Methods to Test the Role of Age-related Lifestyle and Vaginal Microenvironment Changes and the Prevention, Treatment, and Progression of Genitourinary Syndrome of Menopause

超过50%的绝经后妇女受到绝经期泌尿生殖系统综合征（GSM）的影响， 与雌激素下降相关的进行性慢性疾病，包括阴道萎缩，阴道干燥， 性欲下降，以及其他症状和体征。GSM症状是一个代价高昂的公共卫生问题， 如果不治疗会恶化。绝经期雌激素下降被认为会导致阴道分泌物减少， 糖原和阴道微生物群的变化。这些包括低阴道乳杆菌属。50%至80%的 绝经后妇女。乳杆菌属部分通过乳酸保护泌尿生殖道免受病原体的侵害 生产和假设的抗炎特性，但是否乳酸杆菌发挥功能性作用， GSM未知外阴阴道GSM症状的可用治疗方法有局限性。有些女性 禁忌激素治疗或担心副作用。虽然阴道润滑剂提供 有些缓解，许多是有毒的阴道上皮，减少乳酸杆菌，并提高泌尿生殖道感染的风险。 因此，需要治疗和预防GSM的新方法。因为乳酸杆菌在绝经期经常减少， 阴道微生物群是合理的治疗目标;然而，单独的益生菌尚未被证明有效。 此外，阴道微生物群中与年龄相关的变化与代谢物和免疫特征共同变化;但是， 这些分子特征和它们在GSM中的作用之间的讨论是未知的。我们假设阴道中心 微环境生物标志物（VMB;例如，微生物、代谢物和免疫谱）反映阴道生物学特性 衰老（V-BA），在更年期增加，但可能是由生活方式因素改变。前瞻性研究 需要识别与年龄相关的核心VMB并确定它们如何影响GSM。为了实现这一目标，我们将 利用从812名年龄在35 - 60岁之间的妇女中收集的2，301份存档的宫颈阴道样本， 每6个月访视一次，持续2年（R01-CA 123467）。具体目的是1）使用VMB定量V-BA; 2） 纵向评价V-BA与GSM之间的关系; 3）纵向评价V-BA与GSM之间的关系。 生活方式因素对V-BA和VMB的影响; 4）量化V-BA和VMB在生活方式和 全球通。已经通过16S rRNA基因扩增子测序（R21-AI107224）对微生物群进行了分析。我们将 定量70种免疫标志物（细胞因子、趋化因子、生长因子）和代谢物的浓度 (GC/LC-MS）。这个项目的一个新颖之处是，我们将开发，验证， 对复杂的纵向数据应用新的统计方法，改进和适应现代结构建模 和组成微生物组数据分析。这些方法将提供一个严格的框架来处理 数据缺失和混淆一直是许多微生物组研究的主要限制。终极公众 健康影响是该项目的发现有可能为新疗法的开发提供信息， 或治疗GSM以促进女性健康。我们的团队拥有生物统计学，流行病学，衰老， 妇科学、基因组学、微生物学和免疫学，这些都是GSM系统生物学方法所必需的。

Over 50% of postmenopausal women are affected by the genitourinary syndrome of menopause (GSM), a progressive, chronic condition related to estrogen decline that includes vaginal atrophy, vaginal dryness, reduced sexual desire, and other signs and symptoms. GSM symptoms are a costly public health concern and worsen if untreated. Estrogen decline in menopause is thought to lead to reduced accumulation of vaginal glycogen and shifts in vaginal microbiota. These include low vaginal Lactobacillus spp. levels in 50-80% of postmenopausal women. Lactobacillus spp. protect the urogenital tract from pathogens in part by lactic acid production and hypothesized anti-inflammatory properties, but whether lactobacilli play a functional role in GSM is unknown. Available treatments for vulvovaginal GSM symptoms have limitations. Some women are contraindicated for hormonal therapy or are concerned about side effects. While vaginal lubricants provide some relief, many are toxic to the vaginal epithelium, reduce lactobacilli, and raise urogenital infection risk. Thus, new ways to treat and prevent GSM are needed. Because lactobacilli are often reduced in menopause, vaginal microbiota are plausible treatment targets; however, probiotics alone have not been proven effective. Also, age-related changes in vaginal microbiota co-vary with metabolite and immune profiles; but, the cross- talk between these molecular features and their role in GSM is unknown. We hypothesize that core vaginal micro-environment biomarkers (VMB; e.g., microbial, metabolite, and immune profiles) reflect vaginal biological aging (V-BA) that increases in menopause but may be modifiable by lifestyle factors. Prospective studies are needed to identify core age-related VMB and determine how they affect GSM. To achieve this goal, we will leverage 2,301 archived cervicovaginal samples collected from 812 women aged 35-60 years with clinical visits every six months for two years (R01-CA123467). Specific Aims are to 1) quantify V-BA using VMB; 2) evaluate the longitudinal relationship between V-BA and GSM; 3) longitudinally assess the relationship of lifestyle factors on V-BA and VMB; 4) quantify longitudinal mediation by V-BA and VMB between lifestyle and GSM. Microbiota are already profiled by 16S rRNA gene amplicon sequencing (R21-AI107224). We will quantify concentrations of 70 immune markers (cytokines, chemokines, growth factors) and metabolites (GC/LC-MS) in cervicovaginal samples. A novelty of this proposed project is that we will develop, validate, and apply new statistical methods for complex longitudinal data that refine and adapt modern structural modeling and compositional microbiome data analyses. These methods will provide a rigorous framework to handle missing data and confounding that has been a major limitation in many microbiome studies. The ultimate public health impact is that findings from this project have potential to inform development of new therapy to prevent or treat GSM to promote women’s health. Our team has the expertise in biostatistics, epidemiology, aging, gynecology, genomics, microbiology, and immunology required for this systems-biology approach to GSM.