Regulation of endometrial proliferation by the PGRMC family

PGRMC 家族对子宫内膜增殖的调节

基本信息

批准号：
10211171
负责人：
James K Pru
金额：
$ 32.51万
依托单位：
UNIVERSITY OF WYOMING
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10211171
关键词：
Ablation Adenomyosis Aerobic Anabolism Appearance Binding Proteins Carbon Cell Proliferation Cells Deltastab Development Disease Endometrial Endometrial Carcinoma Endometrial Hyperplasia Endometrium Ensure Enzymes Epithelial Epithelial Cell Proliferation Epithelial Cells Estradiol Estrogens Estrous Cycle Evaluation Event Failure Family Family member Female Gene Expression Genes Genetic Transcription Glucose Glycolysis Gonadal Steroid Hormones Human In Vitro Infertility Leiomyoma Link LoxP-flanked allele Malignant neoplasm of cervix uteri Malignant neoplasm of ovary Measures Mediating Membrane Menorrhagia Menstrual cycle Messenger RNA Metabolic Metabolism Molecular Mus Mutagenesis Outcome Output Oxidative Stress Oxygen Phase Physiological Polyribosomes Prevalence Primary Cell Cultures Primates Progesterone Progesterone Receptors Proteins Proteomics RNA Splicing RNA-Binding Proteins Regulation Reproductive Process Resistance Severities Signal Transduction Signaling Molecule Sterols Testing Tissue Expansion Tissues Transcription Process Transgenic Mice Translation Initiation Tumor-Derived Uterus aerobic glycolysis base building materials chemotherapy crosslinking and immunoprecipitation sequencing early pregnancy endometriosis estrogenic experimental study female fertility female sex hormone gene function implantation in vivo mRNA Expression member model design mouse model novel overexpression premature proliferative phase Menstrual cycle protein function receptor reproductive senescence reproductive tract response stem subfertility transcriptome sequencing tumor xenograft uterine receptivity

项目摘要

Project Summary/Abstract Progesterone receptor membrane component (PGRMC) 1 and PGRMC2 are thought to mediate progesterone actions. Our lab recently floxed the murine Pgrmc1 and Pgrmc2 genes in an effort to evaluate the function of these genes in the context of female fertility. Mutagenesis studies using Pgr-Cre mice revealed that Pgrmc1 and Pgrmc2 are essential for female fertility in that conditional ablation of each gene results in subfertility that progresses to premature reproductive senescence. Despite being a purported progesterone receptor, endometrial PGRMC1 expression is actually highest during the proliferative, estradiol (E2)-dominated phase of the menstrual cycle in humans and primates. The evolutionary origin of the PGRMC family predates the appearance of sterols as signaling molecules by at least 300 million years. As such, it is now becoming clear that PGRMCs have both progesterone-dependent and progesterone-independent functions. An evaluation of estrogenic responses in Pgrmc1d/d, Pgrmc2d/d, and Pgrmc1/2d/d mice revealed that PGRMC proteins are fundamentally required for E2-induced endometrial epithelial cell proliferation. Furthermore, we determined that PGRMC1 expression is elevated in human endometrial cancer. Consistent with these findings, human endometrial xenograft tumors derived from PGRMC1 over-expressing cells grow faster and are more resistant to chemotherapy treatment. Recent proteomic efforts in our lab have established that PGRMC1 interacts with three principal groups of proteins, and these include glycolytic enzymes, RNA-binding proteins and proteins involved in the initiation of translation. Estrogen is known to induce an endometrial Warburg-like glycolytic effect. Our central hypothesis is that PGRMC family members help coordinate E2-induced endometrial cell proliferation through their interactions with RNA-binding proteins and by establishing a Warburg-like aerobic glycolytic state to ensure that sufficient anabolic carbon-based building materials are available for proliferation and expansion of the tissue during the proliferative phase of the menstrual/estrous cycle. A linked component of this hypothesis is that PGRMC family members regulate mRNA processing that favors Warburg-like glycolysis. Through the use of proteomics, primary cell cultures, mouse models designed for evaluating endometrial epithelial cell proliferation in vivo, development of a novel transgenic mouse, and RNA-seq/CLIP-seq analysis, this hypothesis will be tested in the following Specific Aims: 1) evaluate the consequences of PGRMC1 over-expression on female fertility and development of endometrial hyperplasia and cancer; 2) demonstrate that PGRMC1 contributes to E2-induced proliferation by establishing a Warburg-like glycolytic effect in endometrial epithelial cells; and 3) demonstrate that PGRMC1 mediates at least some of the proliferative actions of E2 in endometrial epithelial cells through its interactions with RNA-binding proteins that coordinate post-transcriptional mRNA processing and translocation to the polyribosome. The outcome of these mechanistic studies will provide novel information about the molecular details of PGRMC1 functions that may be useful in the development of countermeasures that could reduce the prevalence and/or severity of E2-driven endometrial diseases.

