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.

项目摘要新生儿啮齿动物暴露于高水平的外源雌激素导致子宫异常和肿瘤以及成年后的其他病理。暴露于合成雌激素的女性中发生了类似的影响子宫中的二乙基苯甲醇（DES），因此确定早期雌激素暴露的作用具有临床意义。雌激素作用主要由雌激素受体1（ESR1）介导。大多数ESR1是核的，但5-10％是位于细胞膜中。早期雌激素暴露引起的表观遗传变化可能是随之而来的病理。早期雌激素暴露产生表观遗传异常的机制是不清楚，但可能涉及通过膜ESR1介导的信号通路（MESR1）。总体该提议的目的是建立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。为此，我们将使用两只转基因小鼠：仅核雌激素缺乏MESR1和仅膜雌激素受体（MOER）小鼠缺乏NESR1的受体（NOER）小鼠以及野生型（WT）和ESR1敲除（ESR1KO）对照。在AIM 1，WT，Noer，Moer和Esr1ko中女性将注射DES [1 mg/kg；产后天（PND）1-5]或车辆。 PND的子宫5幼崽将检查了组蛋白甲基化和乙酰化标记，这些标记也与表观遗传基因调节有关作为激酶途径和甲基转移酶复合酶催化亚基的表达一个组蛋白甲基化标记。 AIM 1中的其他实验将确定DES的表观遗传效应是否完全是通过ESR1介导的，或可能涉及其他雌激素受体，例如ESR2或G蛋白偶联的雌激素受体（GPER）。表观遗传效应是可逆的，因此在AIM 2中，我们将使用染色质免疫沉淀（芯片）和定量PCR（QPCR），以确定个体的组蛋白修饰的永久变化如AIM 1中的PND 1-5治疗的小鼠成年子宫中的雌激素反应基因。我们将检查键三个雌激素反应性的调节和非调节区域的甲基化和乙酰化位点已经报道了新生儿雌激素治疗后的组蛋白修饰的基因，并且看起来也由于新生儿治疗的结果，这些基因的反应性在成年人中具有过度反应性。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作用 WT和NOER小鼠中靶基因的位点以及反应性的反应性，将需要Mesr1，并且是BE 在表达NESR1但缺乏MESR1（不）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，这些作用也需要NESR1。这些实验的结果将增加我们对MESR1角色的理解，以及该受体如何与NESR1相互作用以调节表观遗传变化。这些结果还将增加对早期雌激素暴露会改变靶基因中组蛋白甲基化和乙酰化的机理基础，以及调节组蛋白甲基化的信号事件的级联反应；这些结果将具有临床标志性。

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.