Mechanistic basis of urinary Lactobacillus enrichment by estrogen hormone therapy

建议书摘要/摘要 尿路感染(UTI)的医疗负担很大，因为它们直接占临床总人数的1% 在美国的访问。目前，UTI每年的医疗费用超过35亿美元。作为全球 健康问题，尿路感染主要影响女性，通常进展为复发性尿路感染(RUTI) 在12个月内经历3次或3次以上尿路感染。Ruti的风险随着年龄的增长而增加，导致 绝经后(PM)妇女的发病率高得不成比例。鲁蒂的有效治疗方法是严重的 有限，因为大多数治疗策略依赖于长期的抗生素治疗，目标是在 尿路。然而，即使是健康的尿液也不是无菌的，而且已经证实，微生物群落 驻留在PM妇女的健康尿路中，在这里被称为尿菌群。到目前为止，大多数治疗策略 对于Ruti来说，不考虑保留或恢复尿路或泌尿生殖系统的驻留微生物区系 一条小路。有趣的是，尿菌群已被证明与阴道微生物群和阴道相互联系。 生产D(-)乳酸的乳杆菌，如皱纹乳杆菌，已被证明可以保护细菌免受细菌的侵害。 阴道炎。阴道乳杆菌与循环雌激素水平和早期临床研究有关 发现雌激素治疗(EHT)与PM的阴道乳酸菌增多密切相关 女人。有趣的是，EHT现在被广泛认为是PM中Ruti的有效治疗选择 但这种观察背后的机制尚不清楚。我们观察到了一种强烈的联系 EHT的使用和尿中皱纹乳杆菌在我们的尿液生物组学分析中的应用 PM妇女对照横断面队列(n=75)。这些观察结果让我问了一个基本的问题 问：雌激素如何丰富泌尿生物群中的乳杆菌群落？这份提案的核心是 围绕以下假设：EHT作为治疗RUTI的一种方法，增强了乳杆菌在 通过刺激宿主产生营养来源和支架所需的附着部位来实现尿路 为了殖民统治。为了验证这些假说，我将描述EHT治疗对 培养原代尿路上皮细胞，并使用来自我们未鉴定的人的细菌分离的精选生物库 研究有无尿路上皮细胞与皱纹乳杆菌共培养定植的队列 雌激素。这项提议的长期目标是利用从测试这些假设中获得的洞察力来 为Ruti的有效和安全的辅助治疗确定替代治疗靶点。这件事的一个中心主题 建议将科学目标作为实现关键培训目标的工具，我和我的赞助商相信 对我未来的独立研究事业有很大的帮助。

Proposal Summary/Abstract The medical burden of urinary tract infection (UTI) is significant as they directly contribute to 1% of all clinical visits in the United States. Annual medical expenses attributed to UTI currently exceed $3.5 billion. As a global health concern, UTIs mainly affect women and commonly progress into recurrent UTI (rUTI) when a patient experiences 3 or more UTIs in a 12 month period. The risk of rUTI increases with age, leading to disproportionately high incidence rates in postmenopausal (PM) women. Effective therapies for rUTI are severely limited as most treatment strategies rely on long-term antibiotic therapy with the goal of achieving sterility within the urinary tract. However, even healthy urine is not sterile, and it has been confirmed that a microbial community resides in the healthy urinary tract of PM women, termed here the urobiome. To date, most therapeutic strategies for rUTI do not consider the preservation or restoration of the resident microbiota of the urinary or urogenital tract. Interestingly, the urobiome has been shown to be interconnected with the vaginal microbiome and vaginal D(-)Lactate-producing lactobacilli, such as Lactobacillus crispatus, have been shown to protect against bacterial vaginosis. Vaginal lactobacilli have been associated with circulating estrogen levels and early clinical studies found that estrogen hormone therapy (EHT) is strongly associated with vaginal lactobacilli enrichment in PM women. Interestingly, EHT is now widely becoming recognized as an effective treatment option for rUTI in PM women, but the mechanism behind this observation is unknown. We have observed a strong association between EHT use and urinary L. crispatus enrichment in our metagenomic analysis of the urobiome of a controlled cross-sectional cohort of PM women (n=75). These observations have led me to ask a fundamental question: How does estrogen enrich for Lactobacillus communities in the urobiome? This proposal centers around the following hypothesis: EHT, as a treatment for rUTI, enhances Lactobacillus colonization of the urinary tract by stimulating host production of nutrient sources and scaffolding attachment sites needed for colonization. To test these hypotheses, I will characterize the effect of EHT treatment on the metabolism of cultured primary urothelial cells and use a curated biobank of bacterial isolates from our de-identified human cohorts to study colonization of co-cultures of urothelial cells and L. crispatus in the presence and absence of estrogen. The long-term goal of this proposal is to leverage the insight gained from testing these hypotheses to identify alternate therapeutic targets for effective and safe adjunct therapies for rUTI. A central theme of this proposal is to use the scientific goals as a vehicle to achieve critical training goals I and my sponsors believe will greatly help in my future independent research career.