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

项目总结/摘要 结核分枝杆菌（MTB）菌株的出现对大多数或所有可用的抗生素都具有耐药性 给肺结核的治疗带来了严重的问题。开发药物的一个关键障碍是， 有效对抗这些致命的MTB菌株的关键是缺乏新的抗生素靶点。反式翻译途径 解决停滞的核糖体是药物开发的潜在目标，因为它是生长所必需的。 文化中的MTB。最近发现的小分子反式翻译抑制剂提供了必要的工具 以确定反式翻译是否可以作为抗生素的靶点。这个项目的长期目标是了解 反式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 本研究的主要目的是评价MTB体内感染过程中反式翻译的作用。的中心假设 这项工作是在MTB感染期间抑制反式翻译将导致所有细菌群体的死亡， 包括活跃复制的细胞和非复制的持留杆菌。建议的基本原理 研究表明，反式翻译作为抗结核病靶点的验证将导致新的 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。的 拟议中的研究也将对MTB的基本科学认识做出重大贡献 通过确定这种生物体中反式翻译的作用来研究生理学。这项建议的具体目标是 确定MTB中反式翻译抑制剂的分子靶点，以评估反式- 翻译在宿主相关条件下MTB存活中的作用，并表征 酰基氨基恶二唑化合物。拟议的实验使用遗传工具和小分子抑制剂， 反式翻译，以确定在接近正常MTB的条件下是否需要反式翻译 生理学，如巨噬细胞中的生长和小鼠感染期间。这些实验的结果将 为理解细菌在应激和感染过程中反式翻译的作用提供了基础， 以确定该途径是否可以用于药物开发。生物化学和遗传学方法将是 用于确定活性化合物的分子靶点，结构研究将揭示分子基础 的活动。这些研究的结果将使未来的候选药物的高水平优化。通过靶向 该途径尚未用于抗生素开发，该项目将产生新的化合物， 单独使用或与现有的结核病疗法联合使用。

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.