Mechanisms of PGRMC2 action in female reproduction

PGRMC2 在女性生殖中的作用机制

基本信息

批准号：
8701667
负责人：
James K Pru
金额：
$ 16.63万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8701667
关键词：
Accounting Agonist Apoptosis Apoptotic Attenuated Binding Proteins Biological Assay Biology Birth Breeding Cell Differentiation process Cell Line Cell Proliferation Cell Survival Cells Cholesterol Data Decidual Cell Reactions Development Disease Embryo Endometrium Epithelial Cell Proliferation Estradiol Estrogens Event Expressed Sequence Tags Family Female Fertility Gene Expression Profile Generations Genes Genetic Transcription Grant Granulosa-Lutein Cells Histology Hormones Human Hyperplasia In Vitro Infertility Knockout Mice Lead Ligands Luteal Cells Malignant Female Reproductive System Neoplasm Maps Mediating Mediator of activation protein Membrane Mitosis Modeling Mus Mutagenesis Mutant Strains Mice Nuclear Oocytes Outcome Ovarian Ovarian Cysts Ovarian Follicle Ovarian hormone Ovary Ovulation Partner in relationship Pathway interactions Phenotype Physiology Play Pregnancy Pregnancy Maintenance Premature Ovarian Failure Preparation Primates Progesterone Progesterone Receptors Progestins RU-5020 Reproduction Reproductive Physiology Resistance Rodent Role Signal Pathway Site Spottings Steroid biosynthesis Stress Stromal Cells Syndrome Technology Testing Transgenic Mice Universities Uterus Washington Woman activating transcription factor analog autocrine base clinically relevant endometriosis female reproductive system granulosa cell implantation in vivo mouse genome mullerian-inhibiting hormone mutant natural Blastocyst Implantation premature public health relevance receptor recombinase reproductive reproductive axis reproductive function response senescence transcriptome sequencing validation studies

项目摘要

DESCRIPTION (provided by applicant): Progesterone (P4) is an essential hormone that elicits its actions at each level of the reproductive axis. In the uterus, P4 attenuates estradiol-induced epithelial cell proliferation and facilitates cellular differentiation in preparation for the establishment and maintenance of pregnancy. In the ovary, P4 acts directly on granulosa cells to inhibit mitosis and apoptosis despite these cells lacking expression of the classical progesterone receptor (PGR; i.e., PRA and PRB). P4 also promotes the viability and steroidogenic potential of luteal cells and stimulates both its own secretion and cholesterol synthesis. Many of the actions of P4 within the ovary and uterus are mediated by the PGR. However, not all of the actions of P4 can be explained by activation of PGR, since a number of cell lines that do not express this receptors, as well as Pgr null mice, are able to respond to P4. Our recent in vitro studies of ovarian and uterine cells have revealed that some actions of P4 such as granulosa/luteal cell viability, P4 synthesis and uterine stromal cell differentiation are mediated in part through the P4 binding protein, Progesterone Receptor Membrane Component-1 (PGRMC1). Through the use of conditional mutagenesis, we have established that Pgrmc1 is essential for normal fertility in that female mice lacking PGRMC1 display a subfertility phenotype and undergo premature ovarian failure (POF). This phenotype is also observed in some women where lower Pgrmc1 expression and haploinsufficiency associate with POF and polycystic ovarian syndrome. Pgrmc1 conditional KO (cKO) females also have uterine hyperplasia and develop both uterine and ovarian cysts. There is mounting evidence that PGRMC2 also plays an important role in female reproduction in that decreased PGRMC2 associates with advanced endometriosis in a primate model and may serve as a marker for the onset of parturition. These cumulative studies along with numerous in vitro studies clearly demonstrate that PGRMC1 and PGRMC2 play important and clinically relevant roles in regulating uterine and ovarian functions and that alteration in their expression results in the manifestation of disease states in the female reproductive system. In this grant we will now test our hypothesis that Pgrmc2 plays a fundamental role in uterine and ovarian physiology through the use of Pgrmc2 cKO mice. We also propose to assess fertility in female mice that are deficient in both Pgrmc1 and Pgrmc2 (double cKO mice). Breeding trials will initially be completed to determine fertility status of cKO mice. Other parameters to be evaluated in Aim 1 include uterine decidualization, ovulation, ovarian steroidogenesis, oocyte quality, and embryo implantation. In Aim 2, we will evaluate the importance of Pgrmc2 in mediating the anti-proliferative and anti-apoptotic actions of P4 in the ovary, as well as in the estrogen stimulated uterus. Finally, it is clear from studies of PGR mutant mice that PGR is not the sole mediator of P4-induced gene transcription. In Aim 2, we plan to also evaluate P4-induced transcriptional responses in the uterus and ovary of Pgrmc1/2 double cKO mice using RNA-seq. The successful completion of the proposed studies will provide compelling evident to support a role for PGRMC2 in female reproductive physiology. Establishing PGRMC2 as a mediator of specific uterine and ovarian functions will allow for the development of a new and selective class of P4 antagonists/agonists that target the PGRMC family.

