Vascular Estrogen Receptor Signaling Pathways

血管雌激素受体信号通路

• 批准号: 7822204
• 负责人: RICHARD H KARAS
• 金额: $ 1.59万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7822204
• 关键词: Animal Welfare; Bibliography; Binding; Biological Assay; Biology; Blood Vessels; Cardiovascular system; Cell membrane; Cells; Co-Immunoprecipitations; Complex; Consensus; Country; Data; Development; Endothelial Cells; Environment; Environmental Impact; Equipment; Estrogen Receptors; Estrogens; Event; Family member; Figs - dietary; Gene Expression; Genes; Genetic Transcription; Genomics; Grant; Hormones; Hospitals; Hypertension; IACUC; In Vitro; International; Laboratories; Left; Ligands; Literature; Manuscripts; Mediating; Methods; Mining; Molecular; Mus; Nuclear Hormone Receptors; Pathway interactions; Peptides; Phenotype; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Physiology; Principal Investigator; Proteins; Published Comment; Receptor Signaling; Regulation; Reporter; Reporting; Research; Research Ethics Committees; Resources; Response Elements; Role; Scaffolding Protein; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Suggestion; Tail; Tertiary Protein Structure; Testing; Therapeutic Agents; Time; Transcription Coactivator; Transcriptional Activation; Transcriptional Regulation; Transfection; Transgenic Mice; Vascular Endothelial Cell; Vascular System; Vertebrates; abstracting; activating transcription factor; activator protein 3; base; clinically relevant; cohort; expiration; human NOS3 protein; human RIPK1 protein; human subject; in vivo; insight; interest; knock-down; member; mutant; non-genomic; novel; novel therapeutics; prevent; programs; protein complex; receptor; response

Estrogen receptors (ERs) mediate the diverse effects of estrogen on the cardiovascular system. The two known ERs, ER1 and ER2, are ligand-activated transcription factors. The transcriptional effects of ERs depend on receptor interacting proteins, including SRC3, a member of the family of co-activator proteins. Commonly, the genomic effects just described are differentiated from more rapid effects that do not require alterations in gene expression and thus are called `non-genomic' effects. The best-studied non-genomic ER pathway in vascular endothelial cells (EC) involves rapid activation of the kinase Akt, and subsequent activation of endothelial nitric oxide synthase (eNOS). eNOS protein abundance is also transcriptionally regulated by ERs. These two signaling pathways, genomic and non-genomic, have largely been considered independent, and the physiologic relevance of the non-genomic pathway has been questioned. We recently identified the protein striatin as an ER1 interacting protein necessary for assembly of cell membrane-associated signaling complexes that mediate non-genomic estrogen-induced eNOS phosphorylation. Initially, we demonstrated that inhibition of ER1-striatin binding blocked rapid activation or Akt and eNOS, but did not inhibit transcriptional activation of an estrogen-response-element driven reporter, supporting that the striatin-ER1 interaction is not required for transcriptional activation by ER1. However, we now report that inhibition of ER1-striatin binding prevents ER1-mediated transcriptional regulation of the eNOS gene, and that the co-activator protein SRC3 in necessary for both non-genomic and genomic regulation of eNOS by ER1. Based on these findings, we propose the central hypothesis of this application, that a multi-protein complex that includes ER1, the scaffolding protein striatin, and the coactivator SRC3 regulates both non-genomic and genomic effects of estrogen in EC and vascular physiology. We propose to test this hypothesis with the following specific aims: Specific Aim 1: To investigate the mechanisms by which striatin and SRC3 regulate rapid (non-genomic) ER-dependent activation of eNOS in ECs, Specific Aim 2: To investigate the mechanisms by which striatin and SRC3 regulate ER-dependent transcriptional (genomic) activation of eNOS in ECs, and Specific Aim 3: To determine the importance of ER-striatin binding in vivo by studying a transgenic mouse that expresses a peptide that disrupts ER1-striatin binding. The results of these studies will provide insight into the importance of non-genomic ER1-mediated signaling in vascular cells and its role in regulating longer-term genomic effects of the receptor. These studies have important implications for furthering our understanding of the effects of estrogen on the vascular system and for developing novel therapeutic agents that act via these signaling pathways.

雌激素受体（ERS）介导雌激素对心血管系统的各种影响。两个已知的ER ER1和ER2是配体激活的转录因子。 ERS的转录作用取决于受体相互作用的蛋白质，包括SRC3，这是共激活蛋白家族的成员。通常，仅描述的基因组效应与不需要基因表达改变的更快作用有所不同，因此称为“非基因组”效应。血管内皮细胞（EC）中最佳研究的非基因组ER途径涉及激酶AKT的快速激活，以及随后激活内皮一氧化氮合酶（ENOS）。 ENOS蛋白丰度也受ER的转录调节。这两种信号通路（基因组和非基因组）在很大程度上被认为是独立的，并且已经质疑非基因组途径的生理相关性。我们最近将蛋白质纹状体鉴定为ER1相互作用的蛋白质，用于组装与细胞膜相关的信号传导复合物，介导非基因组雌激素诱导的eNOS磷酸化。最初，我们证明了对ER1--延纹蛋白结合的抑制阻塞了快速激活或AKT和eNOS，但没有抑制雌激素反应元素驱动的记者的转录激活，这支持了ER1转录激活的Striatin-ER1相互作用。但是，我们现在报告说，抑制ER1-间替蛋白结合可以防止ER1介导的ENOS基因的转录调控，并且共激活蛋白SRC3对于非基因组和基因组调节ER1所必需的必要条件。基于这些发现，我们提出了该应用的中心假设，即包括ER1在内的多蛋白络合物，脚手架蛋白纹状体以及共激活剂SRC3调节雌激素在EC和血管生理学中的非基因组和基因组效应。 We propose to test this hypothesis with the following specific aims: Specific Aim 1: To investigate the mechanisms by which striatin and SRC3 regulate rapid (non-genomic) ER-dependent activation of eNOS in ECs, Specific Aim 2: To investigate the mechanisms by which striatin and SRC3 regulate ER-dependent transcriptional (genomic) activation of eNOS in ECs, and Specific Aim 3: To determine the importance of ER-striatin通过研究表达破坏ER1-瑞蒂蛋白结合的肽的转基因小鼠，在体内结合。这些研究的结果将洞悉血管细胞中非基因组ER1介导的信号传导的重要性及其在调节受体长期基因组效应中的作用。这些研究对我们进一步了解雌激素对血管系统的影响以及开发通过这些信号通路起作用的新型治疗剂具有重要意义。