Uterine microbiome: Functional significance in establishing the uterine program postpartum

项目总结 大约三分之一的产后奶牛在产后一周内患上子宫疾病(子宫炎)。 5%的产犊会导致奶牛在30天内因子宫炎或其他并发症而死亡 起源于产犊期间。产后子宫感染在女性中较少见，但约10%的 在美国，与怀孕有关的死亡是由产后生殖道感染引起的。早起的奶牛 产后子宫疾病会发展为慢性子宫炎症(子宫内膜炎或亚临床 子宫内膜炎；姐妹染色单体交换)。女性的慢性子宫内膜炎(盆腔炎性疾病)类似于牛 该病由多种细菌引起。如果不进行治疗，姐妹染色单体交换和前列腺癌都会导致不孕。 无法有效治疗子宫疾病和恢复生育能力的部分原因可能是关键的差距 了解子宫微生物群及其与疾病状态、子宫免疫功能、 炎症、组织再生和子宫功能障碍的长期规划。这样做的目的是 建议了解子宫疾病的起源以及子宫疾病通过 计划不育和不育。研究始于处女牛子宫，在那里存在和 将确定第一次繁殖后微生物组的生存能力(目标1；研究1A和1B)。黄曲霉的微生物群 然后将研究第一次和第二次怀孕(目标1，研究1C)。早期是否有子宫炎 产后期通过结构和功能机制永久性损害子宫内膜 全身性抗生素治疗是否减轻这种影响是目标2的主题。最后，胚胎营养 将评估患病子宫内膜的容量(目标3)。总的来说，拟议的调查将 阐明微生物组计划生育的新的结构和功能机制。这个 无论是从概念上还是从技术角度来看，研究都是创新的。子宫微生物群及其作用 生育率是一个令人振奋的新研究领域，在概念上也是创新的。奶牛提供了一种创新的 子宫疾病研究的模型，因为子宫疾病状态和 奶牛和妇女的结果。全基因组元基因组测序将作为这种方法来执行 将使我们能够评估整个微生物群。转录组将用激光捕获进行研究。 显微切割(LCM)和RNA测序。减少代表性的亚硫酸盐测序(RRBS)将是 用于识别与转录组变化相关的DNA甲基化模式。建议数 研究将确定子宫内膜不能建立接受性的组织特异性机制 子宫环境和奶牛不能怀孕。这一基础知识对于 开发诊断、治疗和改善子宫疾病影响的创新工具。如果疾病 了解了机制后，就有可能防止组织损伤和(或)逆转 制定方案，全面恢复患有子宫病的妇女和/或牛的生育能力。

PROJECT SUMMARY Approximately one-third of postpartum cows develop uterine disease (metritis) within one week after calving and 5% of all calvings result in death of the cow within 30 days due to metritis or other complications that originate during calving. Postpartum uterine infection in women is less common, but approximately 10% of pregnancy-related deaths in the United States are caused by puerperal genital tract infection. Cows with early postpartum uterine disease will develop chronic inflammation of the uterus (endometritis or subclinical endometritis; SCE). Chronic endometritis in women (pelvic inflammatory disease; PID) is similar to the bovine disease and is caused by a variety of bacterial species. If left untreated, both SCE and PID cause infertility. The inability to effectively treat uterine disease and restore fertility may be partially explained by a critical gap in understanding of the uterine microbiome and its relationship to the disease status, uterine immune function, inflammation, tissue regeneration, and long-term programming of uterine dysfunction. The goal of this proposal is to understand the origins of uterine disease and the mechanisms through which uterine disease programs subfertility and infertility. The studies begin with the virgin bovine uterus where the presence and viability of the microbiome after first breeding will be determined (Aim 1; Study 1A & 1B). The microbiome of first and second pregnancy will then be studied (Aim 1, Study 1C). Whether metritis during the early postpartum period permanently damages the endometrium through structural and functional mechanisms and whether systemic antibiotic treatment mitigates the effect is the subject of Aim 2. Finally, the embryotrophic capacity of the diseased endometrium will be assessed (Aim 3). Collectively, the proposed investigations will elucidate novel structural and functional mechanisms through which the microbiome programs fertility. The research is innovative from both a conceptual and technical perspective. The uterine microbiome and its effect on fertility is an exciting new area of research and conceptually innovative. The cow provides an innovative model for uterine disease research due to the remarkable similarity between uterine disease states and outcomes in cows and women. Whole genome metagenomic sequencing will be performed as this approach will enable us to assess the entire microbiome. The transcriptome will be studied by using laser capture microdissection (LCM) and RNA sequencing. Reduced representation bisulfite sequencing (RRBS) will be used to identify patterns of DNA methylation associated with changes in the transcriptome. The proposed studies will identify tissue-specific mechanisms through which the endometrium fails to establish a receptive uterine environment and the cow fails to become pregnant. This fundamental knowledge is necessary to develop innovative tools to diagnose, treat, and ameliorate the effects of uterine disease. If disease mechanisms are understood then it may be possible to prevent tissue damage and (or) reverse the programming to fully restore fertility in women and or cattle that experience uterine disease.