描述（由申请人提供）：孕酮（P4）是一种必需的激素，它在生殖轴的每个级别都引起其作用。在子宫中，P4减弱了雌二醇诱导的上皮细胞增殖，并促进细胞分化，以制备建立和维持妊娠。在卵巢中，尽管这些细胞缺乏经典的孕酮受体表达（PGR；即PRA和PRB），但P4直接作用于颗粒细胞以抑制有丝分裂和凋亡。 P4还促进了黄体细胞的生存力和类固醇生成潜力，并刺激其自身分泌和胆固醇的合成。卵巢和子宫内P4的许多作用都是由PGR介导的。但是，并非所有P4的动作都可以通过PGR的激活来解释，因为许多未表达该受体的细胞系以及PGR NULL小鼠都能够对P4做出反应。我们最近对卵巢和子宫细胞的体外研究表明，P4的某些作用，例如颗粒/黄体细胞生存力，P4合成和子宫基质细胞分化，部分通过P4结合蛋白，孕酮受体受体膜成分-1（PGRMC1）介导。通过使用条件诱变，我们已经确定PGRMC1对于正常生育至关重要，因为缺乏PGRMC1的雌性小鼠表现出副作用表型并发生早产卵巢衰竭（POF）。在某些女性中也观察到这种表型，其中较低的PGRMC1表达和与POF和多囊卵巢综合征相关的单倍体不足。 PGRMC1有条件的KO（CKO）雌性还具有子宫增生，并同时发展子宫和卵巢囊肿。有越来越多的证据表明，PGRMC2在女性繁殖中也起着重要作用，因为在灵长类动物模型中，PGRMC2与晚期子宫内膜异位症相关，并且可以作为分娩开始的标志。这些累积研究以及众多的体外研究清楚地表明，PGRMC1和PGRMC2在调节子宫和卵巢功能中起重要且与临床相关的作用，并且其表达的改变导致女性生殖系统中疾病状态的表现。在这笔赠款中，我们将通过使用PGRMC2 CKO小鼠在子宫和卵巢生理学中发挥基本作用，即PGRMC2在子宫和卵巢生理中起着基本作用。我们还建议评估PGRMC1和PGRMC2（双CKO小鼠）不足的雌性小鼠的生育能力。最初将完成育种试验，以确定CKO小鼠的生育状况。在AIM 1中进行评估的其他参数包括子宫骨化，排卵，卵巢类固醇生成，卵母细胞质量和胚胎植入。在AIM 2中，我们将评估PGRMC2在介导卵巢中P4的抗增殖和抗凋亡作用方面的重要性，以及在雌激素刺激的子宫中的重要性。最后，从对PGR突变小鼠的研究可以清楚地看出，PGR不是P4诱导的基因转录的唯一介体。在AIM 2中，我们计划使用RNA-Seq评估PGRMC1/2双CKO小鼠的子宫和卵巢中P4诱导的转录反应。拟议的研究的成功完成将为支持PGRMC2在女性生殖生理学中的作用提供明显的说服力。建立PGRMC2作为特定子宫和卵巢功能的调解人将允许开发针对PGRMC家族的新型P4拮抗剂/激动剂。