PGRMC1 function in female reproductive physiology

PGRMC1 在女性生殖生理学中的功能

• 批准号: 7867760
• 负责人: JOHN J PELUSO
• 金额: $ 16.42万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-15 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7867760
• 关键词: Agonist; Apoptosis; Binding; Binding Proteins; Cell Differentiation process; Cell Line; Cell Survival; Cells; Cholesterol; Data; Decidual Cell Reactions; Development; Endometrial; Endometrium; Epithelial Cell Proliferation; Estrogens; Estrous Cycle; Female; Fertility; Gene Expression; Goals; Gonadotropins; Grant; Granulosa-Lutein Cells; Growth; Histocompatibility; Hormones; Human; Immune system; Immunoprecipitation; In Vitro; Infertility; Knockout Mice; Lead; Ligands; Longevity; Luteal Cells; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Mediator of activation protein; Membrane; Mesenchymal; Mitosis; Monitor; Mouse Strains; Mus; Mutation; Nuclear; Ovarian; Ovary; Ovulation; Physiological; Play; Pregnancy; Premature Ovarian Failure; Progesterone; Progesterone Receptors; Protein Binding; Proteomics; Reproduction; Reproductive Physiology; Reproductive Process; Research Personnel; Role; Site; Staging; Steroids; Stromal Cells; Technology; Testing; Tissues; Two-Dimensional Gel Electrophoresis; Uterine Cancer; Uterus; base; embryo/fetus; granulosa cell; implantation; in vivo; natural Blastocyst Implantation; public health relevance; receptor; reproductive; reproductive axis; reproductive function; transcription factor

DESCRIPTION (provided by applicant): Progesterone (P4) is an essential hormone that regulates the entire reproductive process. P4 has many target sites throughout the reproductive axis but two major sites of action include the ovary and the uterus. The ovary synthesizes and secretes P4 throughout the menstrual/estrous cycle with its levels increasing during pregnancy. P4 acts directly of the granulosa cells of the ovary to inhibit mitosis and apoptosis. In addition, the P4 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 classical nuclear P4 receptors (PGRs). However, not all of the actions of P4 can be explained by activation of PGRs, since a number of cell lines that do not express these 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). Based on these in vitro data, we hypothesize that PGRMC1 plays essential roles in regulating ovarian and uterine function in vivo. In this grant we will directly test our hypothesis by generating a conditional knockout mouse in which PGRMC1 is specifically depleted within the granulosa/luteal cells of the ovary and the mesenchymal (stromal) tissue of the uterus (Specific Aim 1). We will then use the conditional PGRMC1 knockout mouse to monitor the effects of depleting PGRMC1 on 1) female fertility, 2) ovarian function (i.e. follicular growth, steroid synthesis and ovulation) and 3) uterine function (i.e. endometrial decidualization and embryo implantation) (Specific Aim 2). In Specific Aim 3 we will isolate PGRMC1 binding proteins by immunoprecipitation using lysates of ovarian and uterine tissues and then using two- dimensional gel electrophoresis and subsequent proteomics to identify PGRMC1 binding partners. The successful completion of the proposed studies will provide compelling evident to support PGRMC1's role in female reproductive physiology. What is important about establishing PGRMC1 as a mediator of specific ovarian and uterine functions is that this will allow for the development of a new class of P4 antagonists and agonists. These putative PGRMC1 modulators would be completely different from the present-day PGR antagonists and would only interfere with the actions of P4 that are mediated via PGRMC1 and not the PGR. These putative PGRMC1 modulators could find application in the treatment of human fertility and potentially ovarian cancers, since PGRMC1 promotes the viability of these cancers. PUBLIC HEALTH RELEVANCE: Progesterone (P4) is an essential hormone that regulates the entire female reproductive process. 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). In this grant we will conclusively determine the role of PGRMC1 by generating a conditional knockout mouse and then monitoring the effects of depleting PGRMC1 on 1) female fertility, 2) ovarian function and 3) uterine function. If successful, the proposed studies could lead to the development of a new class of P4 antagonists and agonists that could be used to treat human infertility and potentially ovarian and uterine cancers.

描述（由申请人提供）：孕酮（P4）是调节整个生殖过程的必需激素。 P4在整个生殖轴上都有许多目标部位，但两个主要作用部位包括卵巢和子宫。卵巢在整个月经/发情周期中综合并分泌P4，其在怀孕期间的水平升高。 P4直接作用于卵巢的颗粒细胞，以抑制有丝分裂和凋亡。此外，P4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合成。 P4在卵巢和子宫内的许多作用都是由经典的核P4受体（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，因为许多未表达这些受体的细胞系以及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。基于这些体外数据，我们假设PGRMC1在调节体内卵巢和子宫功能中起着重要作用。在这笔赠款中，我们将通过产生条件基因敲除小鼠直接检验我们的假设，其中PGRMC1在卵巢的颗粒/黄体细胞中特异性耗尽，并在子宫内的间充质（基质）组织（特异性AIM 1）。然后，我们将使用条件的PGRMC1基因敲除小鼠来监测耗尽PGRMC1对1）女性生育能力的影响，2）卵巢功能（即卵泡生长，类固醇合成和排卵）和3）子宫功能（即心层型脱骨和胚胎植入和胚胎植入）（特定目标）（特定目标）2）。在特定目标3中，我们将使用卵巢和子宫组织的裂解物进行免疫沉淀来分离PGRMC1结合蛋白，然后使用两维凝胶电泳和随后的蛋白质组学鉴定PGRMC1结合伴侣。拟议的研究的成功完成将为支持PGRMC1在女性生殖生理学中的作用提供明显的说服力。将PGRMC1作为特定卵巢和子宫功能的中介者建立重要的是，这将允许发展新的P4拮抗剂和激动剂。这些推定的PGRMC1调节器将与当今的PGR拮抗剂完全不同，并且只会干扰通过PGRMC1而不是PGR介导的P4的作用。这些推定的PGRMC1调节剂可以在治疗人类生育能力和潜在的卵巢癌中找到应用，因​​为PGRMC1促进了这些癌症的生存能力。 公共卫生相关性：孕酮（P4）是一种调节整个女性生殖过程的重要激素。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。在这笔赠款中，我们将通过产生有条件的基因敲除小鼠，然后监测耗尽PGRMC1对1）女性生育能力，2）卵巢功能和3）子宫功能来最终确定PGRMC1的作用。如果成功的话，拟议的研究可能会导致一类新的P4拮抗剂和激动剂的发展，这些拮抗剂和激动剂可用于治疗人类不育症以及潜在的卵巢癌和子宫癌。