Genetic approaches to protein-protein interactions mediating antibiotic resistanc

描述（由申请人提供） 摘要：抗生素耐药菌，如万古霉素耐药肠球菌（VRE）和耐甲氧西林金黄色葡萄球菌（MRSA），是医院获得性感染的主要原因，是不断升级的健康危机的驱动力。我们将通过无偏见的遗传选择利用细菌遗传学的力量来帮助解决新兴的抗生素耐药性问题：1）全面鉴定导致抗生素耐药性的细胞途径中的蛋白质-蛋白质相互作用; 2）发现使这些蛋白质-蛋白质相互作用失效的小分子。拟议的研究将共同利用我们在遗传策略开发方面的专业知识，以及我们对阐明细菌抗生素耐药性基本机制的持续兴趣。通过关注蛋白质-蛋白质相互作用，这项研究有望：1）揭示抗生素耐药机制的潜在生物学及其整合到细菌宿主的生理过程中的新见解; 2）定义新的靶标（以蛋白质-蛋白质相互作用的形式）用于治疗由耐药病原体引起的感染的创新疗法;和3）鉴定具有独特作用模式的新型小分子候选药物。我们的实验设计具有关键的战略优势。例如，我们的分析将在耐药细菌宿主的天然背景下进行（例如，而不是通过对分离的蛋白质进行体外筛选），这将使我们能够捕获动态细胞过程或翻译后修饰对关键蛋白质-蛋白质相互作用的任何潜在的、但尚不为人知的影响。此外，我们将采用强大的遗传选择，能够快速筛选巨大的文库，以揭示即使是罕见的命中，根据定义，在生理环境中是功能性的。总的来说，这些策略将能够发现未知的，不可预测的和新颖的生物学见解，这些见解是传统手段无法获得的，将被用来发现对耐药细菌感染有效的新候选疗法。 公共卫生相关性：抗生素耐药细菌，如万古霉素耐药肠球菌（VRE）和耐甲氧西林金黄色葡萄球菌（MRSA），是医院获得性感染的主要原因，是不断升级的健康危机的驱动力。这项研究承诺：1）揭示抗生素耐药性机制的潜在生物学的新见解，这将促进耐药病原体引起的感染的新治疗方法的开发; 2）为这些创新疗法定义新靶点; 3）确定具有独特作用模式的新型小分子候选药物。

DESCRIPTION (Provided by the applicant) Abstract: Antibiotic-resistant bacteria, such as vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA), are major causes of hospital-acquired infections and are driving forces of an escalating health crisis. We will help address the burgeoning antibiotic resistance problem by leveraging the power of bacterial genetics via unbiased genetic selections to: 1) comprehensively identify protein-protein interactions in cellular pathways that result in antibiotic resistance; and 2) discover small molecules that disable these protein-protein interactions. The proposed research will jointly exploit our expertise in the development of genetic strategies and our ongoing interest in elucidating the fundamental mechanisms of bacterial antibiotic resistance. By focusing on protein-protein interactions, this research promises to: 1) reveal new insights into the underlying biology of antibiotic resistance mechanisms and their integration into the physiological processes of the bacterial host; 2) define new targets (in the form of protein-protein interactions) for innovative therapeutics to treat infections caused by drug-resistant pathogens; and 3) identify novel small-molecule drug candidates with unique modes of action. Our experimental design possesses critical strategic advantages. For example, our analyses will be done within the native context of the drug-resistant bacterial host (e.g., not by in vitro screens on isolated proteins), which will enable us to capture any potential, but as yet unknown, effects of dynamic cellular processes or post-translational modifications on key protein-protein interactions. Furthermore, we will employ powerful genetic selections capable of rapidly sifting through immense libraries to reveal even rare hits that, by definition, are functional in a physiological context. Collectively, these strategies will enable the discovery of unknown, unpredictable, and novel biological insights, not accessible by conventional means, that will be exploited to discover new candidate therapeutics with efficacy against drug-resistant bacterial infections. Public Health Relevance: Antibiotic-resistant bacteria, such as vancomycin-resistant enterococci (VRE) and methicillinresistant Staphylococcus aureus (MRSA), are major causes of hospital-acquired infections and are driving forces of an escalating health crisis. This research promises to: 1) reveal new insights into the underlying biology of antibiotic resistance mechanisms that will facilitate the development of new treatments for infections caused by drug-resistant pathogens; 2) define new targets for these innovative therapeutics; and 3) identify novel small-molecule drug candidates with unique modes of action.