Enhancer RNA Regulation of Experience-dependent Neuroepigenetic Processes

经验依赖性神经表观遗传过程的增强子 RNA 调节

• 批准号: 9893018
• 负责人: JEREMY J DAY
• 金额: $ 43.72万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893018
• 关键词: Adaptive Behaviors; Adult; Behavior; Behavioral; Binding; Biological Assay; Biological Models; Biology; Brain; Brain Diseases; CRISPR/Cas technology; Cells; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Cognition Disorders; DNA; DNA Methylation; DNA Sequence; Development; Disease; Distal Enhancer Elements; Electrophoretic Mobility Shift Assay; Enhancers; Ensure; Epigenetic Process; Expression Profiling; Future; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Generations; Genes; Genetic Enhancer Element; Genetic Transcription; Genomics; High-Throughput Nucleotide Sequencing; Hippocampus (Brain); Histone Acetylation; Histones; Immediate-Early Genes; Immunoprecipitation; Individual; Investigation; Learning; Link; Lysine; Maps; Memory; Mental Health; Mental disorders; Mitotic; Modeling; Modification; Molecular; Nature; Neuronal Plasticity; Neurons; Parents; Pattern; Phenotype; Physiology; Play; Process; RNA; RNA delivery; RNA-Protein Interaction; Regulation; Regulator Genes; Regulatory Element; Rodent; Role; Signal Transduction; Specificity; Synaptic plasticity; System; Testing; Therapeutic; Tissue-Specific Gene Expression; Transcript; Treatment Efficacy; Untranslated RNA; Variant; base; behavioral plasticity; brain health; cognitive process; complex biological systems; conditioned fear; crosslink; experience; experimental study; falls; gene induction; genome sequencing; genome-wide; insight; knock-down; mammalian genome; neuron development; neuropsychiatric disorder; novel; novel strategies; novel therapeutics; programs; promoter; response; screening; single molecule; therapeutic candidate; tool; transcription factor; whole genome

Distal enhancer elements in DNA enable higher-order chromatin interactions that facilitate gene expression programs and thus contribute to cellular phenotype and function. Enhancers regulate numerous aspects of cell and tissue-specific gene expression patterns in the developing and adult brain, and have been highly implicated in mental health and disease. In neuronal systems, enhancer elements are subject to widespread, bidirectional transcription that yields non-coding enhancer RNA (eRNA). However, the precise function of eRNAs is still unclear, with different models proposing unique regulatory functions. Emerging evidence suggests that eRNAs function to modify epigenetic states at gene regulatory elements, which are critical for long-term behavioral and neuronal plasticity. Specific Aim 1 of this proposal seeks to utilize whole-genome sequencing approaches to define transcriptional and epigenetic signatures of neuronal activity at enhancers. In addition to revealing new therapeutic candidates, this aim will employ novel targeted eRNA delivery strategies based on CRISPR technology to allow manipulation of eRNAs at specific enhancers to determine their function. Specific Aim 2 will examine interactions between eRNAs and epigenetic profiles at enhancers and promoters of linked genes, which will establish the hierarchical relationships between molecular interactions at enhancers. Specific Aim 3 will examine the contributions of eRNAs to neuronal function and memory formation in the adult brain using hippocampus-dependent contextual fear conditioning, a robust assay of learning and memory that requires activity-dependent gene transcription. This novel approach will demonstrate the necessity of unique eRNAs at specific genes for neuronal activity and behavior, and also enable the first investigation of whether modulation of eRNAs is sufficient to alter memory function. In addition to revealing the exact nature and scope of eRNAs in neuronal gene regulation, this proposal will pave the way for future experiments that will explore how manipulation of enhancers could be used to reprogram circuits that have become maladaptive in mental illness and cognitive disease states.

DNA中的远端增强子元件使更高级别的染色质相互作用促进基因表达 程序，从而对细胞表型和功能做出贡献。增强剂调节细胞的多个方面 和组织特异的基因表达模式，在发育中的和成年的大脑中，并且已经高度 与心理健康和疾病有牵连。在神经系统中，增强子元件受到广泛的影响， 产生非编码增强子RNA(ERNA)的双向转录。然而，它的精确功能 ERNAs仍然不清楚，不同的模型提出了独特的调控功能。新出现的证据 提示eRNAs的功能是改变基因调控元件的表观遗传状态，这对 长期的行为和神经元可塑性。这项提案的具体目标1寻求利用全基因组 确定增强子上神经元活动的转录和表观遗传特征的测序方法。在……里面 除了揭示新的治疗候选药物外，该目标还将采用新的靶向Erna给药策略 基于CRISPR技术，允许在特定增强子上操纵eRNAs以确定其 功能。具体目标2将检查eRNA和表观遗传学特征之间的相互作用在增强子和 连锁基因的启动子，这将建立分子相互作用之间的等级关系 增强剂。具体目标3将研究eRNAs对神经元功能和记忆形成的贡献 在使用海马体依赖的情景恐惧条件反射的成人大脑中，对学习和 需要依赖活动的基因转录的记忆。这一新颖的方法将展示 神经元活动和行为的特定基因上独特的eRNAs的必要性，也使第一 研究eRNA的调制是否足以改变记忆功能。除了揭示 ERNAs在神经元基因调控中的确切性质和范围，这一建议将为未来铺平道路 将探索如何使用增强剂的操作来重新编程具有 在精神疾病和认知疾病状态下变得不适应。