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.

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