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、内部和外部风险因素都会影响未成熟的大脑，导致认知和认知能力下降。行为缺陷发生在很晚的时期，症状通常不会在成年之前延迟出现。因此，纵向建模在神经精神疾病的背景下至关重要。然而，即使之后迄今为止已发表数百项研究，神经系统的功能基因组学仍然面临着巨大的挑战障碍：实际上目前所有可用的转录组和表观基因组方法都是横截面的，仅提供组织收获时间点的基因组功能快照。该领域急需使追求“回顾性功能基因组学”成为可能的工具箱。这将允许，对于例如，动物成年后的行为与其神经元（或神经胶质）的状态直接相关生命早期的基因组。该探索性提案将开发绘制神经元基因组图谱的技术纵向组织，从幼年和年轻成年小鼠暴露于风险开始与认知功能长期损害相关的因素，包括亚慢性治疗 NMDA 受体拮抗剂药物与社会隔离压力。我们将绘制 3 维基因组图谱皮质神经元特定亚型的组织和功能，包括兴奋性投射神经元和抑制性快速尖峰中间神经元，并开发绘制神经元基因组组织图的技术纵向背景。我们的回顾性 3D 基因组作图方法基于短暂的、基于疱疹的在前额皮质中表达细菌 Dam 甲基腺嘌呤嵌合结构的载体甲基转移酶，与锚定于顺式调控序列的合成 DNA 结合蛋白融合神经元基因。其中包括编码 GABA 合成酶的 Gad1、编码活性调节酶的 Arc 细胞骨架蛋白和编码脑源性神经营养因子的 Bdnf。我们的方法是回顾性的 3D 基因组图谱将使我们能够直接将动物的行为和认知与空间联系起来三个月前风险暴露期间神经元基因组的结构。这这里提出的方法如果成功的话，可以为探索分子和与长期个体间变异相关的机制的神经表观遗传基础对早期生活经历和童年逆境的适应能力。