The role of mycobacterial efflux pumps in virulence and antibiotic tolerance

 描述（由申请人提供）：本提案描述了一项为期5年的研究和培训计划，该计划将使申请人能够实现其成为独立研究者的目标，在结核病（TB）和分枝杆菌发病机制领域开展临床相关基础和转化研究。他将在基础科学研究的基础上，获得微生物学、细菌遗传学和免疫学方面的知识和技能。与此同时，他将进行研究，重点是细菌外排泵在分枝杆菌毒力和抗生素耐受性中的作用。一个设在西雅图儿童研究所、西雅图生物医学和华盛顿大学的指导小组将指导候选人的培训，该小组在分枝杆菌发病机理、细菌遗传学和传染病方面具有广泛的专业知识。研究计划：尽管有几十年的有效抗生素治疗方案，但结核病在2012年仍导致100多万人死亡。消除结核病的障碍包括治疗需要数月的抗生素，治疗不彻底可能促进耐药结核菌株的发展。最近的研究表明，细菌外排泵有助于结核分枝杆菌（Mtb）的抗生素耐受性和细胞内持久性，因此这两个障碍根除。该项目旨在了解结核分枝杆菌和其他分枝杆菌如何利用外排泵在宿主免疫细胞内生存并产生抗生素耐受性。目标1：确定哪些分枝杆菌外排泵是分枝杆菌细胞内存活、毒力和巨噬细胞诱导的抗生素耐受所必需的。候选人将筛选一个图书馆的存档M。海洋突变体，以确定外排泵是必要的，这些现象。目的2：确定是否需要关键的转录因子的协调调控外排泵在分枝杆菌。基于最近的工作，以揭示结核分枝杆菌的转录调控网络，候选人将确定和测试候选转录因子在调节外排泵介导的耐受性和细胞内生存的作用。目标3：确定外排泵的作用和控制其表达的转录网络在结核分枝杆菌和非结核分枝杆菌（NTM）之间是否保守。申请方将测试Mtb的发现是否适用于NTM，NTM是某些人群中的问题病原体。确定在这些过程中重要的外排泵将促进外排泵抑制剂作为结核病和其他分枝杆菌的新型治疗剂的未来发展。培训部分将有助于确立候选人作为结核病和分枝杆菌发病机制的独立调查员。

 DESCRIPTION (provided by applicant): This proposal describes a 5-year research and training program that will allow the applicant to achieve his goal of becoming an independent investigator conducting clinically relevant basic and translational research in the area of tuberculosis (TB) and mycobacterial pathogenesis. He will build on his background in basic science research, gaining knowledge and skills in microbiology, bacterial genetics and immunology. At the same time, he will undertake research focused on the role that bacterial efflux pumps play in mycobacterial virulence and antibiotic tolerance. A mentoring team based at Seattle Children's Research Institute, Seattle BioMed and the University of Washington, with broad expertise in mycobacterial pathogenesis, bacterial genetics and infectious diseases will guide the candidate's training. Research Plan: TB killed over one million people in 2012 despite decades of available, effective antibiotic regimens. Barriers to eradication of TB include that treatment requires many months of antibiotics and that incomplete treatment may promote development of drug resistant TB strains. Recent studies demonstrate that bacterial efflux pumps contribute to antibiotic tolerance and intracellular persistence of Mycobacterium tuberculosis (Mtb) and therefore to both these barriers to eradication. This project seeks to understand how Mtb and other mycobacteria employ efflux pumps to survive within host immune cells and develop antibiotic tolerance. Aim 1: Determine which mycobacterial efflux pumps are required for intracellular survival, virulence and macrophage induced antibiotic tolerance of mycobacteria. The candidate will screen a library of archived M. marinum mutants to identify efflux pumps are necessary for these phenomena. Aim 2: Determine if key transcription factors are required for the coordinated regulation of efflux pumps in mycobacteria. Based on recent work to uncover the transcriptional regulatory network of Mtb, the candidate will identify and test the role of candidate transcription factors in regulating efflux pump mediated tolerance and intracellular survival. Aim 3: Determine if the roles of efflux pumps and the transcriptional networks that control their expression are conserved between Mtb and nontuberculosis mycobacteria (NTM). The applicant will test whether findings in Mtb can be applied to NTM, which are problematic pathogens in certain populations. Identifying efflux pumps important in these processes will facilitate future development of efflux pump inhibitors as novel therapeutic agents for TB and other mycobacteria. The training component will help to establish the candidate as an independent investigator in TB and mycobacterial pathogenesis.