项目概要/摘要黄体酮受体膜成分 (PGRMC) 1 和 PGRMC2 被认为可介导黄体酮的作用。我们的实验室最近对小鼠 Pgrmc1 和 Pgrmc2 基因进行了固定，以评估这些基因在女性生育能力中的功能。使用 Pgr-Cre 小鼠进行的诱变研究表明 Pgrmc1 和 Pgrmc2 对于女性生育能力至关重要，因为每个基因的条件性消除都会导致生育能力低下进而发展为生殖早衰。尽管是一种所谓的黄体酮受体，子宫内膜 PGRMC1 表达实际上在雌二醇 (E2) 主导的增殖期最高。人类和灵长类动物的月经周期。 PGRMC 家族的进化起源早于甾醇作为信号分子的出现至少有 3 亿年。因此，现在已经很清楚了 PGRMCs 具有黄体酮依赖性和黄体酮非依赖性功能。的评价 Pgrmc1d/d、Pgrmc2d/d 和 Pgrmc1/2d/d 小鼠的雌激素反应表明 PGRMC 蛋白 E2 诱导的子宫内膜上皮细胞增殖所必需的。此外，我们确定 PGRMC1 表达在人类子宫内膜癌中升高。与这些发现一致的是，人类来自 PGRMC1 过表达细胞的子宫内膜异种移植肿瘤生长更快且更具抵抗力进行化疗治疗。我们实验室最近的蛋白质组研究已经证实 PGRMC1 与三类主要蛋白质，包括糖酵解酶、RNA 结合蛋白和蛋白质参与翻译的启动。已知雌激素可诱导子宫内膜 Warburg 样糖酵解作用。我们的中心假设是 PGRMC 家族成员有助于协调 E2 诱导的子宫内膜细胞增殖通过它们与 RNA 结合蛋白的相互作用并建立类似 Warburg 的有氧糖酵解状态确保有足够的合成代谢碳基建筑材料可用于扩散和扩张月经/动情周期的增殖期的组织。该假设的一个相关组成部分 PGRMC 家族成员调节有利于 Warburg 样糖酵解的 mRNA 加工。通过使用蛋白质组学、原代细胞培养、用于评估子宫内膜上皮细胞的小鼠模型体内增殖、新型转基因小鼠的开发以及 RNA-seq/CLIP-seq 分析，该假设将在以下具体目标中进行测试：1）评估 PGRMC1 过度表达对女性生育能力以及子宫内膜增生和癌症的发育； 2) 证明 PGRMC1 通过在子宫内膜上皮中建立类似 Warburg 的糖酵解作用，促进 E2 诱导的增殖细胞； 3)证明PGRMC1至少介导E2在子宫内膜中的一些增殖作用上皮细胞通过与协调转录后 mRNA 的 RNA 结合蛋白相互作用加工并易位至多核糖体。这些机制研究的结果将提供新颖的有关 PGRMC1 功能的分子细节的信息可能有助于开发可以降低 E2 驱动的子宫内膜疾病的患病率和/或严重程度的对策。