Burkholderia ssp. Novel Therapeutic Approaches

导言。假鼻疽伯克霍尔德氏菌(BP)是生物武器和生物恐怖主义的主要候选者。 因为它很容易获得(从流行区的环境中)、传播和准备(健壮) 在简单的介质上快速生长的生物体)，容易散布，具有高传染性(气雾化和 水污染)，并导致一种迅速发展的严重疾病(类鼻疽病)，死亡率高， 即使得到了正确的诊断。由于BP的内在原因，类鼻炎的治疗非常复杂 对大多数类别的抗生素产生抗药性。目前的治疗方法很昂贵，包括10天的静脉注射 给予头孢菌素类药物，然后口服根除疗法，两至四种药物治疗 3-5个月。因此，感染大量人口在经济和社会发展方面将是灾难性的。 人力成本。在这个项目中，我们将表征外排泵的阻力、结瘤和细胞分裂。 (RND)家族，可能是BP的内在和获得性抗生素耐药的主要参与者。使用强大的 分子和基因工具，我们将表征和评估这些泵和 构建每个都表达一个外排泵的菌株，作为外排泵抑制剂(EPI)发现的工具。我们 然后将筛选EPI化合物文库，项目目标是发现EPI先导化合物。 希望这些努力最终将为类鼻炎提供新的治疗方法，通过增加 现有药物疗法的有效性，并使人们能够使用更便宜和容易获得的药物。 项目交互。本项目II.A.4(Schweizer，PI)将与其他RCE有重要的互动 项目和核心设施。在这个项目中开发的遗传工具将导致与其他 涉及Burkholderia SSP的项目。及其突变体，包括II C.1，毛皮调节基因(Vasil，Pi)，II C2， 氧化和亚硝化应激(Voskuil，PI)，II.C.3，依赖O2的宿主防御(Vazquez-Torres，PI)，II.C.4， 铁摄取(Holmes，PI)和II.A.1，先天免疫在肺部感染中的作用(Dow和Bosio，Co-Pis)。 外排突变体和Pump高表达菌株将成为启动的药物发现努力的有力工具 M.A.2项目，针对选定代理的抗感染药物(Crick和Brennan，Co-Pis)。精选代理档案 Core III.D(Robison，Director)将提供细菌菌株和临床分离株；基因组学/蛋白质组学 CORE III.C.(Slayden，董事)将提供HTS筛选设施，以及进入化合物图书馆和 微阵列。

Introduction. Burkholderia pseudomallei (BP) is a prime candidate for use in biowarfare and bioterrorism because it can be easily obtained (from the environment in endemic areas), propagated and prepared (robust organism rapidly growing on simple media), easily dispersed with high infectivity (aerosolization and contamination of water), and causes a rapidly developing, severe disease (melioidosis) with high mortality, even when properly diagnosed. Treatment of melioidosis is greatly complicated due to BP's intrinsic resistance to most classes of antibiotics. Current therapies are expensive and include 10 day intravenous administration of cephalosporins, followed by oral eradication therapy with a regimen of two to four drugs for 3-5 months. Infections of a large number of people would therefore be disastrous in terms of financial and human costs. In this project we will characterize efflux pumps of the resistance nodulation and cell division (RND) family, the suspected main players in BPs intrinsic and acquired antibiotic resistance. Using powerful molecular and genetic tools, we will characterize and assess the clinical significance of these pumps and construct strains that each express a single efflux pump as a tool for efflux pump inhibitor (EPI) discovery. We will then screen compound libraries for EPIs, with the project goal being the discovery of EPI lead compounds. It is hoped that these efforts will ultimately provide new therapeutic approaches for melioidosis by increasing the effectiveness of existing drug therapies, and by enabling the use of cheaper and readily available drugs. Project interactions. This project II.A.4 (Schweizer, PI) will have significant interactions with other RCE projects and core facilities. Genetic tools developed in this project will lead to obvious interactions with other projects involving Burkholderia ssp. and its mutants, including II.C.1, Fur-regulated genes (Vasil, PI), II.C.2, oxidative and nitrosative stress (Voskuil, PI), II.C.3, O2-dependent host defenses (Vazquez-Torres, PI), II.C.4, iron uptake (Holmes, PI) and II.A.1, role of innate immunity in pulmonary infection (Dow and Bosio, Co-Pis). Efflux mutants and pump overexpressing strains will be powerful tools for the drug discovery efforts initiated in project M.A.2., anti-infectives against select agents (Crick and Brennan, Co-Pis). The Select Agents Archive Core III.D (Robison, Director) will provide bacterial strains and clinical isolates; and the Genomics/Proteomics Core III.C. (Slayden, Director) will provide HTS screening facilities, and access to compound libraries and microarrays.