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）：黄体酮 (P4) 是一种重要激素，可在生殖轴的各个层面发挥作用。在子宫中，P4 可减弱雌二醇诱导的上皮细胞增殖并促进细胞分化，为妊娠的建立和维持做好准备。在卵巢中，P4 直接作用于颗粒细胞，抑制有丝分裂和细胞凋亡，尽管这些细胞缺乏经典孕激素受体（PGR；即 PRA 和 PRB）的表达。 P4 还可以促进黄体细胞的活力和类固醇生成潜力，并刺激其自身分泌和胆固醇合成。 P4 在卵巢和子宫内的许多作用都是由 PGR 介导的。然而，并不是 P4 的所有作用都可以通过 PGR 的激活来解释，因为许多不表达这种受体的细胞系以及 Pgr 无效小鼠能够对 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 的一些作用，例如颗粒/黄体细胞活力、P4 合成和子宫基质细胞分化，部分是通过 P4 结合蛋白、孕酮受体膜成分 1 (PGRMC1) 介导的）。通过使用条件诱变，我们已经确定 Pgrmc1 对于正常生育能力至关重要，因为缺乏 PGRMC1 的雌性小鼠会表现出生育力低下的表型并发生卵巢早衰 (POF)。这种表型也在一些女性中观察到，其中 Pgrmc1 表达较低和单倍体不足与 POF 和多囊卵巢综合征相关。 Pgrmc1 条件性 KO (cKO) 女性也会出现子宫增生，并同时出现子宫和卵巢囊肿。越来越多的证据表明 PGRMC2 在雌性生殖中也发挥着重要作用，因为 PGRMC2 的减少与灵长类动物模型中的晚期子宫内膜异位症相关，并且可能作为分娩开始的标志。这些累积研究以及大量体外研究清楚地表明，PGRMC1 和 PGRMC2 在调节子宫和卵巢功能中发挥着重要的临床相关作用，并且其表达的改变导致女性生殖系统疾病状态的表现。在这笔资助中，我们现在将通过使用 Pgrmc2 cKO 小鼠来检验我们的假设，即 Pgrmc2 在子宫和卵巢生理学中发挥重要作用。我们还建议评估缺乏 Pgrmc1 和 Pgrmc2 的雌性小鼠（双 cKO 小鼠）的生育能力。繁殖试验将首先完成以确定 cKO 小鼠的生育状态。目标 1 中要评估的其他参数包括子宫蜕膜化、排卵、卵巢类固醇生成、卵母细胞质量和胚胎植入。在目标 2 中，我们将评估 Pgrmc2 在介导 P4 在卵巢以及雌激素刺激的子宫中的抗增殖和抗凋亡作用中的重要性。最后，从 PGR 突变小鼠的研究中可以清楚地看出，PGR 并不是 P4 诱导的基因转录的唯一介质。在目标 2 中，我们还计划使用 RNA-seq 评估 Pgrmc1/2 双 cKO 小鼠子宫和卵巢中 P4 诱导的转录反应。拟议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令人信服的证据。将 PGRMC2 确立为特定子宫和卵巢功能的调节剂将有助于开发针对 PGRMC 家族的新型选择性 P4 拮抗剂/激动剂。