Regulation of endometrial proliferation by the PGRMC family

PGRMC 家族对子宫内膜增殖的调节

• 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
• 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

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.

项目总结/摘要 孕激素受体膜成分（PGRMC）1和PGRMC 2被认为介导 孕酮作用。我们的实验室最近将鼠Pgrmc 1和Pgrmc 2基因固定在一起，以评估 这些基因在女性生育能力中的作用。使用Pgr-Cre小鼠的诱变研究显示， Pgrmc 1和Pgrmc 2对女性生育力至关重要，因为每个基因的条件性切除都会导致生育力低下 会导致过早的生殖衰老尽管它是一种传说中的孕酮受体， 子宫内膜PGRMC 1的表达实际上在子宫内膜增生期、雌二醇（E2）主导期最高， 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要早3亿年。因此，现在很明显 PGRMC具有黄体酮依赖性和黄体酮非依赖性功能。评价 Pgrmc 1d/d、Pgrmc 2d/d和Pgrmc 1/2d/d小鼠的雌激素反应显示， 这是E2诱导的子宫内膜上皮细胞增殖所必需的。此外，我们确定， PGRMC 1在人子宫内膜癌中的表达升高与这些发现一致，人类 来源于PGRMC 1过表达细胞的子宫内膜异种移植肿瘤生长更快， 到化疗我们实验室最近的蛋白质组学研究已经确定，PGRMC 1与 三大类蛋白质，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样糖酵解效应。 我们的中心假设是PGRMC家族成员帮助协调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以及通过建立Warburg样有氧糖酵解状态 以确保有足够的合成代谢碳基建筑材料可供扩散和扩张 在月经/发情周期的增殖期的组织。这一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、设计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， 将在以下具体目的中进行测试：1）评估PGRMC 1过表达对人乳腺癌细胞的影响。 女性生育力和子宫内膜增生和癌症的发展; 2）证明PGRMC 1 通过在子宫内膜上皮细胞中建立Warburg样糖酵解效应促进E2诱导的增殖 3）证明PGRMC 1介导E2在子宫内膜中的至少一些增殖作用， 上皮细胞通过其与RNA结合蛋白的相互作用，协调转录后mRNA 加工和转运到多核糖体。这些机制研究的结果将提供新的 关于PGRMC 1功能的分子细节的信息，可能有助于开发 这些措施可以降低E2驱动的子宫内膜疾病的患病率和/或严重程度。