Retrospective Genomics in Mouse Prefrontal Cortex

小鼠前额皮质的回顾性基因组学

基本信息

批准号：
9147736
负责人：
Schahram Akbarian
金额：
$ 25.43万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9147736
关键词：
3-Dimensional Adenine Adolescent Adult Affect Age Animal Behavior Animals Anxiety Architecture Back Behavior Behavioral Bilateral Binding Boxing Brain Brain-Derived Neurotrophic Factor Bypass Candidate Disease Gene Childhood Chromatin Chromosomes Cognition Cognitive Cross-Sectional Studies Cytoskeletal Proteins DNA DNA Binding DNA-Binding Proteins Dam methyltransferase Data Defect Elements Enzymes Epigenetic Process Face Gene Expression Genes Genetic Genome Genome Mappings Genomic approach Genomics Glutamates Graph Harvest Herpesviridae Housing Impairment Injection of therapeutic agent Interneurons Left Life Light Link MK801 Maps Mediating Methods Methylation Methyltransferase Modeling Molecular Mus N-Methylaspartate NMDA receptor antagonist Nervous system structure Neurons Pharmaceutical Preparations Prefrontal Cortex Printing Proteins Publishing Radial Resolution Risk Risk Factors Short-Term Memory Simplexvirus Social isolation Stress Subgroup Symptoms Technology Testing Time Tissue Harvesting Transcription Coactivator Validation arm base behavioral study cell type cognitive function deep sequencing early life exposure epigenome epigenomics functional genomics gamma-Aminobutyric Acid inhibitory neuron inter-individual variation neuropsychiatric disorder phrases promoter research study resilience scaffold synthetic construct transcriptome transcriptomics treatment duration treatment group vector young adult

项目摘要

Many genetic, internal and external risk factors impact the immature brain, resulting in cognitive and behavioral deficits at much later periods, with the delayed onset of symptoms often not before adulthood. Therefore, longitudinal modeling is critical in the context of neuropsychiatric disease. However, even after hundreds of studies published to date, functional genomics in the nervous system still faces a formidable barrier: Virtually all transcriptomic and epigenomic approaches currently available are cross-sectional, providing snapshots of genome function only for the time point of tissue harvest. The field is in urgent need of a toolbox that makes it feasible to pursue `retrospective functional genomics'. This would allow, for example, direct correlation of an animal's behavior in adulthood with the status of its neuronal (or glial) genomes in early life. This exploratory proposal will develop technology to map neuronal genome organization in a longitudinal context, starting with the exposure of juvenile and young adult mice to risk factors associated with long-lasting impairments in cognitive function, including subchronic treatment with NMDA receptor antagonist drugs and social isolation stress. We will map 3-dimensional genome organization and function in specific subtypes of cortical neurons, including excitatory projection neurons and inhibitory fast-spiking interneurons, and develop technology to map neuronal genome organization in a longitudinal context. Our retrospective 3D genome mapping approach is based on transient, Herpes-based vectors to express, in prefrontal cortex, chimeric constructs of bacterial Dam methyl-adenine methyltransferase, fused to synthetic DNA binding proteins anchored to cis-regulatory sequences of neuronal genes. These include Gad1 encoding GABA synthesis enzyme, Arc encoding an activity-regulated cytoskeletal protein and Bdnf encoding brain-derived neurotrophic factor. Our approach, as retrospective 3D-genome mapping, will allow us to directly correlate the animal's behavior and cognition with the spatial architectures of its neuronal genomes during the period of risk exposure dating back 3 months prior. The approaches presented here, if successful, could offer critical opportunities to explore the molecular and neuro-epigenetic underpinnings of mechanisms associated with long-lasting inter-individual variations in resilience to early life exposures and childhood adversity.

许多遗传，内部和外部危险因素会影响未成熟的大脑，导致认知和在更晚期的行为缺陷，症状的延迟发作通常不会在成年前。因此，在神经精神疾病的背景下，纵向建模至关重要。但是，即使之后迄今为止发表的数百项研究，神经系统中的功能基因组学仍然面临着强大的障碍：实际上所有的转录组和表观基因组方法当前可用，都是横截面，仅在组织收获的时间点提供基因组功能的快照。该领域迫切需要使追求“回顾性功能基因组学”可行的工具箱。这将允许例如，动物在成年时的行为与其神经元（或神经胶质）的状态的直接相关早期的基因组。该探索性建议将开发技术来绘制神经元基因组在纵向背景下的组织，从少年和年轻小鼠的风险开始与认知功能的长期损害相关的因素，包括 NMDA受体拮抗剂和社会隔离压力。我们将绘制3维基因组皮质神经元的特定亚型的组织和功能，包括兴奋性投影神经元和抑制性快速刺激性中间神经元，并开发技术来绘制神经元基因组组织纵向背景。我们的回顾性3D基因组映射方法基于瞬态，基于疱疹在前额叶皮层中表达的载体，细菌大坝甲基 - 腺嘌呤的嵌合构建体甲基转移酶，与合成DNA结合蛋白融合，锚定在顺式调节序列神经元基因。其中包括编码GABA合成酶的GAD1，编码活动调节的电弧细胞骨架蛋白和BDNF编码脑衍生的神经营养因子。我们的方法，作为回顾 3D基因组映射将使我们能够将动物的行为和认知与空间直接相关联其神经元基因组的体系结构可追溯到三个月前。这如果成功的话，这里提出的方法可以提供探索分子和的关键机会与长期持久的个体间变化相关的机制的神经斑观基础对早期生活暴露和童年逆境的韧性。