Uterine microbiome: Functional significance in establishing the uterine program postpartum

项目摘要 大约三分之一的产后奶牛在产犊后一周内发生子宫疾病（子宫炎 5%的产犊在30天内由于子宫炎或其他并发症导致奶牛死亡， 产犊期间产生的。妇女产后子宫感染不太常见，但大约10%的妇女 在美国，与妊娠有关的死亡是由产褥期生殖道感染引起的。奶牛早期 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）， 结肠炎; SCE）。女性慢性盆腔炎（盆腔炎; PID）类似于牛 它是由多种细菌引起的。如果不治疗，SCE和PID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一个关键的差距 了解子宫微生物组及其与疾病状态，子宫免疫功能， 炎症、组织再生和子宫功能障碍的长期规划。这个目标 建议是了解子宫疾病的起源和机制，通过子宫疾病 计划生育力低下和不孕症。这些研究开始于处女牛子宫， 将确定第一次繁殖后微生物组的活力（目标1;研究1A和1B）。的微生物组 然后将研究第一次和第二次妊娠（目标1，研究1C）。早期子宫炎是否 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能减轻这种影响是目标2的主题。最后，胚胎营养型 将评估患病子宫内膜的容量（目标3）。总体而言，拟议的调查将 阐明了微生物组编程生育的新结构和功能机制。的 研究从概念和技术角度都是创新的。子宫微生物组及其影响 是一个令人兴奋的新研究领域，在概念上具有创新性。奶牛提供了一种创新的 由于子宫疾病状态和子宫内膜异位症之间的显著相似性， 奶牛和妇女的结果。全基因组宏基因组测序将作为这种方法进行 能让我们评估整个微生物组将使用激光捕获来研究转录组 显微切割（LCM）和RNA测序。还原代表性亚硫酸氢盐测序（RRBS）将在 用于鉴定与转录组变化相关的DNA甲基化模式。拟议 研究将确定组织特异性机制，通过该机制，子宫内膜未能建立一个接受 子宫环境和母牛未能怀孕。这些基本知识对于 开发创新的工具来诊断，治疗和改善子宫疾病的影响。如果疾病 如果了解了这些机制，那么就有可能防止组织损伤和（或）逆转组织损伤。 制定方案，以完全恢复患有子宫疾病的妇女和/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.