Anti-Mullerian hormone actions to control primate folliculogenesis

 描述（由申请人提供）：本研究的目的是了解抗苗勒管激素（AMH）直接或通过其他局部合成因子间接调节灵长类动物月经周期中窦前卵泡生长和窦卵泡成熟的阶段依赖性机制。越来越多的证据表明，AMH改变腔前卵泡的生长; AMH降低培养颗粒细胞中芳香化酶和黄体生成素受体表达的发现为AMH在有腔卵泡中的可能作用增加了另一个维度。同样，PI的证据表明，AMH是异质性和动态表达的猕猴卵泡加强了灵长类动物卵泡发育过程中的阶段依赖性AMH的行动的概念。最后，最近的“体外卵泡成熟”（IFM）技术提供了一个可行的模型，操纵研究，以解开当地的因素和信号通路，是至关重要的卵泡发育和卵母细胞成熟的灵长类动物卵巢。因此，我们提出了在成年雌性恒河猴中进行转化研究，以验证以下假设：（目的1）AMH促进腔前卵泡生长，但抑制体外腔卵泡成熟;（目的2）AMH促进腔前至腔卵泡生长，但阻止月经周期卵泡期优势卵泡的进一步成熟和选择。将通过体外IFM和体内卵巢内输注进行AMH添加（重组人AMH蛋白）和消融（抗人AMH抗体）实验。卵泡形态学和组织学，以及类固醇生成和局部因子产生的功能，卵母细胞成熟和受精和植入前发育的能力，加上全基因组mRNA水平和选定的AMH调节基因（目标3）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进行分析。AMH也将作为IFM期间进一步卵泡生长和卵母细胞成熟的非侵入性生物标志物进行检查。最后，从非人灵长类动物研究中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卵母细胞的协调发育和成熟（目标4）。 这些实验将为AMH在灵长类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。AMH作为IFM期间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。新兴的概念应该完善临床范例，例如，局部AMH作为不孕症卵巢储备生物标志物的作用或价值，包括原发性卵巢功能不全、多囊卵巢综合征、子宫内膜异位症、体外受精、生殖老化和癌症。

 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.