Anti-Mullerian hormone actions to control primate folliculogenesis

 描述(申请人提供)：本研究的目的是了解在月经周期中，抗苗勒氏激素(AMH)直接或间接通过其他局部合成因子以雄鹿依赖的方式调节灵长类动物腔前卵泡生长和腔卵泡成熟的机制。越来越多的证据表明，AMH改变了腔前卵泡的生长；AMH减少培养的颗粒细胞芳香化酶和黄体生成素受体的表达，这为AMH在腔卵泡中可能的作用增加了另一个维度。同样，PI的证据表明，AMH在猕猴卵泡中异质性和动态表达，这加强了AMH在灵长类卵泡发生中的阶段依赖性作用的概念。最后，最近的“体外卵泡成熟”(IFM)技术为揭示灵长类卵巢中对卵泡发育和卵母细胞成熟至关重要的局部因子和信号通路提供了一个可行的模型。因此，我们建议在成年雌性恒河猴身上进行翻译研究，以验证以下假设：(1)AMH促进腔前卵泡的生长，但在体外抑制腔前卵泡的成熟；(2)AMH促进腔前到腔卵泡的生长，但在体内阻止优势卵泡的进一步成熟和选择。AMH添加(重组人AMH蛋白)和消融(抗人AMH抗体)实验将通过体外IFM和体内卵巢内输注进行。将在辅助生殖技术支持核心和大规模并行测序共享资源的帮助下，分析卵泡的形态和组织学，以及功能I类固醇的发生和局部因子的产生，卵母细胞的成熟和受精和着床前发育的能力，以及全基因组mRNA水平和选定的AMH调节基因(AIM 3)。在IFM期间，AMH也将被检测为进一步的卵泡生长和卵母细胞成熟的非侵入性生物标志物。最后，非人类灵长类动物研究产生的数据将应用于人类IFM，假设AMH操作改善了人类卵泡及其封闭的卵母细胞的协调发育和成熟(目标4)。这些实验将为研究AMH在灵长类动物卵泡中的异质性表达和阶段依赖性作用，以及AMH控制卵泡发育的机制提供有价值的信息。在IFM期间，AMH作为卵泡生长和卵母细胞能力的生物标志物的潜力将出现新的见解。新出现的概念应该完善临床范例，例如，局部AMH作为卵巢储备的生物标志物在不孕症中的作用或价值，包括原发卵巢功能不全、多囊卵巢综合征、子宫内膜异位症、体外受精、生殖衰老和癌症。

 DESCRIPTION (provided by applicant): The goal of this research is to understand the mechanisms whereby anti-Müllerian hormone (AMH) either directly, or indirectly via other locally synthesized factors, regulates preantral follicle growth and antral follicle maturation in the stag-dependent manner in primates during the menstrual cycle. Increasing evidence suggests that AMH alters preantral follicle growth; and the discovery that AMH reduces aromatase and luteinizing hormone receptor expression in cultured granulosa cells adds another dimension to possible AMH actions in antral follicles. Likewise, the PI's evidence that AMH is heterogeneously and dynamically expressed in the macaque follicles reinforces the concept of stage-dependent AMH actions during primate folliculogeneisis. Finally, the recent "in vitro follicl maturation" (IFM) technique provides a feasible model for manipulative studies to unravel local factors and signaling pathways that are critical to the follicular development and oocyte maturation in the primate ovary. Therefore, translational studies are proposed in the adult, female rhesus monkeys, to test the hypotheses that: (Aim 1) AMH promotes preantral follicle growth, but inhibits antral follicle maturation in vitro and (Aim 2) AMH promotes preantral-to-antral follicle growth, but prevents further maturation and selection of the dominant follicle durig the follicular phase of the menstrual cycle in vivo. AMH addition (recombinant human AMH protein) and ablation (anti-human AMH antibody) experiments will be conducted through IFM in vitro and intraovarian infusion in vivo. Follicular morphology and histology, as well as function i steroidogenesis and local factor production, oocyte maturation and competence in fertilization and preimplantation development, plus genome-wide mRNA levels and selected AMH-regulated genes (Aim 3) will be analyzed with the assistance of the Assisted Reproductive Technologies Support Core and Massively Parallel Sequencing Shared Resource. AMH will also be examined as a noninvasive biomarker for further follicle growth and oocyte maturation during IFM. Finally, data generated from the nonhuman primate research will be applied to human IFM with the hypothesis that AMH manipulation improves the coordinated development and maturation of human follicles and their enclosed oocytes (Aim 4). These experiments will provide valuable information on the heterogeneous expression and stage- dependent actions of AMH in ovarian follicles, as well as mechanisms of AMH controlling follicular development, in primates. New insight will emerge on the potential of AMH to serve as a biomarker for follicle growth and oocyte competence during IFM. Emerging concepts should refine clinical paradigms, e.g., local AMH action or value as a biomarker of ovarian reserve in infertility, including primary ovarian insufficiency, polycystic ovarian syndrome, endometriosis, in vitro fertilization, reproductive aging and cancer.