Copper Sensing in Uropathogenic Escherichia coli

尿路致病性大肠杆菌中的铜感应

• 批准号: 10604449
• 负责人: Seth A Reasoner
• 金额: $ 3.29万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604449
• 关键词: Acute; Address; Adult; Aerobic; Affect; Anatomy; Antibiotics; Antimicrobial Resistance; Attenuated; Bacteria; Bacterial Infections; Biochemical; Bladder; Chronic; Complement; Copper; Data; Defect; Development; Drug Metabolic Detoxication; Escherichia coli; Excretory function; Future; Genes; Genetic; Genetic Transcription; Growth; Homeostasis; Human; Immune response; Immunofluorescence Immunologic; Individual; Inductively Coupled Plasma Mass Spectrometry; Infection; Infective cystitis; Intestines; Knowledge; Laboratories; Mass Spectrum Analysis; Mediating; Membrane; Mentors; Metals; Methods; Microscopy; Modeling; Mus; Oxygen; Pathway interactions; Prevention; Prevention approach; Proteins; Recurrence; Regulation; Research Personnel; Role; Sampling; Shapes; Signal Transduction; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stress; System; Technical Expertise; Techniques; Testing; Therapeutic; Therapeutic Agents; Tissue Sample; Toxic effect; Tropism; Urinary tract; Urinary tract infection; Urine; Uropathogen; Uropathogenic E. coli; Vagina; Virulence; Work; acute infection; antimicrobial; antimicrobial drug; career; efflux pump; experience; fitness; genetic approach; genetic manipulation; gut colonization; host colonization; improved; insight; mass spectrometric imaging; mouse model; mutant; novel; novel therapeutics; oxidation; pathogen; pathogenic bacteria; periplasm; prevent; protein-histidine kinase; recurrent infection; respiratory; response; skills; spatiotemporal; therapeutic target; toxic metal; transcription factor

PROJECT SUMMARY Significance: Urinary tract infections (UTIs) are the most common adult bacterial infection annually affecting over 150 million individuals worldwide. As a common reason for antibiotic prescriptions, UTIs are considered a substantial source of resistance to antimicrobial agents. Thus, improved understanding of the interactions between host and bacterial pathogen during UTI is essential for facilitating the development of new therapeutic agents. The concentration of copper increases 5-fold in the urine during acute infection as a protective strategy against the pathogen. Most copper is excreted in the intestines where uropathogens colonize following UTI forming the basis for recurrent infections. Uropathogenic Escherichia coli (UPEC), the most common cause of UTIs, uses copper efflux and sequestration mechanisms to prevent copper toxicity. Additionally, UPEC inhabits multiple niches (intestines, vagina, and bladder) throughout the host that vary in copper content and oxygen concentration. While copper response mechanisms have been characterized in laboratory strains of E. coli, the relative contribution of each copper response system to UPEC persistence in the host has not been elucidated. This proposal will determine the spatio-temporal induction and regulation of the UPEC response to copper and will define the role of each response system in mediating UPEC persistence in the host. Rationale and Hypothesis: In order to successfully infect the urinary tract, UPEC must tolerate increased copper levels. Similarly, UPEC must tolerate intestinal copper excretion to colonize the host gut and potentially cause re-infections. Based on prior studies, the copper responsive UPEC locus cus is among the most upregulated of all genes in samples from acute human UTIs. The cus locus encodes the two-component signaling system CusSR, a copper sensing and response signaling system, and copper efflux apparatus CusCFBA. Additionally, the copper response systems CueR/CopA facilitate copper export in aerobic conditions. Oxygen availability is a key determinant of which copper response system is preferentially active in E. coli due to the oxidation state of copper and energetic requirements for copper efflux. Oxygen levels vary widely within UPEC’s niches. I hypothesize that oxygen availability and copper levels dictate the copper response systems that are active in the anatomic niches that UPEC colonizes and infects. I propose two aims to test this hypothesis: Aims: Aim 1 will determine which UPEC copper response systems are active in the anatomic niches (vagina, intestines, bladder) that UPEC encounters. Aim 2 will investigate the role of CusSR in mediating copper sensing and tolerance in the anaerobic intestinal niche. Impact: These studies will be the first to address the hierarchy and localization of copper response systems uropathogenesis. Clarification of how UPEC senses and responds to copper will provide insight into whether targeting copper detoxification methods of UPEC is a viable therapeutic option for UTI treatment and prevention.

项目摘要 意义：尿路感染（UTIs）是最常见的成人细菌感染，每年影响 全球超过1.5亿人。作为抗生素处方的常见原因，UTI被认为是一种 抗微生物剂耐药性的主要来源。因此，更好地理解了 在UTI期间宿主和细菌病原体之间的相互作用对于促进新治疗药物的开发是至关重要的。 剂.在急性感染期间，尿中铜的浓度增加5倍，作为一种保护性策略 抵抗病原体大多数铜在尿路感染后尿病原体定植的肠道中排泄 形成反复感染的基础。尿路致病性大肠杆菌（UPEC），最常见的原因， UTIs使用铜流出和螯合机制来防止铜毒性。此外，UPEC栖息在 多个壁龛（肠，阴道和膀胱）在整个主机不同的铜含量和氧气 浓度.虽然铜反应机制已在实验室菌株的E。杆菌 各铜响应系统对宿主中UPEC持久性的相对贡献尚未阐明。 该建议将确定UPEC对铜的响应的时空诱导和调节， 将定义每个响应系统在介导宿主中的UPEC持久性中的作用。 原理和假设：为了成功感染尿路，UPEC必须耐受增加的 铜水平。类似地，UPEC必须耐受肠铜排泄以定殖宿主肠道，并可能 导致再次感染。根据先前的研究，铜响应性UPEC位点cus是其中最重要的 在来自急性人UTI的样品中所有基因都上调。cus基因座编码双组分 信号系统CusSR、铜传感和响应信号系统以及铜流出装置 CusCFBA。此外，铜响应系统CueR/CopA促进铜在有氧条件下输出。 氧的可利用性是决定大肠杆菌中哪种铜反应系统优先活性的关键因素。大肠杆菌 铜的氧化态和铜流出的能量要求。体内的氧气含量变化很大 UPEC的壁龛。我假设氧气的可用性和铜的水平决定了铜的反应 在UPEC定植和感染的解剖学小生境中活跃的系统。我提出两个目标来检验 这个假设： 目的：目的1将确定哪些UPEC铜反应系统在解剖学小生境（阴道， 肠，膀胱），UPEC遇到的。目的2将研究CusSR在介导铜传感中的作用 和耐受性。 影响：这些研究将是第一个解决铜反应系统的层次和定位的研究。 泌尿系发病机制阐明UPEC如何感知和响应铜将提供洞察， UPEC的靶向铜解毒方法是UTI治疗和预防的可行治疗选择。