Vaginal microbiota, immune responses and vulvovaginal symptoms during menopause

描述(申请人提供)：研究表明，25%-50%的更年期妇女报告外阴阴道症状，包括阴道干燥、刺激、灼热、瘙痒和性交困难，所有这些都严重影响生活质量。随着女性接近更年期，雌激素水平下降伴随着阴道的变化，包括上皮糖原积累减少，pH值升高，阴道组织恶化，导致阴道不适症状。激素治疗是绝经妇女外阴阴道症状的主要治疗方法，但在一些妇女中是禁忌。改善绝经后妇女症状和生活质量的非激素治疗方法尚未得到很好的研究。阴道微生物群落(称为阴道微生物区系)在保护女性生殖道免受疾病侵袭方面发挥了关键作用；然而，令人惊讶的是，人们对绝经过渡不同阶段阴道细菌的组成、个体之间的差异、与阴道免疫微环境的相关性、与妇科疾病和性功能的相关性知之甚少。在2009-2012年间，885名年龄在35-60岁之间的女性参加了一项关于更年期转变的前瞻性研究，并在两年内每六个月跟踪一次(R01-CA123467)。利用这一资源，这项拟议的研究将利用储存在冷冻库中的2500份纵向阴道样本来解决以下三个具体目标：(1)描述两年内按年龄和绝经阶段的阴道微生物区系的变化，(2)评估阴道微生物区系与外阴阴道不适之间的关系，以及(3)评估阴道免疫微环境与微生物区系之间的相关性。根据生殖老龄化研讨会(STRAW)指南，参与者根据更年期类别进行分类。从临床医生收集的阴道中部拭子中分离出的完整基因组DNA中扩增出的16S rRNA基因分析将对阴道微生物区系进行表征。宫颈分泌物将被用来量化27种不同的免疫标志物(细胞因子、趋化因子和生长因子)的浓度，使用基于总蛋白调整的多重珠状免疫分析。通过汇集生物信息学、流行病学、基因组学、妇科学、微生物学和统计学的多学科专业知识，这项研究将能够提供关于(1)绝经阶段阴道微生物组的组成和稳定性的差异，(2)微生物变化与外阴阴道不适之间的关联，以及(3)绝经过渡期间外阴阴道微生物和免疫微环境变化之间的可能相关性的关键描述性数据。这些数据将用于评估非激素、抗炎或益生菌干预治疗围绝经期和绝经后妇女常见外阴阴道症状的可能性。

DESCRIPTION (provided by applicant): Studies demonstrate that between 25-50% of menopausal women report vulvovaginal symptoms including vaginal dryness, irritation, burning, itching, and dyspareunia, all of which severely affect quality of life. As women approach menopause, decreased estrogen levels are accompanied by changes in the vagina including decreased glycogen accumulation in the epithelium, increased pH, and deterioration of vaginal tissues, which lead to symptoms of vaginal discomfort. Hormone therapy is the primary treatment for vulvovaginal symptoms in menopausal women, however it is contraindicated in some women. Non-hormonal therapeutic approaches for improving symptoms and quality of life in postmenopausal women have not been well studied. The vaginal microbial communities (termed the vaginal microbiota) play a critical role in protecting the female genital tract from disease; however, surprisingly little is known about the composition of vaginal bacteria across the different stages of menopausal transition, how they differ between individuals, the correlation with vaginal immunologic microenvironment, or the associations with gynecologic complaints and sexual functioning. Between 2009-2012, 885 women aged 35-60 years were enrolled in a prospective study of menopause transition and followed every six months for two years (R01-CA123467). Taking advantage of that resource, the proposed study will utilize 2500 longitudinal vaginal samples stored in a freezer archive to address the following three specific aims: (1) To describe the changes in the vaginal microbiota by age and stage of menopause over a 2-year period, (2) To evaluate the association between the vaginal microbiota and vulvovaginal complaints and (3) To assess the correlation between the vaginal immunologic microenvironment with the microbiota. Participants were categorized according to menopausal categories by the Stages of Reproductive Aging Workshop (STRAW) guidelines. The vaginal microbiota will be characterized using 16S rRNA gene analysis amplified from whole genomic DNA isolated from clinician-collected mid-vaginal swabs. Cervical secretions will be used to quantify concentrations of 27 different immune markers (cytokines, chemokines, and growth factors) using multiplexed bead-based immunoassays with total protein adjustment. By bringing together multidisciplinary expertise in bioinformatics, epidemiology, genomics, gynecology, microbiology and statistics, this study will be able to provide critical descriptive data on (1) differences in the composition and stability of the vaginal microbiome by menopausal stage, (2) association between microbial changes and presence of vulvovaginal complaints, and (3) the possible correlation between vulvovaginal microbial and immunologic microenvironmental changes during the menopausal transition. These data will be used to evaluate the potential for non-hormonal, anti-inflammatory, or probiotic interventions for the treatment of common vulvovaginal complaints in peri- and postmenopausal women.