The Vaginal Microbiome: Disease, Genetics and the Environment

描述：阴道是一个与环境交互的界面，因此被保护性上皮表面覆盖。反过来，这个表面被细菌和其他微生物定植，这些微生物通过各种机制进一步保护宿主免受病原体的入侵。正常阴道菌群的改变，特别是与细菌性阴道病相关的改变，被认为会增加妊娠中期自然流产和自然早产的风险。此外，阴道微生物组的改变可能增加某些病原体传播的可能性，包括人类免疫缺陷病毒1型（HIV-1）。寄主状态的生理变化（例如，更年期和怀孕）已开始作为“正常”菌群变化的潜在选择性条件进行研究，它们对疾病易感性和传播的影响仍有待更明确地阐明。长期异常生理状态（如糖尿病）对正常阴道菌群的影响尚未得到很好的描述或研究。最后，一个几乎未被探索的调查领域是在正常和生理改变的情况下，包括种族/民族在内的遗传因素对建立和维持“正常”阴道菌群的作用。拟议的研究将解决我们知识上的差距，并阐明阴道微生物群如何在不同人群中导致不良的产科结果和性传播疾病。该项目的目标旨在回答以下问题：具体目标1。宿主的基因对阴道微生物群的组成有影响吗？我们假设女性的基因组成显著影响某些共生、寄生和致病微生物定植和/或感染生殖道的能力。这一目标分为两个子目标，第一个目标将比较和量化中大西洋双胞胎登记处（MATR）中单卵双胞胎（MZ）和异卵双胞胎（DZ）阴道内的微生物种群。第二个子目标将利用从DZ和MZ双胞胎收集的样本来解决阴道、口腔和胃肠道微生物群之间是否存在关系的问题。具体目标2。阴道微生物组的哪些变化与宿主常见的生理扰动或非感染性病理状态有关？我们假设生理（怀孕、更年期）和病理（慢性疾病、子宫切除术）条件的“改变”，或环境“暴露”（外源性激素、抗生素、慢性免疫抑制剂、吸烟、灌洗）可以并且经常可以预测地改变阴道微环境。这些变化反过来又会导致阴道内微生物种群的变化。微生物种群的变化可能对受影响个体的自发和未来的福祉产生积极或更可能是消极的影响。我们将描述这些“改变”的生理和病理条件以及环境暴露对阴道微生物组组成的影响，并验证它们导致阴道微生物组可预测变化的假设。微生物组的分子分析与基于Amsel标准和纽金特评分的实验室结果之间的关系将被评估。具体目标3。阴道微生物组的哪些变化与相关的传染病和病症有关？我们将测试传染病可预测地改变阴道微生物群的假设，这些变化对疾病过程有影响。我们还将检验阴道微生物组对某些相关传染病的易感性有影响的假设。在这个特定目标中进行的研究将为生理、病理、环境和民族/种族因素的影响及其相互作用的建模提供一个平台，以确定阴道微生物组。除了提供关键的描述性数据外，这将是一个产生假设的特定目标。我们将分析阴道微生物组：正常育龄妇女；患有常见病理状况（如阴道病、阴道炎、病毒感染、细菌性性传播疾病）的同龄妇女；三个不同民族/种族群体的女性：欧洲高加索人、非洲裔美国人和墨西哥裔西班牙人。我们将使用高通量“下一代”测序技术解决这些问题，包括罗氏454 FLX和升级的Illumina基因组分析仪II仪器，目前安装在VCU的核酸研究设施。

DESCRIPTION: The vagina is an interactive interface with the environment, and as such is covered by a protective epithelial surface. This surface, in turn, is colonized by bacteria and other microorganisms which, through a variety of mechanisms serve to further protect the host from invasion by pathogens. Alterations in the normal vaginal microflora, particularly those associated with bacterial vaginosis, are thought to contribute to risk of spontaneous pregnancy loss in the second trimester and spontaneous preterm birth. Additionally, alterations in the vaginal microbiome may increase the likelihood of transmission of certain agents including human immunodeficiency virus type 1 (HIV-1). There are physiologic alterations in host condition (e.g., menopause and pregnancy), which are beginning to be investigated as potential selective conditions for change in the "normal" flora, and their impact on disease susceptibility and transmission remains to be more definitively elucidated. The effects of chronically abnormal physiologic states (e.g., diabetes mellitus) on normal vaginal flora have not been well described or studied. Finally, an almost unexplored area of inquiry is the genetic contribution, including race/ethnicity, to the establishment and maintenance of a "normal" vaginal flora, under normal and physiologically altered circumstances. The proposed research will address gaps in our knowledge and shed light on how the vaginal microbiome contributes to adverse obstetrical outcomes and sexually transmitted disease in diverse populations. The aims of the project are intended to answer the following questions: Specific Aim 1. Do the genes of the host contribute to the composition of the vaginal microbiome? We hypothesize that a woman's genetic composition significantly affects the ability of certain commensal, parasitic and pathogenic microbes to colonize and/or infect the genital tract. This aim is divided into 2 subaims, the first of which will compare and quantify the microbial populations inhabiting the vaginas of monozygotic (MZ) and dizygotic (DZ) twins in the Mid Atlantic Twin Registry (MATR). The second subaim will address the question of whether there is a relationship between the microbiomes of the vagina, mouth and GI tract utilizing samples collected from the DZ and MZ twins. Specific Aim 2. What changes in the vaginal microbiome are associated with common physiological perturbations or non-infectious pathological states of the host? We hypothesize that "altered" physiologic (pregnancy, menopause) and pathologic (chronic disease, hysterectomy) conditions, or environmental "exposures" (exogenous hormones, antibiotics, chronic immunosuppressant, smoking; douching) can and often do predictably alter the vaginal microenvironment. These alterations in turn will lead to alterations in microbial populations within the vagina. Changes in the microbial populations may have impacts, positive or more likely negative, on the spontaneous and future well-being of the affected individual. We will characterize the effects of these "altered" physiologic and pathologic conditions, and environmental exposures exert on the composition of the vaginal microbiome, and test the hypothesis that they lead to predictable changes in the vaginal microbiome. The relationship between the molecular analysis of the microbiome and laboratory findings based on Amsel's criteria and the Nugent Score will be evaluated. Specific Aim 3. What changes in the vaginal microbiome are associated with relevant infectious diseases and conditions? We will test the hypothesis that infectious diseases predictably alter the vaginal microbiome, and that these changes have an impact on the disease process. We will also test the hypothesis that the vaginal microbiome has an impact on susceptibility to some relevant infectious diseases. The research to be conducted in this Specific Aim will provide a platform for modeling the impact of physiological, pathological, environmental and ethnic/racial factors, and their interactions, in determining the vaginal microbiome. In addition to providing critical descriptive data, this will be a hypothesis-generating Specific Aim. We will analyze the vaginal microbiomes of: normal women of reproductive age; women in the same age range with common pathological conditions (e.g., vaginosis, vaginitis, viral infections, bacterial STDs); women of three different ethnic/racial groups: European Caucasian, African-American, and Mexican Hispanic. We will address these questions using a combination of high throughput 'nextgen' sequencing technologies, including the Roche 454 FLX and the upgraded Illumina Genome Analyzer II instruments currently installed in the Nucleic Acids Research Facilities at VCU.