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：
10383778
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/10383778
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

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

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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.

项目摘要/摘要孕酮受体膜成分（PGRMC）1和PGRMC2被认为介导孕酮的作用。我们的实验室最近对鼠PGRMC1和PGRMC2基因进行了努力，以评估这些基因在女性生育的背景下的功能。使用PGR-CRE小鼠的诱变研究表明在每个基因的条件消融中，PGRMC1和PGRMC2对于女性生育至关重要这已经发展为过早的生殖衰老。尽管是孕酮受体，但子宫内膜PGRMC1表达实际上是在增殖性雌二醇（E2）二世相中最高的人类和灵长类动物的月经周期。 PGRMC家族的进化起源早于将固醇作为信号分子的出现至少3亿年。因此，现在变得清晰 PGRMC具有孕激素依赖性和孕激素独立的功能。评估 PGRMC1D/D，PGRMC2D/D和PGRMC1/2D/D小鼠中的雌激素反应显示PGRMC蛋白是 E2诱导的子宫内膜上皮细胞增殖根本上需要。此外，我们确定在人子宫内膜癌中，PGRMC1表达升高。与这些发现一致源自PGRMC1过表达细胞的子宫内膜异种移植肿瘤生长速度更快，并且更具耐药性进行化学疗法治疗。我们实验室中最近的蛋白质组学努力已经确定PGRMC1与三个主要蛋白质组，其中包括糖酵解酶，RNA结合蛋白和蛋白质参与翻译的启动。众所周知，雌激素会诱导子宫内膜WARBURG样糖酵解作用。我们的中心假设是PGRMC家庭成员有助于协调E2诱导的子宫内膜细胞增殖通过与RNA结合蛋白的相互作用，并通过建立类似Warburg的有氧糖酵解状态确保有足够的合成碳基建筑材料可用于增殖和扩展在月经/发情循环的增殖阶段的组织。该假设的链接组成部分 PGRMC家庭成员调节有利于Warburg样糖酵解的mRNA加工。通过使用蛋白质组学，原代细胞培养物，旨在评估子宫内膜上皮细胞的小鼠模型体内增殖，新型转基因小鼠的发展以及RNA-Seq/clip-seq分析，该假设将在以下特定目的中进行测试：1）评估PGRMC1过表达的后果女性生育能力以及子宫内膜增生和癌症的发展； 2）证明PGRMC1 通过在子宫内膜上皮中建立类似Warburg的糖酵解作用来促进E2诱导的增殖细胞； 3）证明PGRMC1至少介导了子宫内膜中E2的一些增殖作用上皮细胞通过其与RNA结合蛋白的相互作用处理和易位到多核糖体。这些机械研究的结果将提供新颖有关PGRMC1功能的分子细节的信息，这些函数可能有用可以减少E2驱动子宫内膜疾病的患病率和/或严重程度的对策。