Membrane estrogen receptor 1 mediation of epigenetic effects of estrogen

膜雌激素受体1介导雌激素的表观遗传效应

基本信息

批准号：
9401825
负责人：
Paul S. Cooke
金额：
$ 1.62万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9401825
关键词：
Acetylation Adult Affect Amino Acids Animal Model Catalytic Domain Cell Nucleus Cell membrane Cells Clinical Complex DNA DNA Methylation DNA Packaging DNA Sequence DNA-Directed RNA Polymerase Development Diethylstilbestrol Disease susceptibility ESR1 gene ESR2 gene Endocrine Endocrinology Epigenetic Process Estrogen Nuclear Receptor Estrogen Receptor alpha Estrogen Receptors Estrogens Event Exposure to Female GPER gene Gene Expression Regulation Gene Targeting Genes Genetic Transcription Goals Heritability Histone Acetylation Histone H3 Histones Human Individual Infertility Injectable Knock-out Lactoferrin Ligands Longevity Lysine Mediating Mediation Membrane Methylation Methyltransferase Modification Molecular Mus Neonatal Neoplasms Nuclear Nucleosomes Organ Pathologic Pathologic Processes Pathology Pathway interactions Phosphotransferases Physiological Physiology Predisposition Process Proteins Reporting Reproductive Health Research Rodent Role Signal Pathway Signal Transduction Site Testing Tissues Transcriptional Activation Transgenic Mice Transgenic Model Translating Uterus Woman Work calbindin chromatin immunoprecipitation chromatin modification clinically significant estrogenic experimental study female reproductive system gene function gene repression health data histone methylation histone modification imprint in utero insight mouse model neonatal exposure neoplastic novel postnatal pup receptor reproductive reproductive development reproductive function reproductive organ response tool transcription factor xenoestrogen

项目摘要

Project Summary Exposure of neonatal rodents to high levels of exogenous estrogen leads to uterine abnormalities and neoplastic and other pathologies in adulthood. Similar effects occur in women exposed to the synthetic estrogen diethylstilbestrol (DES) in utero, so determining effects of early estrogen exposure has clinical significance. Estrogen effects are mediated primarily by estrogen receptor 1 (ESR1). Most ESR1 is nuclear, but 5-10% is located in cell membranes. Epigenetic changes induced by early estrogen exposure may be major factors in ensuing pathologies. Mechanisms by which early estrogen exposure produces epigenetic abnormalities are unclear, but may involve signaling pathways mediated through membrane ESR1 (mESR1). The overall objective of this proposal is to establish roles of mESR1 and nuclear ESR1 (nESR1) in uterine epigenetic effects of early estrogen treatment. To accomplish this, we will use two transgenic mice: nuclear-only estrogen receptor (NOER) mice lacking mESR1 and membrane-only estrogen receptor (MOER) mice lacking nESR1, along with wild-type (WT) and Esr1 knockout (Esr1KO) controls. In Aim 1, WT, NOER, MOER and Esr1KO females will be injected with DES [1 mg/kg; postnatal days (PND) 1-5] or vehicle. Uteri of PND 5 pups will examined for histone methylation and acetylation marks critically involved in epigenetic gene regulation, as well as the kinase pathway and expression of the catalytic subunit of the methyltransferase complex responsible for one histone methylation mark. Other experiments in Aim 1 will determine if epigenetic effects of DES are entirely mediated through ESR1, or could involve other estrogen receptors such as ESR2 or G protein-coupled estrogen receptor (GPER). Epigenetic effects are reversible, so in Aim 2 we will use chromatin immunoprecipitation (ChIP) and quantitative PCR (qPCR) to determine permanent changes in histone modifications of individual estrogen-responsive genes in adult uteri of mice treated on PND 1-5, as in Aim 1. We will examine key methylation and acetylation sites across regulatory and non-regulatory regions of three estrogen-responsive genes for which histone modifications following neonatal estrogen treatment have been reported, and also look for hyperresponsiveness of these genes to estrogen in the adult as a consequence of neonatal DES treatment. Our hypothesis is that epigenetic effects of early estrogen exposure on histone methylation and acetylation sites, and hyperresponsiveness of target genes in WT and NOER mice, will obligatorily require mESR1, and be absent in mice expressing nESR1 but lacking mESR1 (NOER). In addition, we postulate that mESR1 by itself will be insufficient to mediate estrogen-induced epigenetic effects and that these effects will also require nESR1. Results of these experiments will increase our understanding of the roles of mESR1, and how this receptor interacts with nESR1 to regulate epigenetic changes. These results will also increase understanding of the mechanistic basis by which early estrogen exposure alters histone methylation and acetylation in target genes, and the cascade of signaling events regulating histone methylation; these results will have clinical signficance.

