Investigating metabolism and DNA damage repair in uropathogenic Escherichia coli fluoroquinolone persisters

研究泌尿道致病性大肠杆菌氟喹诺酮类持续存在的代谢和 DNA 损伤修复

• 批准号: 10747651
• 负责人: TRAVIS LAGREE
• 金额: $ 4.34万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-08 至 2026-09-07
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10747651
• 关键词: Address; Adjuvant; Affect; Aftercare; Anabolism; Antibiotic Therapy; Antibiotics; Bacteria; Bacterial Infections; Bacterial Physiology; Biological Assay; Carbon; Cell Survival; Cells; Charge; Chemicals; Chemistry; Chemosensitization; Chimeric Proteins; Clinical; Complement; DNA; DNA Binding; DNA Damage; DNA Repair; DNA Repair Gene; DNA Topoisomerases; DNA biosynthesis; DNA lesion; Data; Development; Environment; Escherichia coli; Escherichia coli Infections; Event; Excision; Flow Cytometry; Fluorescence; Fluorescence Microscopy; Fluorescence-Activated Cell Sorting; Fluorescent Antibody Technique; Fluorescent Dyes; Fluoroquinolones; Frequencies; Future; Gene Expression; Genes; Genetic; Genetic Transcription; Glucose; Growth; Infection; Knowledge; Mass Spectrum Analysis; Measures; Metabolic; Metabolism; Microscopy; Modeling; Molecular; Mutation; Nucleic Acids; Nucleotides; Nutrient; Nutrient availability; Persons; Pharmaceutical Preparations; Phenotype; Physiology; Population; Predisposition; RNA; Recovery; Recurrence; Relapse; Reporter; Research; Resistance; Resistance development; Sorting; Source; Stress; System; Testing; Topoisomerase; Translating; Treatment Failure; Urinary tract; Urinary tract infection; Urine; Uropathogenic E. coli; adenylate; antibiotic tolerance; bacterial metabolism; bactericide; differential expression; drug resistant pathogen; experimental study; genetic approach; improved; insight; metabolomics; microbial disease; mutant; non-genetic; nutrient deprivation; persistent bacteria; programs; repaired; response; success; transcriptome sequencing; treatment response

PROJECT SUMMARY: Uropathogenic Escherichia coli (UPEC) is one of the major causative agents of urinary tract infections (UTIs). With growing frequency of antibiotic treatment failure and slowing of antibiotic discovery, the rate of recurrent UTIs is poised to continue to increase. Although antibiotic resistant pathogens are of particular concern, treatment failure can often occur by non-genetic mechanisms, without detectable resistance. One of the non- genetic mechanisms of antibiotic treatment failure—persistence—is characterized by antibiotic tolerance in a small group of cells among a susceptible population. Bacterial persisters, which can reversibly exit the antibiotic- tolerant state following drug removal, is able to repopulate an infection and impede success of treatment. Persisters are known to be enriched in slow growing populations, where overall cellular activity and metabolism is reduced; however, there is a lack of knowledge on how the surrounding nutrient environment can affect bacterial physiology, and how these factors affect susceptibility to antibiotics. Our central objective is to determine the impact of carbon source availability on UPEC persistence to fluoroquinolone (FQ) antibiotics, which are bactericidal through the targeting of DNA topoisomerases and the accumulation of DNA damage. Aim 1 aims to address changes to metabolism, biomolecular synthesis, and DNA integrity in response to carbon source availability during FQ treatment. We will assess these factors by performing experiments with fluorescence-based probes, as well as mass spectrometry analysis of metabolites. These data will elucidate the ways by which carbon source availability can impact the susceptibility of cells to FQs, allowing us to explore specific mechanisms as potentiation targets. Aim 2 will enable us to understand how the coordination of DNA damage response changes due to carbon source availability after FQ treatment. We will deploy powerful genetic approaches and RNA sequencing in E. coli to interrogate the timing of gene expression and DNA repair. Completion of this aims will provide great insight into the importance of various DNA repair mechanisms in response to FQ treatment. Overall, the completion of these aims will reveal metabolic, biosynthetic, and expressional changes that underlie FQ persistence in E. coli. This information could illuminate potential genes that can be targeted to potentiate the activity of FQs and reduce resistance development in persister progenies. Additionally, these findings could lead to interesting implications for clinical infections, with a potential connection between host metabolism and antibiotic efficacy. This research could highlight the potential of adding glucose, as well as other metabolites, as supplemented compounds during antibiotic administration to improve the treatment of infections.

项目摘要： 尿道病大肠杆菌（UPEC）是尿路感染（UTI）的主要结构药物之一。 随着抗生素治疗衰竭频率的增长和抗生素发现减慢的频率，复发率 UTI被中毒以继续增加。尽管抗生素耐药病原体特别关注 通常没有可检测的耐药性，通常会通过非遗传机制发生治疗失败。非 - 抗生素治疗衰竭的遗传机制 - 垂直性 - 以抗生素耐受性为特征 易感人群中的小组细胞。细菌持久性可以可逆地退出抗生素 - 去除药物后的耐受状态，能够重新填充感染并阻碍治疗的成功。 众所周知，在整体细胞活性和代谢中，持久者富集了人群中的慢速人群 减少；但是，缺乏关于周围营养环境如何影响的知识 细菌生理学以及这些因素如何影响抗生素的易感性。 我们的核心目的是确定碳源可用性对UPEC持久性的影响 氟喹诺酮（FQ）抗生素，通过靶向DNA拓扑异构酶和 DNA损伤的积累。 AIM 1的目的是解决对代谢，生物分子合成和DNA的变化 响应FQ处理期间碳源可用性的完整性。我们将通过 对基于荧光的问题进行实验，以及代谢物的质谱分析。 这些数据将阐明碳源可用性可以影响细胞的敏感性的方式 FQ，使我们能够探索特定机制作为潜在目标。 AIM 2将使我们能够了解如何 FQ处理后碳源可用性导致的DNA损伤响应的协调变化。我们 将在大肠杆菌中部署强大的遗传方法和RNA测序以询问基因的时间 表达和DNA修复。完成此目标的完成将为各种DNA的重要性提供深刻的了解 响应FQ治疗的修复机制。 总体而言，这些目标的完成将揭示代谢，生物合成和表达变化，这是基础的 大肠杆菌中的FQ持久性。这些信息可以照亮可以针对潜在的潜在基因 FQ的活性并降低持久后代的抵抗力发展。此外，这些发现可能会带来 对临床感染的有趣含义，宿主代谢和 抗生素效率。这项研究可能突出添加葡萄糖以及其他代谢产物的潜力 补充抗生素给药期间的化合物以改善感染的治疗。