Regulation of endometrial proliferation by the PGRMC family

项目摘要/摘要 孕激素受体膜组分(PGRMC)1和PGRMC2被认为是介导 黄体酮的作用。我们的实验室最近对小鼠的PGRMC1和Pgrmc2基因进行了筛选，以评估 这些基因在女性生育能力方面的功能。用PGR-Cre小鼠进行的诱变研究表明 PGRMC1和Pgrmc2是女性生育所必需的，因为有条件地去除每个基因都会导致亚生育 这会导致过早的生殖衰老。尽管据称是孕激素受体， 子宫内膜PGRMC1的表达实际上是在增殖期，以雌二醇(E2)为主 人类和灵长类动物的月经周期。PGRMC家族的进化起源早于 固醇作为信号分子的出现至少要追溯到3亿年前。因此，现在情况变得明朗起来 PGRMCs既有孕激素依赖功能，又有孕酮非依赖功能。对…的评价 Pgrmc1d/d、Pgrmc2d/d和PGRMC1/2d/d小鼠的雌激素反应表明，PGRMC蛋白是 是雌激素诱导子宫内膜上皮细胞增殖所必需的。此外，我们确定， PGRMC1在子宫内膜癌中的表达升高。与这些发现一致，人类 来源于PGRMC1高表达细胞的子宫内膜移植瘤生长更快，耐药性更强 接受化疗。我们实验室最近的蛋白质组学研究证实，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样的糖酵解作用。 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和建立类似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用于扩散和扩展 在月经/发情周期的增殖期。这一假说的关联部分 是PGRMC家族成员调控有利于Warburg样糖酵解的mRNA处理。通过使用 用于评估子宫内膜上皮细胞的蛋白质组学、原代细胞培养、小鼠模型 体内增殖，一种新的转基因小鼠的发展，以及RNA-seq/lip-seq分析，这一假设 将在以下具体目标中进行测试：1)评估PGRMC1过度表达在 女性生育力与子宫内膜增生症和癌的发展；2)证明PGRMC1 通过在子宫内膜上皮建立Warburg样的糖酵解效应促进E2诱导的增殖 细胞；3)证明PGRMC1至少在子宫内膜中介导了E2的部分增殖作用。 上皮细胞通过与RNA结合蛋白相互作用来协调转录后mRNA 加工和转位到多聚核糖体。这些机械论研究的结果将提供新的 有关PGRMC1功能的分子细节的信息，这些信息可能对开发 可降低雌激素性子宫内膜疾病的患病率和/或严重程度的对策。

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.