Anaerobic shock as a novel treatment for tuberculosis

描述(由申请人提供) 摘要：结核病每年导致200万人死亡。多重耐药(MDR)感染的日益流行和广泛/完全耐药(XDR/CDR)结核杆菌的出现使目前的化疗效果越来越差。这项建议的长期目标是开发一种新的结核病治疗方法，基于我们最近观察到的厌氧性休克导致结核分枝杆菌快速和广泛的细胞裂解(例如，细菌存活率在5分钟内下降)。我们将探索两个大体方向。一种是用厌氧气体治疗受感染宿主的肺部。首先，感染结核分枝杆菌的兔，包括MDR和XDR临床分离株，将被用于测试体外优化的厌氧混合气体的有效性和安全性。将寻求额外资金的临床试验将基于我们的兔子工作进行设计。我们这项工作的总体成果应该是提供一种医疗程序，在几小时甚至几分钟内实现痰杆菌阳性转化为阴性，而不是传统联合化疗所需的几周时间。第二个方向集中在厌氧休克介导的快速细胞裂解的分子机制上。基因、基因组和生化方法将被用来利用直接的浊度和荧光分析来识别与厌氧休克诱导的细胞裂解有关的基因和编码蛋白。还将构建和筛选体内环肽文库，以寻找与厌氧性休克类似的自溶诱导活性。从这项工作的第二部分获得的信息将用于设计用于分枝杆菌细胞裂解的小分子激活剂的高通量筛选。该项目的成功将给结核病治疗带来革命性的变化，并产生一种新药，无论其耐药性(MDR或XDR)或生理状态(生长或休眠)如何，都能迅速治愈结核病。 公共卫生相关性：结核病感染了世界三分之一的人口，每年导致200万人死亡。日益严重的耐药性问题可能很快就会使目前所有可用的治疗方案失效。本计划旨在开发一种新的、快速和高效的治疗结核病的方法，通过诱导结核分枝杆菌的自毁自溶来实现。

DESCRIPTION (Provided by the applicant) Abstract: Tuberculosis kills 2 million people every year. The increasing prevalence of multiple drug- resistant (MDR) infections and the emergence of extensively/completely drug-resistant (XDR/CDR) tubercle bacilli are making currently chemotherapies less and less effective. The long-term goal of this proposal is to develop a novel therapy for tuberculosis based on our recent observation that anaerobic shock causes rapid and extensive cell lysis of Mycobacterium tuberculosis (e.g. > 105 drop in bacterial survival within 5 min). Two general directions will be explored. One is treatment of lungs of infected hosts with anaerobic gas. Initially, rabbits infected with M. tuberculosis, including MDR and XDR clinical isolates, will be used to test the efficacy and safety of anaerobic gas mixtures optimized in vitro. Clinical trials, for which additional funding will be sought, will be designed based on our rabbit work. Our overall achievement of this line of work should provide a medical procedure that will achieve a positive to negative sputum bacilli conversion in hours, or perhaps even minutes, rather than the weeks required for traditional combination chemotherapy. The second direction focuses on the molecular mechanisms underlying anaerobic shock-mediated rapid cell lysis. Genetic, genomic, and biochemical approaches will be used to identify relevant genes and the encoded proteins involved in anaerobic shock-induced cell lysis using straight-forward turbidity and fluorescence assays. An in vivo cyclic peptide library will also be constructed and screened for autolysis- inducing activities similar to that conferred by anaerobic shock. Information gained from this second part of the work will be used to design high throughput screens for small-molecule activators of mycobacterial cell lysis. The success of this project will revolutionize tuberculosis therapy and generate a novel drug that would rapidly cure tuberculosis regardless of its drug- resistance profile (MDR or XDR) or physiological status (growing or dormant). Public Health Relevance: Tuberculosis infects a third of the world's population and kills 2 million people a year. The increasing problem of drug resistance may soon make all currently available treatment options ineffective. The present program seeks to develop a novel, rapid, and highly efficient treatment of tuberculosis upon induction of self-destructive autolysis of Mycobacterium tuberculosis.