Antibiotic tolerance by multidrug resistant uropathogenic Escherichia coli during quiescence

多重耐药尿路致病性大肠杆菌在静止期间的抗生素耐受性

• 批准号: 10381489
• 负责人: Jodi Lynn Camberg
• 金额: $ 19.69万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10381489
• 关键词: Agar; Animals; Antibiotic Resistance; Antibiotic Therapy; Antibiotic susceptibility; Antibiotics; Bacteria; Biological Assay; Biological Models; Bladder; Bladder Urothelial Cell; Blood; Cell Culture Techniques; Cell physiology; Cells; Clinical; Collaborations; Epithelial Cells; Escherichia coli; Escherichia coli Infections; Foundations; Frequencies; Genetic; Glucose; Goals; Growth; Human; In Vitro; Infection; Lead; Measures; Modeling; Multi-Drug Resistance; Multidrug Resistance Gene; Organism; Outcomes Research; Patients; Peptidoglycan; Pharmaceutical Preparations; Phenotype; Phylogenetic Analysis; Physiological; Physiology; Population; Predisposition; Prevalence; Proliferating; Recurrence; Reporting; Research; Resistance; Signal Transduction; Testing; Time; Translating; Urinary tract infection; Urine; Uropathogenic E. coli; Vesicle; Virulent; Woman; antibiotic tolerance; bactericide; base; beta-Lactams; cell growth; clinically relevant; clinically significant; combinatorial; density; experience; experimental study; improved; in vitro Model; in vivo; novel strategies; novel therapeutic intervention; pandemic disease; pathogenic bacteria; prevent; recurrent infection; resistant strain

Project Summary / Abstract Clinical uropathogenic Escherichia coli (UPEC) strains, which are the causative agent of most urinary tract infections (UTIs), form quiescent intracellular reservoirs in the host where they can evade the action of antibiotics. We recently found that many clinical UPEC isolates also enter a non-proliferative quiescent, but viable, state in vitro that is quorum-dependent and reversed by specific signals, including peptidoglycan fragments (1). Of the 145 UPEC strains tested, 51 entered quiescence in vitro (1), including the well-studied virulent strain CFT073 and the pandemic strain JJ1886, a group B2 multi antibiotic resistant ST131/H30-Rx blood isolate from a patient with fatal urosepsis. Here, we will determine if quiescent UPEC cells formed in vitro are capable of evading antibiotics across major antibiotic classes as a result of entering the quiescent state. Our in vitro model will lay the foundational groundwork to rapidly determine if a clinical UPEC isolate enters quiescence and if so which classes of antibiotics are effective at killing those quiescent cells. We propose to uncover the antibiotic tolerance features associated with quiescent UPEC and test strategies to induce proliferation, which may overcome antibiotic tolerance. In Aim 1, we will elucidate the bactericidal and bacteriostatic effects of major antibiotic classes on growing and quiescent cells of clinically significant UTIs, including JJ1886, a multidrug resistant ST131 strain, and CFT073, an ST73 strain. We will determine if antibiotic tolerance is reversed upon stimulating proliferation of quiescent cells by addition of external proliferation signals in vitro. In Aim 2, we will test if UPEC cells become quiescent upon invasion of cultured epithelial cells and if proliferants modify the antibiotic tolerance and susceptibility profiles of bacteria associated with intracellular reservoirs. SIGNIFICANCE: Most UTIs are caused by UPEC and many patients experience recurrent infections. The bacteria associated with recurrent infections are thought to survive antibiotic treatment by entering a quiescent state in bladder epithelial cells, which allows them to resume growth upon release into urine at a later time. The studies proposed here are straightforward yet they may have significant implications for treating recurrent UTIs. We will identify if quiescent UPEC cells have tolerances to specific classes of antibiotics, and susceptibility to others, whether this translates to the same or different tolerances and susceptibilities in intracellular reservoirs, and whether quiescent cell tolerance in intracellular reservoirs can be modified by addition of specific proliferants. Therefore, the studies proposed here may elucidate new therapeutic strategies to prevent both antibiotic tolerance in UPEC and recurrent UTIs caused by UPEC.

项目总结/摘要 临床尿路致病性大肠杆菌（UPEC）菌株，是大多数尿路疾病的病原体 感染（UTI），在宿主中形成静止的细胞内储库，在那里它们可以逃避 抗生素我们最近发现，许多临床UPEC分离株也进入非增殖性静止期， 活的，在体外处于群体依赖性状态，并被特定信号（包括肽聚糖）逆转 片段（1）。在测试的145株UPEC菌株中，51株在体外进入静止期（1），包括研究充分的 强毒株CFT 073和流行毒株JJ 1886，B2组多重抗生素耐药株ST 131/H30-Rx 从一个患有致命性尿脓毒症的病人身上分离出的血液在这里，我们将确定静止的UPEC细胞是否在体外形成， 由于进入静止状态，能够逃避主要抗生素类别的抗生素。 我们的体外模型将为快速确定临床UPEC分离株是否进入 如果是这样的话，哪类抗生素能有效地杀死那些静止的细胞。我们建议 揭示与静止期UPEC相关的抗生素耐受性特征和诱导UPEC的测试策略 增殖，这可能会克服抗生素耐受性。 在目标1中，我们将阐明主要抗生素类对细菌的杀菌和抑菌作用。 临床显著UTI的生长和静止细胞，包括JJ 1886，一种多药耐药ST 131 菌株，和ST 73菌株CFT 073。我们将确定抗生素耐药性是否会在刺激 通过体外加入外部增殖信号使静止细胞增殖。 在目标2中，我们将测试UPEC细胞在侵袭培养的上皮细胞后是否变得静止， 如果增殖物改变了与细菌相关的抗生素耐受性和敏感性， 细胞内储库 意义：大多数UTI是由UPEC引起的，许多患者经历复发性感染。的 与复发性感染相关的细菌被认为通过进入静止期而在抗生素治疗后存活， 在膀胱上皮细胞中的状态，这允许它们在稍后的时间释放到尿液中后恢复生长。的 这里提出的研究是直接的，但它们可能对治疗复发性UTI具有重要意义。 我们将确定静止的UPEC细胞是否对特定类别的抗生素具有耐受性，以及对抗生素的敏感性。 其他的，无论这是否转化为细胞内储存器中相同或不同的耐受性和相容性， 以及是否可以通过添加特异性的抗肿瘤药物来改变细胞内储库中的静止细胞耐受性。 扩散者。因此，这里提出的研究可能阐明新的治疗策略，以防止这两种疾病。 UPEC中的抗生素耐受性和由UPEC引起的复发性UTI。

项目成果

Metabolic flux regulates growth transitions and antibiotic tolerance in uropathogenic Escherichia coli.

代谢通量调节泌尿道致病性大肠杆菌的生长转变和抗生素耐受性。

• DOI: 10.1101/2023.05.09.540013
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Morrison,JosiahJ;Banas,DanielA;Madden,EllenK;DiBiasio,EricC;Rowley,DavidC;Cohen,PaulS;Camberg,JodiL
• 通讯作者: Camberg,JodiL