Quelling UPEC Proliferation by Lactobacillus Mediated Induction of Latent Phages

通过乳酸菌介导的潜伏噬菌体诱导抑制 UPEC 增殖

• 批准号: 10514959
• 负责人: Swarnali Banerjee
• 金额: $ 41.09万
• 依托单位: LOYOLA UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-10 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10514959
• 关键词: Academic Research Enhancement Awards; Acute; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Genome; Bacteriophages; Benign; Bioinformatics; Biology; Bladder; Catalogs; Cells; Cessation of life; Chicago; Coculture Techniques; Coliphages; Collection; Communities; Cytolysis; Development; Environment; Escherichia coli; Female; Frequencies; Generations; Genes; Genome; Genomics; Genotype; Growth; Human; Hydrogen Peroxide; Institution; Interdisciplinary Study; Investigation; Kidney; Lactic acid; Lactobacillus; Life Cycle Stages; Light; Liquid substance; Lytic; Measures; Mediating; Methods; Microbe; Microbiological Techniques; Play; Population; Process; Production; Protocols documentation; RNA Interference; Research; Research Personnel; Role; Sampling; Serotyping; Shapes; Stress; Surveys; Taxonomy; Testing; Time; United States National Institutes of Health; Universities; Urinary Microbiome; Urinary tract; Urinary tract infection; Urine; Urologic Diseases; Uropathogenic E. coli; Vagina; Work; acute care; biological adaptation to stress; clinical care; design; experimental study; interdisciplinary approach; lower urinary tract symptoms; member; microbiota; statistics; stressor; undergraduate student; urinary; virome

Prior studies have found that several Lactobacillus species inhibit the growth of or kill uropathogenic Escherichia coli (UPEC). Lactobacillus crispatus, L. gasseri, L. iners, and L. jensenii are frequent dominant members of both the healthy female vaginal and urinary microbiota and have been associated with the lack of lower urinary tract symptoms, including urinary tract infections (UTIs). Urinary Lactobacilli produce hydrogen peroxide (H2O2) and lactic acid, both of which can stress the UPEC cell and are attributed to reduced colonization within the bladder. As phage biologists and the first group to investigate phages within the urobiome, we recognized an immediate connection between Lactobacillus-dominated environments and temperate phage induction. Our prior genomic survey of bacteria in the urinary tract (urobiome) found that most bacterial genomes have latent (integrated) phage genomes, and our prior empirical work suggests that most of these latent phages are in fact temperate phages capable of being induced. The induced phage excises from the bacterial genome, replicates, and lyses (kills) its bacterial host cell. Our proof-of-concept work found: (i) cell-free Lactobacillus supernatant can induce UPEC latent temperate phages (coliphages), (ii) liquid cultures at Lactobacillus-relevant pH values can induce UPEC temperate coliphages, and (iii) liquid cultures at Lactobacillus-relevant H2O2 levels can induce UPEC temperate coliphages. This leads to our hypothesis: Lactobacilli may control UPEC colonization via “lysis from within” – H2O2 and/or pH stress UPEC strains, which leads to induction of resident temperate coliphages and death of the UPEC cell. Taking an interdisciplinary approach, the proposed project will assess the frequency and cause of induction of UPEC coliphages by Lactobacillus. Aim 1 will expand our preliminary work to determine if the cell-free supernatant of all common urinary Lactobacillus species induces UPEC coliphages; multiple strains of L. crispatus, L. gasseri, L. iners, and L. jensenii will be tested on a large collection of E. coli isolated from the bladders of females with and without LUTS, including UTI. In Aim 2, we will ascertain if phage induction triggered by cell-free Lactobacillus supernatant is the result of stress triggered by pH and/or H2O2 levels. Additionally, we will determine how pH and H2O2 impact UPEC growth over time. Our investigation will determine what role phage induction plays in shaping the urobiome community. Understanding the mechanisms that control UPEC colonization can inform clinical care of acute UTIs. This project is proposed as an R15 AREA for Undergraduate-Focused Institutions and is designed to engage undergraduates in interdisciplinary research of benign urologic disease.

先前的研究已经发现，几种乳杆菌抑制或杀死 尿路致病性大肠杆菌（UPEC）。卷曲乳杆菌Lactobacillus crispatus，L.加塞里湖iners和L. jensenii是健康女性阴道和泌尿微生物群的常见优势成员 并与缺乏下尿路症状有关，包括泌尿道 感染（UTI）。尿乳酸杆菌产生过氧化氢（H2 O2）和乳酸，两者均 其可压迫UPEC细胞并归因于膀胱内的减少的定殖。作为 噬菌体生物学家和第一个研究尿生物组内的噬菌体的小组，我们认识到， 乳酸杆菌优势环境与温带噬菌体之间直接联系 诱导我们先前对尿路细菌（尿生物群系）的基因组调查发现， 细菌基因组具有潜伏的（整合的）噬菌体基因组，我们先前的实验工作表明 大多数潜在的焦虑实际上是温和的焦虑，是可以被诱发的。的 诱导噬菌体从细菌基因组中切除，复制并裂解（杀死）其细菌宿主 cell.我们的概念验证工作发现：（i）无细胞乳酸菌上清液可以诱导UPEC 潜伏性温和细菌（大肠杆菌），（ii）乳酸杆菌相关pH值下的液体培养物可 诱导UPEC温和大肠杆菌，和（iii）乳酸杆菌相关H2 O2水平的液体培养物 可诱导UPEC温和大肠杆菌噬菌体。这导致我们的假设：乳酸杆菌可能控制 通过“从内部裂解”-H2 O2和/或pH应激UPEC菌株的UPEC定殖， 导致诱导定居性温带大肠杆菌和UPEC细胞死亡。 拟议的项目将采用跨学科的方法， 乳酸菌诱导UPEC大肠杆菌的原因。目标1将扩大我们的前期工作 为了确定所有常见的尿乳杆菌属物种的无细胞上清液是否诱导 UPEC大肠杆菌; crispatus，L.加塞里湖iners和L.詹森尼将接受测试 大量的E.从患有和不患有LUTS的女性膀胱中分离的大肠杆菌， 包括UTI。在目标2中，我们将确定是否由无细胞乳酸杆菌触发噬菌体诱导 上清液是由pH和/或H2 O2水平触发的应激的结果。此外，我们将 确定pH和H2 O2如何影响UPEC随时间的生长。我们的调查将决定 噬菌体诱导在形成尿生物群系中的作用。了解机制 控制UPEC定植可以为急性UTI的临床护理提供信息。 该项目被提议作为面向本科院校的R15区域， 旨在吸引本科生参与良性泌尿系统疾病的跨学科研究。

项目成果

Draft genomes of Lactobacillus jensenii UMB0908, UMB1545, and UMB5777 from the urinary tract.

来自泌尿道的詹氏乳杆菌 UMB0908、UMB1545 和 UMB5777 的基因组草图。

• DOI: 10.1128/mra.01120-23
• 发表时间: 2024
• 期刊: Microbiology resource announcements
• 影响因子: 0.8
• 作者: Steiling,Maria;Jackson,Briana;Stegman,Natalie;Sedano,Cerena;Putonti,Catherine
• 通讯作者: Putonti,Catherine