiting the powerful genetic strategy of RNAi in Drosophila, we hope to elucidate novel bacterial strategies for intracellular survival that may ultimately allow us to gain functional and mechanistic insight into the interaction between M. tuberculosis and its human host.

二十世纪之交目睹了新病原体的持续出现以及 抗生素耐药性的发展和旧病原体的重新出现。尽管负担很大 细菌病原体对人类健康施加，对宿主病原体的了解相对较少 互动。该赠款提案的基础的假设是，通过研究 细菌进入果蝇细胞，重复并最终杀死细胞，我们将能够识别 进化保守的宿主因子与病原体与其宿主之间的相互作用有关。 研究清楚地表明，具有巨噬细胞特性的果蝇S2细胞是 研究宿主病原体相互作用的出色系统。我们将把研究重点放在分枝杆菌属上。这是 据估计，世界人口中有三分​​之一感染了结核分枝杆菌，它杀死了 每年的人比任何其他细菌病原体都多。它像其他致病性分枝杆菌一样可以转换 通常，巨噬细胞的敌对环境可以有效地复制。我们建议 使用RNA干扰（RNAI）方法论，果蝇细胞中的高通量全基因组筛选， 确定进入巨噬细胞内和生存所需的宿主因素。初步数据指示 M.Forthuitum和M. Marinum有效地进入并复制在果蝇细胞中，我们可以识别宿主 这些过程所需的因素。我们假设通过阐明宿主因素 宿主细胞内的分枝杆菌进入和生存，我们将了解通用机械的一种 由细胞内病原体以及对细胞内策略很重要的特定因素使用 由分枝杆菌雇用。然后，将使用果蝇细胞中确定的宿主因子进行剖析 细菌摄取和细胞内存活的机制，然后在哺乳动物系统中进行验证。 因此，利用果蝇中RNAi强大的遗传策略，我们希望阐明新型细菌 细胞内生存的策略，最终可能使我们获得功能和机械洞察力 结核分枝杆菌与其人类宿主之间的相互作用。