Investigating a Novel Co-regulation of Multi-Drug Efflux Pumps and Polysaccharide Capsule in E. coli

项目总结 在这份申请中，我申请职业发展奖，以支持我的研究计划，标题为 研究一种新型的多重药物外排泵和多糖胶囊在大肠杆菌中的共同调节。这就做 在杜克大学帕特里克·塞德博士的指导下进行这项工作。这样做的目的是 这项提议是出于我对确定毒力特定因素的兴趣，这些因素可以用作新的 针对革兰氏阴性菌的抗菌药类别。尿路致病性大肠埃希菌(UPEC)是最常见的 常见的革兰氏阴性杆菌，也是尿路感染(UTI)的主要原因。然而，迅速崛起的抗生素 耐药性威胁着所有常用抗生素的有效性。此外，多国集团在全球的崛起- 耐药(MDR)UPEC克隆型ST131被世界卫生组织称为对公众健康的紧急威胁 疾病控制中心。总而言之，这突显了确定新的抗菌剂的迫切需要 战略。由于大多数社区获得性尿路感染发生在免疫系统完整的患者和 治疗这种感染的另一种方法是使细菌对自然免疫敏感。 通行证。我们已经确定了一种针对大肠杆菌胶囊生物发生的高效小分子抑制物 MDR外排泵的转录调节因子。这项提议的核心是假设这个胶囊 抑制物与外排泵转录调控因子结合控制大肠杆菌多糖胶囊 通过一种新的共调节途径表达，导致毒力降低和毒力增加 对宿主免疫系统的易感性。这项研究将通过调查两个基因的结合来检验这一假设 通过鉴定共调控途径的组成部分，胶囊抑制剂对转录调节因子的作用 并通过确定转录调节因子在尿路感染的建立和持续中的作用。种子博士 将在分子微生物发病机制、微生物基因组学领域提供指导和专业知识 尿路感染的小鼠模型。自2011年以来，我一直作为儿科传染病研究员与Seed医生共事 并将继续以初级教员的身份这样做。杜克大学是一所世界级的研究机构，拥有众多 初级调查人员的支持系统，这将是我向独立调查人员过渡的关键 调查员。我的长期职业目标是独立出资，领导我自己的实验室，专注于使用 鉴定细菌致病机制和抗菌药耐药性的分子方法 利用这些知识来确定病原体/疾病特异性抗感染药物的新靶点。这一奖项将 使我能够在我到目前为止开发的研究基础上再接再厉，并特别准备好 向独立过渡。

PROJECT SUMMARY In this application, I am requesting a career development award to support my research proposal titled “Investigating a Novel Co-regulation of Multi-Drug Efflux Pumps and Polysaccharide Capsule in E. coli.” I will be undertaking this work under the mentorship of Dr. Patrick Seed at Duke University. The aims of this proposal arose out of my interest in identifying virulence specific factors that can be used as targets for new classes of antimicrobials against Gram negative organisms. Uropathogenic E. coli (UPEC) is one of the most common Gram negative and a major cause of urinary tract infection (UTI). However, rapidly rising antibiotic resistance threatens the effectiveness of all commonly used antibiotics. In addition the global rise of the multi- drug resistant (MDR) UPEC clonal subtype ST131 has been termed an urgent threat to public health by the Centers for Disease Control. Together this underscores the critical need to identify new antimicrobial strategies. Since most cases of community acquired UTI occur in patients with an intact immune system an alternative approach to treating such infections would be to render the bacteria susceptible to natural immune clearance. We have identified a highly potent small molecule inhibitor of E. coli capsule biogenesis that targets a transcriptional regulator of an MDR efflux pump. This proposal centers on the hypothesis that this capsule inhibitor uniquely binds to the efflux pump transcriptional regulator to control E. coli polysaccharide capsule expression through a novel co-regulatory pathway that results in decreased virulence and increased susceptibility to the host immune system. This study will test the hypothesis by investigating the binding of capsule inhibitor to the transcriptional regulator, by identifying the components of the co-regulatory pathway and by determining the role of the transcriptional regulator in establishment and persistence of UTI. Dr. Seed will be providing mentorship and expertise in the field of molecular microbial pathogenesis, microbial genomics and the murine model of UTI. I have worked with Dr. Seed since 2011 as a pediatric infectious disease fellow and will continue to do so as junior faculty. Duke University is a world-class research institute with numerous support systems for junior investigators which will be vital for my transition to being an independent investigator. My long term career goal is to be independently funded leading my own lab focused on using molecular approaches to identify mechanisms of bacterial pathogenesis and antimicrobial resistance and to use this knowledge to identify novel targets for pathogen/disease-specific anti-infectives. This award will enable me to build upon the research foundation I have developed thus far and be exceptionally well poised to transition to independence.