Chromatin-mediated alternative splicing in reward pathophysiology

奖赏病理生理学中染色质介导的选择性剪接

基本信息

批准号：
10188481
负责人：
Elizabeth A Heller
金额：
$ 48.3万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10188481
关键词：
Address Alternative Splicing Behavior Brain Brain region Cell Culture System Chromatin Chronic DNA Data Data Set Engineering Enzymes Epigenetic Process Functional disorder Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Transcription Histone H3 Histones Learning Lysine Mediating Mental Depression Mental disorders Methylation Modification Molecular Mus Nature Neurobiology Neurons Nuclear Pathology Pharmaceutical Preparations Post-Translational Protein Processing Prevalence Protein Isoforms Proteins Publishing RNA Splicing Rewards Role Self Administration Spliced Genes Stimulus Structure Testing Volition Work addiction brain reward regions chromatin modification chromatin remodeling cocaine exposure epigenome gene function genome-wide innovation interdisciplinary approach learned behavior mRNA Precursor motivated behavior novel strategies response transcription factor transcriptome sequencing

项目摘要

PROJECT SUMMARY The epigenome refers to covalent modifications of nuclear histone proteins and their associated DNA, which function to regulate gene expression. There is a wealth of data implicating epigenetic remodeling in neurobiological function and behavior, especially in the context of reward pathologies, such as addiction and depression. However, due to the promiscuity of the factors involved, previous studies have failed to distinguish between the mere presence and the functional relevance of a given chromatin modification at a specific gene. This limits the elucidation of the precise molecular mechanisms by which neuroepigenetic remodeling regulates transcription. To address this, we utilize a multidisciplinary approach that involves (1) analysis of chromatin immunoprecipiation (ChIP)- and RNA-sequencing (seq) datasets from brain reward regions following volitional reward behavior and (2) direct manipulation of such modifications, using an innovative strategy of gene-targeted epigenetic editing using engineered transcription factors (ETFs). The current proposal tests the hypothesis that histone posttranslational modifications (HPTMs), specifically histone H3 lysine 36 methylation (H3K36me3), directly functions in reward-mediated pre-mRNA alternative splicing. The rationale for this hypothesis includes recently published data that both alternative splicing and H3K36me3 enrichment are highly regulated by cocaine exposure, and our preliminary finding that there is a significant correlation between genome-wide H3K36me3 enrichment and the splicing complexity of expressed alternative isoforms. While chromatin-mediated alternative splicing is well established in cell- culture systems, it has not yet been described as a mechanism in brain, despite the prevalence of both widespread chromatin remodeling and alternative splicing in neurons. This proposal outlines a novel strategy to demonstrate neuronal H3K36me3-mediated alternative splicing, and the functional significance of this transcriptional mechanism to motivated behavior. In particular, we will analyze neuronal changes in mouse brain reward regions following drug or natural reward self-administration. This work will establish a strategy through which we can examine additional mechanisms of chromatin-mediated alternative splicing in various brain regions, expanding beyond the initial hypotheses of the current proposal.

项目摘要表观基因组是指核组蛋白及其的共价修饰相关的DNA，该DNA功能调节基因表达。有很多数据暗示表观遗传重塑在神经生物学功能和行为中，尤其是在奖励病理的背景，例如成瘾和抑郁。但是，由于涉及因素的滥交，以前的研究未能区分给定染色质修饰在特定基因上的存在和功能相关性。这限制了神经概性的精确分子机制的阐明重塑调节转录。为了解决这个问题，我们利用一种多学科的方法涉及（1）分析染色质免疫沉淀（CHIP）和RNA-Sequist（SEQ）自愿奖励行为和（2）直接来自大脑奖励区域的数据集使用靶向基因的表观遗传学的创新策略来操纵这种修饰使用工程转录因子（ETF）进行编辑。当前的提案检验了组蛋白翻译后修饰的假设（HPTMS），特别是组蛋白H3赖氨酸36甲基化（H3K36ME3），直接在奖励介导的前MRNA替代剪接。该假设的理由包括最近发布的数据，替代剪接和H3K36me3富集都很高受可卡因暴露调节，我们的初步发现存在显着相关性全基因组H3K36ME3富集与表达的剪接复杂性之间替代同工型。虽然染色质介导的替代剪接在细胞中已经很好地确定培养系统，尽管存在普遍存在，但尚未被描述为大脑中的一种机制在神经元中的广泛染色质重塑和替代剪接的两种。这个建议概述了一种新的策略，以证明神经元H3K36ME3介导的替代剪接，并且这种转录机制对动机行为的功能意义。尤其，我们将分析药物或天然后小鼠脑奖励区域的神经元变化奖励自我管理。这项工作将建立一种我们可以检查的策略染色质介导的替代剪接的其他机制在各个大脑区域，扩展超出了当前建议的最初假设。