权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Functional characterization of an enhancer RNA in cerebral ischemia

增强子 RNA 在脑缺血中的功能表征

基本信息

批准号：
10118570
负责人：
Ashutosh Dharap
金额：
$ 36.03万
依托单位：
JFK MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-30 至 2021-09-24
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10118570
关键词：
Acetylation Acute Adult Affect Binding Binding Sites Biological Biological Assay Brain Brain Injuries Budgets Cell Adhesion Cell Cycle Cell Death Cerebral Ischemia Cerebral cortex ChIP-seq Chromatin Cyclic AMP DNA DNA Polymerase II Data Development Disease Down-Regulation Enhancers Estrogens Extracellular Matrix Female Fluorescent in Situ Hybridization Functional disorder Future Gene Expression Gene Expression Regulation Gene Proteins Genes Genetic Enhancer Element Genetic Transcription Genomics Histones Immunoprecipitation Infarction Investigation Ischemia Ischemic Stroke Lysine Maps Mediating Methods Middle Cerebral Artery Occlusion Molecular Molecular Target Mus Neurologic Neurons Outcome Pathologic Pathway interactions Phase Phenotype Physiological Play Proteins RNA Regulator Genes Reperfusion Therapy Reporter Reporting Role Running Sensorimotor functions Signal Transduction Stimulus Stroke Structure TP53 gene Testing Therapeutic Time Transcript Translations Ubiquitination Untranslated RNA Western Blotting Work base blood-brain barrier disruption cdc Genes cell type crosslink endoplasmic reticulum stress experimental study genome-wide genomic locus improved in vivo knock-down loss of function macrophage male new therapeutic target novel post stroke programs response sex stroke outcome transcription factor transcriptome sequencing virtual

项目摘要

Recent studies have reported widespread transcription of mammalian enhancers into noncoding RNA transcripts in a stimulus-dependent manner. Growing evidence shows that these enhancer RNAs (eRNAs) have essential roles in orchestrating higher-order chromatin interactions to facilitate gene expression and phenotypic outcomes during development and disease. As a result, eRNAs are emerging as an important component of the gene regulatory machinery. Due to their very recent discovery, the expression and roles of eRNAs in stroke is virtually unknown. Recently, we applied a combination of genome-wide RNA-seq and genome-wide enhancer mapping using ChIP-seq to identify a number of stroke-induced eRNAs at multiple time-points of reperfusion in the mouse cerebral cortex. Our preliminary data confirmed enhancer activity of the genomic loci encoding the eRNAs, showed that the eRNAs are localized to the chromatin, and revealed an important role for one such eRNA in modulating post-stroke brain damage and gene expression. The molecular interactions, functional mechanisms and sex-dependent effects of eRNAs on the post-stroke pathophysiology are unexplored. In the current project, we build upon our preliminary data to evaluate eRNA functionality in-depth during stroke in the adult mouse cortex. Specific Aim 1 will use a combination of crosslinking-immunoprecipitation, fluorescence in situ hybridization and transcriptional-state analysis to determine the molecular targets of the eRNAs as a function of post-stroke reperfusion time in the mouse cortex. Specific Aim 2 will employ eRNA loss-of-function in vivo followed by cellular, physiological, pathological and neurological analyses to evaluate the role of the eRNA in propagating the post-stroke pathophysiology. Specific Aim 3 will evaluate the molecular targets and pathophysiological effects of the eRNA in the female mouse cortex. Together, this work will illuminate the significance of eRNAs in the cerebral cortex and may reveal novel gene regulatory relationships in stroke. This work will pave the way for future studies exploring the therapeutic manipulation of eRNAs to improve post-stroke outcomes.

最近的研究报道了哺乳动物增强子广泛转录成非编码的rna。以依赖刺激的方式进行转录。越来越多的证据表明，这些增强子RNA(ERNA) 在协调高阶染色质相互作用中发挥重要作用，以促进基因表达和发育和疾病期间的表型结果。因此，eRNAs正在成为一种重要的基因调控机制的组成部分。由于他们最近的发现，它的表达和作用中风中的eRNAs几乎是未知的。最近，我们应用了全基因组RNA-seq和使用CHIP-SEQ进行全基因组增强子作图以识别多个卒中诱导的eRNA 小鼠大脑皮层再灌流的时间点。我们的初步数据证实了该蛋白的增强活性编码eRNAs的基因组位点，表明eRNAs定位于染色质，并揭示了一个这样的Erna在调节中风后脑损伤和基因表达方面发挥了重要作用。这个 ERNAs在卒中后的分子相互作用、作用机制及性别依赖效应病理生理学尚未被研究。在当前的项目中，我们基于我们的初步数据来评估Erna 中风期间成年小鼠大脑皮质功能的深入研究。《特定目标1》将结合使用交联免疫沉淀、荧光原位杂交和转录状态分析确定eRNAs的分子靶点作为小鼠卒中后再灌注时间的函数大脑皮层。具体目标2将在体内使用Erna功能丧失，随后是细胞、生理、病理以及神经学分析，以评估ERNA在传播卒中后病理生理方面的作用。特异性目标3将评估ERNA在女性中的分子靶点和病理生理效应小鼠皮质。总之，这项工作将阐明eRNAs在大脑皮层中的意义，并可能揭示中风中新的基因调控关系。这项工作将为未来探索对eRNAs的治疗操作以改善中风后的预后。