Conserved Proteus mirabilis genetic requirements for colonization of the catheterized urinary tract

导尿管定植的保守奇异变形杆菌遗传要求

• 批准号: 10417123
• 负责人: Chelsie Elizabeth Armbruster
• 金额: $ 34.89万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10417123
• 关键词: Address; Amino Acids; Animal Model; Antibiotic Resistance; Antibiotic Therapy; Bacteremia; Bacteria; Bladder; Blood Circulation; Catheterization; Catheters; Cells; Cessation of life; Clinical; Data; Defense Mechanisms; Development; Disease; Drug Metabolic Detoxication; Drug resistance; Enterococcus; Enterococcus faecalis; Enzymes; Escherichia coli; Exposure to; Extended-spectrum β-lactamase; Future; Genes; Genetic; Genome; Gram-Negative Bacteria; Growth; Health; Hospitals; Human; Hydrogen Peroxide; Immune; In Vitro; Indwelling Catheter; Infection; Infection prevention; Inflammation; Intervention; Invaded; Kidney; Knowledge; Length of Stay; Libraries; Life; Measures; Mediating; Metabolic Pathway; Metabolism; Mission; Modeling; Morganella morganii; Multi-Drug Resistance; Mus; Organism; Oxidative Stress; Pathogenicity; Pathway interactions; Patients; Peroxides; Physiological; Prevention; Process; Production; Proteus mirabilis; Providencia; Reactive Oxygen Species; Reporting; Research; Respiratory Burst; Risk; Role; Sampling; Serine; Site; Testing; Therapeutic Intervention; United States National Institutes of Health; Urinary Calculi; Urinary tract; Urine; Uropathogen; Vaccines; Validation; adverse outcome; alkyl hydroperoxide reductase; amino acid metabolism; antimicrobial; bacterial fitness; base; carbapenemase; catalase; catheter associated UTI; clinically relevant; co-infection; disorder prevention; drug resistant bacteria; effective therapy; fitness; functional decline; genome-wide; health care settings; healthcare-associated infections; in vivo; interest; metabolic fitness; mortality; mouse model; multi-drug resistant pathogen; mutant; neutrophil; novel; novel strategies; preference; prevent; renal damage; response; urinary

Project Summary/Abstract Patients with indwelling urinary catheters have nearly double the mortality rate compared to non-catheterized patients due to the propensity for adverse outcomes, including functional decline, increased length of stay in hospital settings, catheter-associated urinary tract infections (CAUTI), and bacteremia. Catheterization also increases the likelihood of colonization by multidrug-resistant organisms, and antibiotic resistance is increasing at an alarming rate, making it imperative to identify novel, non-antibiotic treatments for CAUTI and associated complications. The Gram-negative bacterium Proteus mirabilis is a predominant cause of CAUTI, particularly during long-term catheterization. CAUTI is also frequently polymicrobial, which can increase risk of severe disease and bacteremia. There is, however, a fundamental gap in knowledge regarding conserved targets for treating or preventing disease due to P. mirabilis, particularly during polymicrobial infection. Our preliminary data clearly demonstrates that the presence of other bacterial species during infection dramatically impacts the genes required by P. mirabilis for colonization and persistence during CAUTI, especially the metabolic pathways that are favored by the bacterium for growth and the defense mechanisms that are used to evade host antimicrobial responses. However, we have uncovered a set of 217 genes encoded by P. mirabilis that provide a fitness advantage during single-species infection and polymicrobial infection with another Gram-negative bacterium, Providencia stuartii. Fifty-seven percent of these genes (123/217) are highly conserved in all 106 publically- available P. mirabilis genomes. The central hypothesis of this proposal is that P. mirabilis encodes a core set of conserved genes that are critical for establishing CAUTI, regardless of which other bacterial species are present. We further anticipate that a subset of these factors will be ideal targets for treatment or prevention of disease. This hypothesis will be tested through three concurrent specific aims. In Aim 1, we will utilize genome- wide transposon insertion-site sequencing to uncover the impact of co-colonization by two common uropathogens (Escherichia coli and Enterococcus faecalis) on the genes required by P. mirabilis to colonize the catheterized urinary tract, including which of the 123 conserved core fitness factors remain important for establishing infection and whether they are expressed during human CAUTI. In Aims 2 and 3, we will explore the importance of two pathways that are well-represented in the current set of core fitness factors (amino acid metabolism and peroxide detoxification) to P. mirabilis pathogenicity. This includes broad characterization of amino acid availability within the murine urinary tract, with or without an indwelling catheter, and the amino acid preferences of 5 uropathogens, as well as production of reactive oxygen species by these uropathogens and by neutrophils in response to the uropathogens. The knowledge gained herein will uncover unique challenges that the catheterized urinary tract presents to invading uropathogens, as well as novel, conserved, clinically-relevant targets for treating or preventing infections due to these prevalent and increasingly drug-resistant bacteria.

项目概要/摘要 留置导尿管患者的死亡率几乎是未留置导尿管患者的两倍 患者有出现不良后果的倾向，包括功能下降、住院时间延长 医院环境、导管相关尿路感染 (CAUTI) 和菌血症。还进行导尿术 增加了多重耐药微生物定植的可能性，并且抗生素耐药性正在增加 正以惊人的速度发展，因此迫切需要寻找新的、非抗生素的治疗方法来治疗 CAUTI 和相关疾病。 并发症。革兰氏阴性菌奇异变形杆菌是 CAUTI 的主要原因，尤其是 长期导尿期间。 CAUTI 也常常是多种微生物，这会增加严重的风险 疾病和菌血症。然而，对于保守目标的认识存在根本差距。 治疗或预防由奇异变形杆菌引起的疾病，特别是在多种微生物感染期间。我们的初步数据 清楚地表明感染过程中其他细菌种类的存在会极大地影响基因 P. mirabilis 在 CAUTI 期间定植和持续存在所需，特别是代谢途径 受到细菌生长和用于逃避宿主抗菌药物的防御机制的青睐 回应。然而，我们发现了由 P. mirabilis 编码的一组 217 个基因，这些基因提供了适应性 在单种感染和另一种革兰氏阴性菌的多种微生物感染中具有优势， 斯图亚特普罗维登斯。这些基因中的 57% (123/217) 在所有 106 个公开的基因中高度保守—— 可用的奇异果基因组。该提案的中心假设是奇异果编码了一个核心集 对于建立 CAUTI 至关重要的保守基因，无论其他细菌种类如何 展示。我们进一步预计这些因素的一部分将成为治疗或预防疾病的理想目标 疾病。这一假设将通过三个同时存在的具体目标进行检验。在目标 1 中，我们将利用基因组 广泛的转座子插入位点测序揭示了两种常见转座子共定植的影响 尿路病原体（大肠杆菌和粪肠球菌）对奇异变形杆菌定植所需基因的影响 插管尿路，包括 123 个保守的核心健康因素中的哪些对于 建立感染以及它们是否在人类 CAUTI 期间表达。在目标 2 和 3 中，我们将探索 当前一组核心健康因子（氨基酸）中充分体现了两条途径的重要性 代谢和过氧化物解毒）对奇异果的致病性。这包括广泛的特征 小鼠泌尿道内氨基酸的可用性，有或没有留置导管，以及氨基酸 5 种尿路病原体的偏好，以及这些尿路病原体和活性氧的产生 中性粒细胞对尿路病原体的反应。在此获得的知识将揭示独特的挑战 插管尿路会出现入侵的尿路病原体，以及新颖的、保守的、临床相关的 治疗或预防这些普遍且耐药性日益增强的细菌引起的感染的目标。