Adolescent Binge Ethanol Dysregulation of H3K36me3 Leads to Cryptic Transcription and Lasting Memory Deficits

青少年暴饮乙醇 H3K36me3 失调导致隐性转录和持久记忆缺陷

• 批准号: 10569496
• 负责人: Emily Rose Brocato
• 金额: $ 3.24万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-25 至 2023-05-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10569496
• 关键词: Adolescence; Adolescent; Adult; Affect; Alcohol consumption; Alcohols; Animal Behavior; Animals; Attention; Automobile Driving; Behavior; Behavioral; Binding; Bioinformatics; Biological Assay; Brain; Candidate Disease Gene; ChIP-seq; Chromatin; Cognitive; Cognitive deficits; Critical Thinking; DNA-Directed RNA Polymerase; Data; Deposition; Development; Dose; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Ethanol; Exposure to; Future; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Targeting; Genes; Genetic Transcription; Genomic Segment; Genomics; Goals; Histones; Investigation; Knowledge; Lead; Length; Lysine; Male Adolescents; Malignant Neoplasms; Measures; Mediating; Memory; Memory impairment; Methylation; Modeling; Molecular; Mus; Nucleosomes; Pathology; Pharmacologic Substance; Play; Prefrontal Cortex; Quantitative Reverse Transcriptase PCR; Reporting; Research; Research Personnel; Role; Stimulus; Techniques; Time; Tissue-Specific Gene Expression; Tissues; Training; Transcript; Transcriptional Regulation; Untranslated RNA; Viral Vector; Visuospatial; Water; Writing; adolescent alcohol exposure; adolescent binge drinking; alcohol exposure; behavioral outcome; chromatin immunoprecipitation; chromatin remodeling; cognitive change; cohort; differential expression; epigenetic regulation; epigenome; gene product; histone methylation; histone methyltransferase; indexing; long term memory; memory consolidation; novel; object recognition; overexpression; prevent; promoter; skill acquisition; skills; success; transcriptome sequencing; underage drinking

PROJECT SUMMARY The prefrontal cortex (PFC) undergoes significant changes during adolescence, and alcohol exposure during this time is particularly damaging. Consuming alcohol, especially in binges, has been shown to negatively impact the adolescent brain, resulting in brain structural changes and lasting deficits in attention, visuospatial skills, and memory. The underlying mechanisms of these deficits are largely unknown; however, histone epigenetic regulation of gene expression provides an attractive mechanism for driving long-term behavioral and cognitive changes following binge ethanol. Through interaction with environmental stimuli, such as alcohol, epigenetic alterations can determine downstream gene expression, which in turn affects behavioral outcomes. The proposed study will investigate whether histone epigenetic alterations are the mechanism behind the persistent memory deficits of adolescents exposed to binge levels of ethanol. Specifically, the study will focus on tri-methylated histone 3, lysine 36 (H3K36me3). H3K36me3 has been shown to be affected by adolescent binge ethanol, and also plays significant roles in transcription regulation and memory function. These functional roles may provide a mechanism for how dysregulation of H3K36me3 by adolescent binge ethanol could underlie long-term memory deficits. Additionally, H3K36me3 is an under-studied mark, and further investigation of its role in memory function and cryptic transcription will provide valuable data for others involved in behavioral epigenetic research. Through this proposal, we intend to determine how H3K36me3-regulated loci and gene expression patterns within the PFC are altered after adolescent binge ethanol. By overlaying chromatin immunoprecipitation (ChIP) sequencing with RNA sequencing data, we will determine how ethanol- induced dysregulation of H3K36me3 affects gene expression, and at which loci these changes are occurring. We also plan to manipulate H3K36me3 levels within the PFC to determine if altering this epigenetic mark can change the behavioral outcomes of adolescent binge ethanol. These studies therefore have the potential to expand the knowledge of how epigenetic alterations can induce lasting behavioral changes. The success of these epigenetic and genomic studies will identify novel genes that might contribute to the persistent memory deficits associated with adolescent binge ethanol, which could ultimately lead to pharmaceutical targets to reverse cognitive deficits associated with adolescent binge drinking. Additionally, this study will facilitate my training in molecular techniques, bioinformatics analyses, and animal behavior models. In addition to the critical thinking, scientific writing, and professional development skills that I will be trained in, this research will prepare me to become a successful independent researcher studying how environmental influences, such as alcohol consumption, is encoded into the epigenome and how this can cause lasting impacts to behavior.

项目摘要 前额叶皮层（PFC）在青春期经历了显着的变化， 这一次特别具有破坏性。饮酒，特别是在狂欢中，已被证明是负面的， 影响青少年的大脑，导致大脑结构变化和注意力、视觉空间 技能和记忆这些缺陷的潜在机制在很大程度上是未知的;然而，组蛋白 基因表达的表观遗传调控为驱动长期行为提供了一种有吸引力的机制， 和认知能力的改变通过与环境刺激的相互作用，如酒精， 表观遗传改变可以决定下游基因表达，这反过来又影响行为结果。 这项拟议的研究将调查组蛋白表观遗传改变是否是 青少年暴露于过量酒精的持续记忆缺陷。具体而言，研究将侧重于 三甲基化组蛋白3，赖氨酸36（H3 K36 me 3）。H3 K36 me 3已被证明是受青少年 酗酒，并在转录调控和记忆功能中发挥重要作用。这些功能 作用可能提供了一种机制，即青少年酗酒如何导致H3 K36 me 3失调， 造成长期记忆缺失此外，H3 K36 me 3是一个研究不足的标记， 它在记忆功能和神秘转录中的作用将为其他参与研究的人提供有价值的数据。 行为表观遗传学研究通过这项提议，我们打算确定H3 K36 me 3调控的基因座 青少年酗酒后PFC内的基因表达模式发生改变。通过叠加 染色质免疫沉淀（ChIP）测序与RNA测序数据，我们将确定如何乙醇- H3 K36 me 3的诱导失调影响基因表达，以及这些变化发生在哪些位点。 我们还计划操纵PFC中的H3 K36 me 3水平，以确定改变这种表观遗传标记是否可以 改变青少年酗酒的行为结果。因此，这些研究有可能 扩展表观遗传改变如何引起持久的行为变化的知识。的成功 这些表观遗传学和基因组学的研究将确定新的基因，可能有助于持久的记忆。 与青少年酗酒有关的缺陷，这可能最终导致药物目标， 逆转与青少年酗酒相关的认知缺陷。此外，这项研究将有助于我 分子技术、生物信息学分析和动物行为模型方面的培训。除了有 批判性思维，科学写作和专业发展技能，我将接受培训，这项研究将 准备我成为一个成功的独立研究人员研究如何环境的影响，如 酒精消费，被编码到表观基因组中，以及这如何对行为造成持久的影响。