Estrogenic Modulation of Genitourinary Epithelial-Probiotic Cross-Talk

描述(申请人提供)：泌尿生殖道感染影响全球儿童和成人。肠道来源、性传播或垂直传播的病原体通过阴道传播，感染女性生殖道。然而，共生乳杆菌生物膜可能通过调节参与自然杀伤T(NKT)细胞活性的基因，如干扰素-γ(IFNG)受体1和细菌脂质抗原递呈分子CD1d，来启动泌尿生殖道上皮细胞的抗致病反应。反过来，阴道乳杆菌生物膜的形成可能依赖于雌激素诱导的上皮糖原的产生，糖原是共生菌衍生的益生菌的食物来源，“活的微生物，当给予足够的量时，会给宿主带来健康益处”。我们假设雌激素对泌尿生殖道上皮细胞糖原产生的刺激促进了益生菌乳杆菌生物膜的稳定性，该生物膜相互调节上皮CD1d介导的IFNG相关的NKT细胞对病原体的反应。这一假说将通过以下目的得到验证：目的1，阐明雌激素诱导的糖原对与生殖道上皮细胞相关的生物膜的影响；目的2，研究益生菌介导的NKT细胞与生殖道上皮相互作用的体内和体外机制；以及目的3，阐明雌激素、生殖道上皮和NKT细胞相互作用在形成生殖道微生物群中的微生态作用。通过这些研究，候选人寻求成为一名内科研究员，通过培训生物膜微生物学和天然免疫学来补充他在儿科泌尿科和适应性免疫学方面的研究生工作背景。在候选人职业生涯的早期阶段，候选人将通过以下方式建立他作为内科科学家的基础：技能研讨会；生物膜生物学、先天免疫和儿科泌尿外科专家的强大指导；科学合作；实践经验；以及专注于生物膜、先天免疫和生殖生物学的教学课程。总而言之，该项目将为成为一名成功的独立调查员奠定基础，并将导致确定雌激素和益生菌保护女性生殖道免受感染的新途径。 公共卫生相关性：共生阴道细菌的生物膜可能具有益生菌(有益)特性，使生殖道为自然杀伤T细胞对感染的反应做好准备。这项提案将研究这些生物膜是如何在生殖道上皮细胞上生长的，以及益生菌、雌激素、生殖道上皮细胞和自然杀伤T细胞如何相互作用来确定哪些细菌在女性生殖道中生长。

DESCRIPTION (provided by applicant): Genitourinary infections affect children and adults worldwide. Gut-derived or sexually or vertically transmitted pathogens travel through the vagina to infect the female genitourinary tract. However, commensal Lactobacillus biofilms may prime genitourinary epithelial cells for anti-pathogenic responses by regulating genes which participate in natural killer T (NKT) cell activity, such as interferon gamma (IFNG) receptor 1 and the bacterial lipid antigen-presenting molecule CD1d. In turn, the formation of vaginal Lactobacillus biofilms may depend on estrogen-induced, epithelial production of glycogen, a food source for commensal-derived probiotics, "live microorganisms which when administered in adequate amounts confer a health benefit on the host". We hypothesize that estrogen stimulation of glycogen production by genitourinary epithelium promotes stability of probiotic Lactobacillus biofilms, which reciprocally modulate epithelial CD1d-mediated, IFNG-associated NKT cell responses to pathogens. This hypothesis will be tested through the following aims: Aim 1, elucidate the effects of estrogen-induced glycogen on biofilms associated with genitourinary epithelial cells; Aim 2, investigate the in vivo and in vitro mechanisms of probiotic-mediated, NKT cell interactions with genitourinary epithelium; and Aim 3, clarify the microecologic role of estrogen, genitourinary epithelial, and NKT cell interactions in shaping the genitourinary microbiome. Through these studies, the candidate seeks to become a physician investigator by complementing his background in pediatric urology and graduate work in adaptive immunology with training in biofilm microbiology and innate immunology. During the early portion of the candidate's career, the candidate will build on his foundation as a physician scientist through grantsmanship workshops; strong mentorship by experts in biofilm biology, innate immunity, and pediatric urology; scientific collaborations; practical experience; and didactic coursework that focuses on biofilms, innate immunity, and genitourinary biology. In summary, this project will lay the groundwork for a successful career as an independent investigator and will lead to the identification of novel pathways by which estrogen and probiotics protect the female genitourinary tract from infections. PUBLIC HEALTH RELEVANCE: Biofilms of commensal vaginal bacteria may have probiotic (beneficial) properties which prime the genitourinary tract for natural killer T cell responses against infections. This proposal will examine how these biofilms grow on genitourinary epithelium, and how probiotics, estrogen, genitourinary epithelium, and natural killer T cells interact to determine which bacteria grow in the female genitourinary tract.