Role of mineralocorticoid receptor in primate ovarian function

盐皮质激素受体在灵长类动物卵巢功能中的作用

• 批准号: 7471142
• 负责人: CHARLES L CHAFFIN
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7471142
• 关键词: Aging; Agonist; American; Animal Model; Anovulation; Attenuated; Body of uterus; Bolus Infusion; CYP21A2 gene; Cell Nucleus; Centers for Disease Control and Prevention (U.S.); Contraceptive Agents; Contraceptive methods; Corpus Luteum Maintenance; Couples; Cytoplasm; Data; Deoxycorticosterone; Derivation procedure; Detection; Development; Estradiol; Feedback; Gene Targeting; Granulosa-Lutein Cells; Health; Human; Human Chorionic Gonadotropin; In Vitro; Infertility; Intervention; Knowledge; Laboratories; Link; Longevity; Luteal Phase; Luteinization; Luteolysis; Macaca; Macaca fascicularis; Macaca mulatta; Maintenance; Malignant neoplasm of ovary; Menopause; Menstrual cycle; Messenger RNA; Methods; Mineralocorticoid Receptor; Mineralocorticoids; Modeling; Morbidity - disease rate; Numbers; Oral; Outcome; Ovarian; Ovarian Hyperstimulation Syndrome; Ovarian Stimulations; Ovary; Pathology; Pathway interactions; Periodicity; Physiological; Physiology; Pilot Projects; Plague; Pregnancy loss; Primates; Process; Production; Progesterone; Proteins; Public Health; Reproduction; Research; Role; Signal Transduction; Source; Steroid 21-Monooxygenase; Steroids; Stimulus; System; Testing; Time; United States; Vision; Woman; Work; clinically significant; corpus luteum; granulosa cell; in vivo; mortality; nonhuman primate; novel; prevent; receptor; research study; species difference; stem; therapy development

DESCRIPTION (provided by applicant): The formation, function, and demise of the primate corpus luteum are critical aspects of menstrual cyclicity, infertility, and contraceptive intervention. Recent work by the applicant's laboratory has demonstrated the presence of an intra-ovarian mineralocorticoid system in the non-human primate. In rhesus monkeys undergoing controlled ovarian stimulation, an ovulatory stimulus increases the expression of 21-hydroxylase (CYP21A2) mRNA in a time-dependent manner. CYP21A2 converts progesterone to the potent mineralocorticoid receptor (MR) agonist 11-deoxycorticosterone (DOC), and thus correlates with the observation that the ovulatory hCG bolus increases intra-follicular levels of DOC. In order to verify an ovarian origin for DOC, granulosa cells were isolated prior to an ovulatory stimulus and cultured in the presence of hCG, resulting in a dramatic increase in media concentrations of DOC. MR is expressed in the cytoplasm of both luteinizing granulosa cells as well as the corpus luteum until the mid to mid-late luteal phase, at which time MR is translocated to the nucleus. Importantly, antagonism of MR during luteinization attenuates hCG-induced progesterone, indicating that cytoplasmic MR has a key role in luteinization of granulosa cells in vitro. This R21 application proposes to test the overall hypothesis that MR is critical during the lifespan of the corpus luteum with two specific aims using the non-human primate model. Specific aim 1 will determine if MR has a critical role in the development of the corpus luteum using an MR antagonist. It is expected that antagonism of MR in vivo will prevent the critical rise in progesterone, leading to anovulation and incomplete luteal formation. Specific aim 2 will test the hypothesis that MR promotes luteal function through progesterone synthesis, and that intra-luteal levels of DOC increase in parallel with progesterone. In both aims, specific gene targets of MR will be elucidated. These studies will be the first to demonstrate a key role for MR in the ovarian physiology of any species, and are expected to further our understanding of corpus luteum development, maintenance, and demise in a biomedically relevant primate model. PUBLIC HEALTH RELEVANCE: Pathologies associated with the ovary carry a high morbidity and mortality, largely through difficulty in detection, but also from a lack of treatment stemming from a rather incomplete understanding of basic ovarian processes. These include, for example, PCOS, ovarian cancer, and ovarian hyperstimulation syndrome (OHSS). In addition, infertility plagues a growing number of couples in the United States (>6 million; Center for Disease Control); a proportion of this infertility can be linked to insufficiency of P production during the luteal phase, although the underlying causes of this are completely unknown. Novel contraception remains a problem, with a large preponderance of North American women continuing to rely on derivations of combined oral contraception and no new methods in sight. Aging and reproduction is clearly a growing topic in the United States, and a better knowledge of basic ovarian processes will facilitate understanding the menopausal transition in women. These issues all support the idea that ovarian physiology is a relevant topic with profound health outcomes. Overall, these exploratory studies should reveal a novel signaling steroid pathway in the primate ovary that could have clinical significance for contraceptive development, infertility, and potentially ovarian cancer.

