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

• 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
• 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

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.

项目概要 表观基因组是指核组蛋白的共价修饰及其 相关DNA，其功能是调节基因表达。有丰富的数据 表明表观遗传重塑与神经生物学功能和行为有关，特别是在神经生物学功能和行为中 奖励病理学的背景，例如成瘾和抑郁。然而，由于 由于所涉及的因素杂乱，以前的研究未能区分单纯的因素 特定基因上给定染色质修饰的存在及其功能相关性。 这限制了神经表观遗传的精确分子机制的阐明 重塑调节转录。为了解决这个问题，我们采用多学科方法 涉及 (1) 染色质免疫沉淀 (ChIP) 和 RNA 测序 (seq) 分析 来自遵循意志奖励行为的大脑奖励区域的数据集和（2）直接 使用基因靶向表观遗传的创新策略来操纵此类修饰 使用工程转录因子 (ETF) 进行编辑。 当前的提案测试了组蛋白翻译后修饰的假设 (HPTM)，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 选择性剪接。该假设的基本原理包括 最近发表的数据表明，选择性剪接和 H3K36me3 富集都高度 受可卡因暴露的调节，我们的初步发现存在显着的相关性 全基因组 H3K36me3 富集和表达的剪接复杂性之间的关系 替代异构体。虽然染色质介导的选择性剪接在细胞中已得到很好的证实 尽管在文化系统中普遍存在，但它尚未被描述为大脑中的一种机制 神经元中广泛的染色质重塑和选择性剪接。这个提议 概述了一种新的策略来证明神经元 H3K36me3 介导的选择性剪接，以及 这种转录机制对动机行为的功能意义。尤其， 我们将分析药物或自然疗法后小鼠大脑奖励区域的神经元变化 奖励自我管理。这项工作将建立一个策略，通过它我们可以检查 不同大脑区域染色质介导的选择性剪接的其他机制， 超出了当前提案的最初假设。

项目成果

Recent advances in neuroepigenetic editing.

神经表观遗传编辑的最新进展。

• 发表时间: 2019
• 影响因子: 5.7
• 作者: Xu, Song;Heller, Elizabeth A
• 通讯作者: Heller, Elizabeth A

Single sample sequencing (S3EQ) of epigenome and transcriptome in nucleus accumbens.

伏隔核表观基因组和转录组的单样本测序 (S3EQ)。

• 发表时间: 2018-10-01
• 影响因子: 3
• 作者: Xu, S J;Heller, E A
• 通讯作者: Heller, E A