Targeting trans-translation to kill M. tuberculosis non-replicating persister cells

项目摘要/摘要 出现对大多数或所有可用抗生素具有抗药性的结核分枝杆菌(MTB)菌株 给结核病的治疗带来了严重的问题。开发药物的关键障碍之一是 对这些致命的结核分枝杆菌菌株有效的是缺乏新的抗生素靶标。翻译途径 解决停滞的核糖体是药物开发的潜在靶点，因为它是生长所必需的 培养中的结核分枝杆菌。最近发现的小分子翻译抑制物为翻译提供了必要的工具 以确定反式翻译是否可以成为抗生素的靶点。这个项目的长期目标是了解 翻译在结核分枝杆菌中的作用，并利用这一途径开发抗生素。总体目标 这项建议的目的是评估反式翻译在体内感染结核分枝杆菌过程中的作用。的中心假说 这项工作是，在结核杆菌感染过程中抑制反式翻译将导致所有细菌种群的死亡， 包括活跃复制的细胞和非复制的过敏性杆菌。建议的理由是 研究表明，将反式翻译作为抗结核病靶标的确认将导致新的 抗生素将大大缩短结核病的治疗时间，并导致感染组织的消毒。这个 拟议中的研究也将对对结核杆菌的基本科学理解做出重大贡献 通过确定转译在这一有机体中的作用而产生的生理学。这项建议的具体目的是 确定结核分枝杆菌中反式翻译抑制物的分子靶点，以评估反式-翻译抑制物的基本作用 翻译对宿主相关条件下结核分枝杆菌存活的影响，并表征其抗结核活性 酰氨基恶二唑类化合物。拟议的实验使用了基因工具和小分子抑制剂 翻译，以确定在接近正常MTB的条件下是否需要翻译 生理学，如巨噬细胞的生长和小鼠感染期间。这些实验的结果将 为理解细菌中反式翻译在应激和感染过程中的作用以及 确定这一途径是否可以作为药物开发的靶点。生化和遗传方法将是 用于确定活性化合物的分子靶标，结构研究将揭示其分子基础 活跃度。这些研究的结果将使未来对候选药物进行高水平的优化。通过将目标定位为 尚未用于抗生素开发的途径，该项目将产生新的化合物，可以 单独使用或与现有的结核病治疗方法结合使用。

PROJECT SUMMARY/ABSTRACT The emergence of Mycobacterium tuberculosis (MTB) strains that are resistant to most or all available antibiotics has created a severe problem for treating tuberculosis. One of the critical impediments to developing drugs that are effective against these deadly MTB strains is the lack of new antibiotic targets. The trans-translation pathway for resolving stalled ribosomes is a potential target for drug development because it is required for growth of MTB in culture. The recent identification of small molecule inhibitors of trans-translation provides essential tools to determine if trans-translation can be an antibiotic target. The long-term goal of this project is to understand the role of trans-translation in MTB and to exploit this pathway for antibiotic development. The overall objective of this proposal is to evaluate the role of trans-translation during MTB infection in vivo. The central hypothesis of this work is that inhibition of trans-translation during MTB infection will lead to death of all populations of bacteria, including actively replicating cells and non-replicating persister bacilli. The rationale that underlies the proposed research is that validation of trans-translation as an anti-TB target will lead to rapid development of new antibiotics that will dramatically shorten TB treatment times and lead to sterilization of infected tissues. The proposed research will also make significant contributions to the fundamental scientific understanding of MTB physiology by determining the role of trans-translation in this organism. The specific aims of this proposal are to identify the molecular targets of trans-translation inhibitors in MTB, to evaluate the essential role of trans- translation in survival of MTB under host-relevant conditions, and to characterize the anti-tubercular activity of acylaminooxadiazole compounds. The proposed experiments use genetic tools and small molecule inhibitors of trans-translation to determine if trans-translation is required under conditions that approximate normal MTB physiology, such as growth in macrophages and during infection in mice. The results of these experiments will provide the basis for understanding the role of trans-translation during stress and infection in bacteria as well as determining if this pathway can be targeted for drug development. Biochemical and genetic approaches will be used to identify the molecular targets of active compounds, and structural studies will reveal the molecular basis of activity. Results from these studies will allow future high-level optimization of drug candidates. By targeting a pathway that has not been used for antibiotic development, this project will yield new compounds that can be used individually or in combination with existing tuberculosis therapies.