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.

结核分枝杆菌是一种很难杀死的细菌。结核分枝杆菌感染的灭菌 活动性结核病患者需要至少 6 个月的治疗，并服用 3-4 种药物 日常的。这种顽强的生命力是任何其他人类细菌病原体都无法比拟的。结核分枝杆菌有 进化出面对药物和宿主免疫反应时生存的策略。目前的 该领域的教条表明，生存是细菌进入药物的能力的结果 耐受状态，它是非复制和休眠的，但显然是可行的。如果及早停药 或者免疫系统受到损害，休眠的细菌可能会复活并继续发展为活动性疾病。 作为该计划项目的一部分，我们正在投入时间和资源来寻找“阿喀琉斯之踵” 使用分枝杆菌遗传学研究休眠细菌。对于这个提案，我们产生了很高的评价 专门转导的吞吐量方法，可用于破坏任何非必要的 基因。此外，我们还使用条件表达和专门的转导 开发 CESTET，一种证明必要性并研究分枝杆菌细胞命运的方法 基因产物的耗尽。已开发出体外和体内测定法来筛选 必需基因和持久性因素。该集团拥有良好的成功记录 解决结构已经解决了 PDB 中 70% 的 Mtb 结构，我们将以此为基础 成功解决新的基本或持久性目标的结构。使用结构 信息，将进行虚拟配体筛选以识别新的抑制剂。因此，本次活动的目标 项目旨在验证目标，解析其三维结构，并提出新型抑制剂 进一步开发药物以从根本上缩短治疗结核病的时间。