项目概要新生啮齿动物暴露于高水平的外源雌激素会导致子宫异常和肿瘤以及成年期的其他病症。接触合成雌激素的女性也会出现类似的效果己烯雌酚（DES）在子宫内产生，因此确定早期雌激素暴露的影响具有临床意义。雌激素作用主要由雌激素受体 1 (ESR1) 介导。大多数 ESR1 是核电，但 5-10% 是位于细胞膜上。早期雌激素暴露引起的表观遗传变化可能是导致随之而来的病理。早期雌激素暴露产生表观遗传异常的机制是尚不清楚，但可能涉及通过膜 ESR1 (mESR1) 介导的信号通路。整体该提案的目的是确定 mESR1 和核 ESR1 (nESR1) 在子宫表观遗传学中的作用早期雌激素治疗的影响。为了实现这一目标，我们将使用两只转基因小鼠：仅核雌激素缺乏 mESR1 受体（NOER）小鼠和缺乏 nESR1 的仅膜雌激素受体（MOER）小鼠，以及野生型 (WT) 和 Esr1 敲除 (Esr1KO) 对照。在目标 1、WT、NOER、MOER 和 Esr1KO 中雌性将注射 DES [1 mg/kg；产后天数（PND）1-5]或车辆。 PND 5 幼崽的子宫将检查与表观遗传基因调控密切相关的组蛋白甲基化和乙酰化标记，以及作为激酶途径和甲基转移酶复合物催化亚基的表达，负责 1 个组蛋白甲基化标记。目标 1 中的其他实验将确定 DES 的表观遗传效应是否完全有效。通过 ESR1 介导，或可能涉及其他雌激素受体，例如 ESR2 或 G 蛋白偶联雌激素受体（GPER）。表观遗传效应是可逆的，因此在目标 2 中我们将使用染色质免疫沉淀 (ChIP) 和定量 PCR (qPCR) 来确定个体组蛋白修饰的永久性变化如目标 1 所示，接受 PND 1-5 治疗的小鼠成年子宫中的雌激素反应基因。我们将检查关键跨三个雌激素反应性调节区和非调节区的甲基化和乙酰化位点已报道新生儿雌激素治疗后组蛋白修饰的基因，并且还查找新生儿 DES 治疗导致这些基因对成人雌激素过度反应。我们的假设是早期雌激素暴露对组蛋白甲基化和乙酰化的表观遗传影响 WT 和 NOER 小鼠中靶基因的位点和高反应性将强制需要 mESR1，并且在表达 nESR1 但缺乏 mESR1 (NOER) 的小鼠中不存在。此外，我们假设 mESR1 本身不足以介导雌激素诱导的表观遗传效应，并且这些效应也需要 nESR1。这些实验的结果将加深我们对 mESR1 作用的理解，以及该受体如何与 nESR1 相互作用来调节表观遗传变化。这些结果也将加深对早期雌激素暴露改变靶基因组蛋白甲基化和乙酰化的机制基础，以及调节组蛋白甲基化的信号事件级联；这些结果将具有临床意义。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Membrane estrogen receptor 1 is required for normal reproduction in male and female mice.

膜雌激素受体 1 是雄性和雌性小鼠正常繁殖所必需的。

DOI：
发表时间：
2017
期刊：
Journal of endocrinology and reproduction : JER
影响因子：
0
作者：
Nanjappa,ManjunathaK;Mesa,AnaM;Tevosian,SergeiG;deArmas,Laura;Hess,RexA;Bagchi,IndraniC;Cooke,PaulS
通讯作者：
Cooke,PaulS

Estrogen in the male: a historical perspective.