A novel intracellular 7TM estrogen receptor in breast

雌激素是人体内的一种重要荷尔蒙，调节许多不同的生理作用，包括 多种组织的生长、发育和动态平衡。雌激素在体内起着至关重要的作用 正常乳腺组织的发育，而对雌激素的反应性是最大的 乳腺癌患者的重要预后和治疗因素。这些属性中的许多都具有 被归因于一种可溶的雌激素受体(ER)，其主要功能是作为配体激活的 转录因子。我们已经鉴定了一种新的细胞内，7-跨膜，G蛋白- 偶联雌激素受体(GPR30)，我们提出了与ER一起调节细胞和组织的功能 对雌激素的反应性。我们发现GPR30主要存在于内质中 网状结构。目标1的目标是确定GPR30内调节其细胞位置的序列。 我们还开发了一种新的荧光雌激素衍生物家族，它既能与雌激素受体结合，又能与雌激素受体结合 GPR30。在目标2中，我们将使用这些荧光雌激素来开发基于细胞的雌激素结合分析 GPR30，并确定一系列雌激素相关化合物的结合特异性和亲和力。 最近，我们描述了乳腺癌中GPRSO介导的新的信号转导途径。 细胞。雌激素介导的磷脂酰肌醇3，4，5-三磷酸的核积聚 GPR30反式激活表皮生长因子受体(EGFR)目标3的目标是调查 与EGFR磷酸化和内化有关的EGFR反式激活的机制 导致转录激活和细胞生长控制。最后，我们建立了GPR30的殖民地 空白小鼠解剖GPR30在乳房生长发育中的体内功能。在目标4中，我们将 鉴定GPR30在野生型小鼠中的表达，并分析GPR30缺失小鼠的表型。我们会 还描述了GPR30和经典ER都缺失的小鼠的表型。 使用这些工具，我们可以解决GPR30的雌激素反应信号在基因定义的 背景资料。建议的实验对于确定这种新型雌激素的作用将是非常宝贵的- 雌激素介导的正常发育中的生长、增殖和分化中的反应性受体 在肿瘤进展和乳腺癌的治疗方面也是如此。

Estrogen is a crucial hormone in the human body, regulating many diverse physiological effects, including the growth, development and homeostasis of numerous tissues. Estrogen plays a critical role in the development of normal breast tissue, whereas responsiveness to estrogen represents one of the most important prognostic and therapeutic factors for patients with breast cancer. Many of these properties have been attributed to a soluble receptor for estrogen (ER) that functions in large part as a ligand-activated transcription factor. We have characterized a novel intracellular, 7-transmembrane spanning, G protein- coupled estrogen receptor (GPR30) that we propose functions alongside ER to regulate cell and tissue responsiveness to estrogen. We have discovered that GPR30 resides predominantly in the endoplasmic reticulum. The goal of Aim 1 is to determine the sequences within GPR30 that regulate its cellular location. We have also developed a novel family of fluorescent estrogen derivatives that binds to both the ER and GPR30. In Aim 2 we will use these fluorescent estrogens to develop cell-based estrogen binding assays for GPR30 and determine the binding specificities and affinities of a collection of estrogen-related compounds. Recently, we have characterized novel GPRSO-mediated signal transduction pathways in breast cancer cells. The nuclear accumulation of phosphatidylinositol 3,4,5-trisphosphate is initiated by estrogen-mediated transactivation of epidermal growth factor receptor (EGFR) by GPR30. The goal of Aim 3 is to investigate the mechanisms involved in EGFR transactivation with respect to EGFR phosphorylation and internalization, leading to transcriptional activation and cell growth control. Finally, we have established a colony of GPR30 null mice to dissect the in vivo function of GPR30 in breast growth and development. In Aim 4, we will characterize GPR30 expression in wild type mice and analyze the phenotypes of GPR30 null mice. We will also characterize the phenotypes of mice in which both GPR30 and the classical ERs have been deleted. With these tools, we can address estrogen-responsive signaling of GPR30 in a genetically defined background. The experiments proposed will be invaluable in defining the roles of this novel estrogen- responsive receptor in estrogen-mediated growth, proliferation, and differentiation in normal development as well as in neoplastic progression and the treatment of breast cancer.