描述（由申请人提供）：灵长类黄体的形成、功能和消亡是月经周期、不孕和避孕干预的关键方面。申请人实验室最近的工作证明了非人类灵长类动物卵巢内盐皮质激素系统的存在。在接受受控卵巢刺激的恒河猴中，排卵刺激会以时间依赖性方式增加 21-羟化酶 (CYP21A2) mRNA 的表达。 CYP21A2 将黄体酮转化为强效盐皮质激素受体 (MR) 激动剂 11-脱氧皮质酮 (DOC)，因此与排卵 hCG 推注增加卵泡内 DOC 水平的观察结果相关。为了验证 DOC 的卵巢来源，在排卵刺激之前分离颗粒细胞，并在 hCG 存在的情况下培养，导致 DOC 培养基浓度急剧增加。 MR 在黄体化颗粒细胞和黄体的细胞质中表达，直至黄体期中后期，此时 MR 易位至细胞核。重要的是，黄体化过程中 MR 的拮抗作用减弱了 hCG 诱导的黄体酮，表明细胞质 MR 在体外颗粒细胞黄体化中具有关键作用。该 R21 应用建议使用非人类灵长类动物模型测试 MR 在黄体生命周期中至关重要的总体假设，具有两个特定目标。具体目标 1 将使用 MR 拮抗剂确定 MR 是否在黄体发育中发挥关键作用。预计体内 MR 的拮抗作用将阻止黄体酮的急剧升高，从而导致无排卵和黄体形成不完全。具体目标 2 将检验以下假设：MR 通过黄体酮合成促进黄体功能，并且黄体内 DOC 水平与黄体酮平行增加。在这两个目标中，MR 的特定基因靶点都将得到阐明。这些研究将首次证明 MR 在任何物种的卵巢生理学中的关键作用，并有望进一步加深我们对生物医学相关灵长类动物模型中黄体发育、维持和死亡的理解。公共健康相关性：与卵巢相关的病理具有很高的发病率和死亡率，这主要是由于难以检测，但也由于对卵巢基本过程的不完全了解而导致缺乏治疗。这些包括，例如，多囊卵巢综合症、卵巢癌和卵巢过度刺激综合征（OHSS）。此外，不孕不育困扰着美国越来越多的夫妇（超过 600 万；疾病控制中心）；这种不孕症的一部分可能与黄体期磷生成不足有关，尽管其根本原因尚不清楚。新型避孕方法仍然是一个问题，大部分北美女性仍然依赖复方口服避孕药的衍生品，而且看不到新的方法。在美国，衰老和生殖显然是一个日益增长的话题，更好地了解卵巢的基本过程将有助于了解女性的更年期过渡。这些问题都支持这样的观点：卵巢生理学是一个具有深远健康影响的相关主题。总体而言，这些探索性研究应该揭示灵长类动物卵巢中的一种新的信号传导途径，该途径可能对避孕药开发、不孕症和潜在的卵巢癌具有临床意义。