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

研究大肠杆菌中多药物外排泵和多糖胶囊的新型协同调节

• 批准号: 10078238
• 负责人: Mehreen Arshad
• 金额: $ 18.85万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-18 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078238
• 关键词: Address; Ambulatory Care; Animal Model; Anti-Infective Agents; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Award; Bacteria; Binding; Binding Sites; Biogenesis; Bladder; Centers for Disease Control and Prevention (U.S.); Childhood; Communicable Diseases; Complement; DNA; Deuterium; Disease; Down-Regulation; Drug Efflux; Effectiveness; Escherichia coli; Faculty; Foundations; Funding; Gene Expression; Genes; Genetic Transcription; Goals; Green Fluorescent Proteins; Hydrogen; Immune system; Infection; K-Series Research Career Programs; Knock-out; Knowledge; Laboratories; Ligand Binding; Ligands; Mass Spectrum Analysis; Measurement; Medical; Mentorship; Modeling; Molecular; Molecular Conformation; Morbidity - disease rate; Multi-Drug Resistance; Mus; Mutagenesis; Organism; Outpatients; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Phenotype; Polysaccharides; Positioning Attribute; Predisposition; Promoter Regions; Proteins; Public Health; Regulation; Regulatory Pathway; Research; Research Institute; Research Personnel; Research Proposals; Role; Seeds; Sepsis; Site; Specimen; Support System; Testing; Training; Transcription Repressor; Universities; Up-Regulation; Urinary tract infection; Uropathogenic E. coli; Virulence; Virulence Factors; Visit; Work; Xenobiotics; antimicrobial; attenuation; bacterial genetics; capsule; career; chemical genetics; chromatin immunoprecipitation; community-acquired UTI; cost estimate; efflux pump; experimental study; genetic approach; genome-wide; immune clearance; inhibitor/antagonist; innovation; interest; microbial; microbial genomics; mortality; mouse model; mutant; novel; novel therapeutics; pathogen; programs; protein structure; public health emergency; receptor; response; skills; small molecule; small molecule inhibitor; transcriptome

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.

项目概要 在此申请中，我请求职业发展奖来支持我的研究计划，标题为 “研究大肠杆菌中多药物外排泵和多糖胶囊的新型协同调节。”我会 在杜克大学 Patrick Seed 博士的指导下开展这项工作。本次活动的目的 提出这个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目标 针对革兰氏阴性微生物的抗菌剂类别。泌尿道致病性大肠杆菌 (UPEC) 是最常见的泌尿道致病性大肠杆菌之一 常见的革兰氏阴性菌，也是尿路感染 (UTI) 的主要原因。然而，抗生素的快速增长 耐药性威胁着所有常用抗生素的有效性。此外，多国的全球崛起 耐药 (MDR) UPEC 克隆亚型 ST131 已被 疾病控制中心。这共同强调了识别新抗菌药物的迫切需要 策略。由于大多数社区获得性尿路感染病例发生在免疫系统完整的患者中， 治疗此类感染的替代方法是使细菌对自然免疫敏感 清除。我们已经鉴定出一种高效的大肠杆菌胶囊生物合成小分子抑制剂，其目标是 MDR 外排泵的转录调节因子。该提案的中心假设是该胶囊 抑制剂独特地与外排泵转录调节因子结合以控制大肠杆菌多糖胶囊 通过一种新的共同调节途径表达，导致毒力降低并增加 对宿主免疫系统的易感性。本研究将通过调查结合来检验这一假设 通过识别共调节途径的组成部分，将胶囊抑制剂抑制到转录调节因子 并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 将提供分子微生物发病机制、微生物基因组学领域的指导和专业知识 和尿路感染的小鼠模型。我自 2011 年起作为儿科传染病研究员与 Seed 博士一起工作 并将作为初级教师继续这样做。杜克大学是一所世界一流的研究机构，拥有众多 初级调查员的支持系统对于我向独立的过渡至关重要 研究者。我的长期职业目标是独立资助领导我自己的实验室，专注于使用 分子方法来确定细菌发病机制和抗菌药物耐药性的机制 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。该奖项将 使我能够在迄今为止所建立的研究基础上再接再厉，并做好准备 向独立过渡。

项目成果

ESBL Escherichia coli Isolates Have Enhanced Gut Colonization Capacity Compared to Non-ESBL Strains in Neonatal Mice.

• DOI: 10.1128/spectrum.00582-22
• 发表时间: 2022-10-26
• 期刊: Microbiology spectrum
• 影响因子: 3.7

Mechanisms and modulation of sepsis-induced immune dysfunction in children.

• DOI: 10.1038/s41390-021-01879-8
• 发表时间: 2022-01
• 期刊: Pediatric research
• 影响因子: 3.6
• 作者: Mithal LB;Arshad M;Swigart LR;Khanolkar A;Ahmed A;Coates BM
• 通讯作者: Coates BM

The Role of Probiotics, Prebiotics and Synbiotics in Combating Multidrug-Resistant Organisms.

• DOI: 10.1016/j.clinthera.2020.06.011
• 发表时间: 2020-09
• 期刊: Clinical therapeutics
• 影响因子: 3.2
• 作者: Newman AM;Arshad M
• 通讯作者: Arshad M

The Gut of Healthy Infants in the Community as a Reservoir of ESBL and Carbapenemase-Producing Bacteria.

社区中健康婴儿的肠道是 ESBL 和产碳青霉烯酶细菌的储存库。

• DOI: 10.3390/antibiotics9060286
• 发表时间: 2020
• 期刊: Antibiotics (Basel, Switzerland)
• 作者: Saleem,AliF;Allana,Ahreen;Hale,Lauren;Diaz,Alondra;Salinas,Raul;Salinas,Cristina;Qureshi,ShahidaM;Hotwani,Aneeta;Rahman,Najeeb;Khan,Asia;Zaidi,AnitaK;Seed,PatrickC;Arshad,Mehreen
• 通讯作者: Arshad,Mehreen