Mechanistic basis of urinary Lactobacillus enrichment by estrogen hormone therapy

提案摘要/摘要 泌尿道感染（UTI）的医疗负担很大，因为它们直接占所有临床疾病的1%。 访问美国。目前，UTI每年的医疗费用超过35亿美元。作为全球 从健康角度来看，UTI主要影响女性，当患者 在12个月内经历3次或以上UTI。鲁蒂的风险随着年龄的增长而增加， 绝经后（PM）妇女的发病率不成比例地高。鲁蒂的有效疗法严重 有限的，因为大多数治疗策略依赖于长期抗生素治疗，目标是在 泌尿道然而，即使是健康的尿液也不是无菌的，已经证实， 存在于PM女性的健康泌尿道中，这里称为尿生物群系。迄今为止，大多数治疗策略 对于鲁蒂，不考虑保留或恢复泌尿或泌尿生殖道的常驻微生物群 道。有趣的是，已经显示尿生物群系与阴道微生物群系和阴道微生物群系相互关联。 生产D（-）乳酸的乳酸杆菌，如卷曲乳酸杆菌，已被证明可以防止细菌感染。 阴道病阴道乳酸杆菌已与循环雌激素水平和早期临床研究 发现雌激素治疗（EHT）与PM中阴道乳酸杆菌富集密切相关 妇女有趣的是，EHT现在被广泛认为是PM中鲁蒂的有效治疗选择 妇女，但这一观察背后的机制是未知的。我们观察到一种强烈的关联 使用EHT和尿L之间的关系。在我们的宏基因组分析中， PM女性的对照横断面队列（n=75）。这些观察使我提出一个基本的问题， 问题：雌激素如何富集尿生物群中的乳杆菌群落？该提案以 EHT作为鲁蒂的治疗方法，增强了大肠杆菌在大肠杆菌中的定植。 通过刺激宿主产生所需的营养源和支架附着位点 为了殖民。为了验证这些假设，我将描述EHT治疗对代谢的影响， 培养原代尿路上皮细胞，并使用来自我们的去识别人类的细菌分离物的策划生物库， 研究尿路上皮细胞和L.在存在和不存在 雌激素.该提案的长期目标是利用从测试这些假设中获得的见解， 确定鲁蒂有效和安全辅助治疗的替代治疗靶点。这其中的一个中心主题是 我和我的赞助者认为，我的建议是利用科学目标作为实现关键训练目标的工具， 这对我未来的独立研究有很大的帮助。

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.