PGRMC1 function in female reproductive physiology

PGRMC1 在女性生殖生理学中的功能

基本信息

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
项目类别：
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

项目摘要

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）：黄体酮（P4）是调节整个生殖过程的必需激素。 P4 在整个生殖轴上有许多靶位点，但两个主要作用位点包括卵巢和子宫。卵巢在整个月经/动情周期合成和分泌 P4，其水平在怀孕期间增加。 P4 直接作用于卵巢的颗粒细胞，抑制有丝分裂和细胞凋亡。此外，P4 还能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经典核 P4 受体 (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，因为许多不表达这些受体的细胞系以及 Pgr 缺失小鼠能够对 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）。基于这些体外数据，我们假设 PGRMC1 在体内调节卵巢和子宫功能中发挥重要作用。在这笔资助中，我们将通过生成条件性基因敲除小鼠来直接检验我们的假设，其中 PGRMC1 在卵巢的颗粒/黄体细胞和子宫的间充质（基质）组织中被特异性地耗尽（具体目标 1）。然后，我们将使用条件性 PGRMC1 敲除小鼠来监测 PGRMC1 耗尽对 1) 女性生育力、2) 卵巢功能（即卵泡生长、类固醇合成和排卵）和 3) 子宫功能（即子宫内膜蜕膜化和胚胎着床）的影响（具体目标2)。在具体目标 3 中，我们将使用卵巢和子宫组织裂解物通过免疫沉淀分离 PGRMC1 结合蛋白，然后使用二维凝胶电泳和随后的蛋白质组学来鉴定 PGRMC1 结合伴侣。拟议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令人信服的证据。将 PGRMC1 确立为特定卵巢和子宫功能调节剂的重要之处在于，这将允许开发一类新的 P4 拮抗剂和激动剂。这些假定的 PGRMC1 调节剂与目前的 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。这些假定的 PGRMC1 调节剂可用于治疗人类生育能力和潜在的卵巢癌，因为 PGRMC1 可以促进这些癌症的生存能力。公众健康相关性：黄体酮 (P4) 是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）。在这笔资助中，我们将通过生成条件性基因敲除小鼠，然后监测耗尽 PGRMC1 对 1) 女性生育能力、2) 卵巢功能和 3) 子宫功能的影响，最终确定 PGRMC1 的作用。如果成功，拟议的研究可能会导致开发一类新的 P4 拮抗剂和激动剂，可用于治疗人类不孕症以及潜在的卵巢癌和子宫癌。