The Vaginal Microbiome: Disease, Genetics and the Environment

描述：阴道是与环境的交互界面，因此被一层保护性的上皮表面覆盖。细菌和其他微生物通过各种机制进一步保护宿主免受病原体的入侵。正常阴道微生物群的变化，特别是与细菌性阴道病相关的微生物群的变化，被认为是导致中期自然妊娠丢失和自发早产的风险。此外，阴道微生物群的改变可能会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)。寄主条件的生理变化(例如，更年期和怀孕)正开始被研究为改变“正常”菌群的潜在选择性条件，其对疾病易感性和传播的影响仍有待更明确地阐明。慢性异常生理状态(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研究领域是，在正常和生理变化的情况下，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。这项拟议的研究将解决我们知识中的空白，并阐明阴道微生物群如何在不同人群中对产科不良结局和性传播疾病做出贡献。该项目的目标是回答以下问题：具体目标1.宿主的基因对阴道微生物组的组成有贡献吗？我们假设，女性的基因构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。这一目标分为两个分目标，第一个分目标将比较和量化大西洋中部双胞胎登记处(MATR)中同卵(MZ)和异卵(DZ)双胞胎阴道内的微生物种群。第二个分目标将利用从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之间是否存在关系的问题。具体目的2.阴道微生物群的哪些变化与常见的生理紊乱或宿主的非感染性病理状态有关？我们假设“改变”的生理(怀孕、更年期)和病理(慢性病、子宫切除)条件，或环境“暴露”(外源性激素、抗生素、慢性免疫抑制药、吸烟；冲洗)可以而且经常确实可预测地改变阴道微环境。这些变化反过来会导致阴道内微生物种群的变化。微生物种群的变化可能会对受影响个人的自发性和未来的福祉产生积极或更有可能消极的影响。我们将描述这些“改变”的生理和病理条件的影响，以及环境暴露对阴道微生物组组成的影响，并测试它们导致阴道微生物组可预测变化的假设。将评估微生物组的分子分析与基于Amsel标准的实验室结果和Nugent评分之间的关系。具体目标3.阴道微生物群的哪些变化与相关的传染病和条件有关？我们将检验这一假设，即传染病可预测地改变阴道微生物群，并且这些变化对疾病过程有影响。我们还将检验这样一个假设，即阴道微生物群对某些相关传染病的易感性有影响。在这一特定目标下进行的研究将提供一个平台，用于模拟生理、病理、环境和民族/种族因素的影响及其相互作用，以确定阴道微生物群。除了提供关键的描述性数据外，这将是一个产生假设的具体目标。我们将分析以下人群的阴道微生物群：正常育龄妇女；有常见病理疾病(如阴道病、阴道炎、病毒感染、细菌性性病)的同龄妇女；三种不同民族/种族的妇女：欧洲高加索人、非裔美国人和墨西哥西班牙人。我们将结合使用高通量的“下一代”测序技术来解决这些问题，包括罗氏454FLX和升级后的Illumina基因组分析仪II仪器，目前安装在VCU的核酸研究设施中。

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.