Structural Genomics of Persistence Targets from M. tuberculosis

结核分枝杆菌持久性靶点的结构基因组学

• 批准号: 7063006
• 负责人: JAMES C SACCHETTINI
• 金额: $ 77.86万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7063006
• 关键词: Mycobacterium tuberculosis; adenosine triphosphate; adenosinetriphosphatase; bacterial genetics; biosynthesis; cell differentiation; chemical structure function; clinical research; drug administration rate /duration; drug discovery /isolation; drug resistance; enzyme mechanism; fatty acid biosynthesis; gene deletion mutation; gene induction /repression; high throughput technology; hydroxy fatty acid; laboratory mouse; latent bacterial disease; posttranscriptional RNA processing; protein transport; tissue /cell culture; transfection /expression vector

Mycobacterium tuberculosis is a tough bug to kill. The sterilization of a M. tuberculosis infection in a patient with active tuberculosis requires a minimum of 6 months of treatment with 3-4 drugs taken daily. This tenacity is unparalleled in any other human bacterial pathogen. M. tuberculosis has evolved strategies to survive in the face of drugs and the host immune responses. The current dogma in the field suggests that survival is the result of the bacteria's ability to enter into a drug tolerant state, where it is non-replicative and dormant, but clearly viable. If drugs are stopped early or the immune system is compromised, dormant bacteria can revive and go on to active disease. As part of the program project, we are dedicating our time and resources to find the "Achilles' heel" of the dormant bacteria using mycobacterial genetics. For this proposal, we have generated a high throughput methodology of specialized transduction, which can be used to disrupt any nonessential gene. In addition, we have used conditional expression and specialized transduction to develop CESTET, a method to prove essentiality and to study a mycobacterial cell's fate upon depletion of the gene product. In vitro and in vivo assays have been developed to screen for essential genes and persistence factors. The group has a proven track record of successfully solving structures having solved 70% of the Mtb structures in the PDB, and we will build on this success to solve the structures of the new essential or persistence targets. Using the structural information, virtual ligand screens will be performed to identify new inhibitors. Thus, the goal of this Project is to validate targets, solve their three dimensional structures, and propose novel inhibitors for further drug development to radically shorten the time to treat TB.

结核分枝杆菌是一个很难杀死的虫子。结核分枝杆菌感染的灭菌 活性结核病的患者至少需要接受3-4种药物治疗6个月 日常的。这种韧性在任何其他人类细菌病原体中都是无与伦比的。结核分枝杆菌具有 面对药物和宿主免疫反应，进化的策略是生存的。电流 该领域的教条表明生存是细菌进入药物的能力的结果 宽容状态，它是无复制和休眠状态的，但显然可行。如果毒品早点停止 或免疫系统受到损害，休眠细菌可以恢复并继续进行活性疾病。 作为计划项目的一部分，我们正在花时间和资源来找到“阿喀琉斯'脚跟” 使用分枝杆菌遗传学的休眠细菌。对于此提案，我们产生了很高的 专业转导的吞吐量方法，可用于破坏任何不必要 基因。此外，我们已经使用了条件表达和专门的转导 开发CESTET，一种证明和研究分枝杆菌细胞的命运的方法 基因产物的耗竭。已经开发了体外和体内测定以筛选 基本基因和持久性因素。该小组有成功的记录 解决了已解决PDB中70％MTB结构的结构，我们将在此基础上构建 成功解决新基本或持久目标的结构的成功。使用结构 信息，虚拟配体屏幕将进行识别新抑制剂。因此，目标的目标 项目将验证目标，解决其三维结构，并提出新型抑制剂 为了进一步的药物开发，可以从根本上缩短治疗结核病的时间。