UNDERSTANDING THE CONSEQUENCES OF SLEEP LOSS IN AN AUTISM MOUSE MODEL

了解自闭症小鼠模型中睡眠不足的后果

• 批准号: 10829011
• 负责人: Elizabeth Medina
• 金额: $ 4.77万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-18 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10829011
• 关键词: Adult; Adverse effects; Advisory Committees; Affect; Age; Animal Model; Applied Skills; Behavior; Behavioral; Binding; Brain; C-terminal; Cell Nucleus; Cells; Clinical; Cognition; Cognitive deficits; Data Analyses; Data Set; Development; Diagnosis; Disease; Down-Regulation; Early Intervention; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Models; Genetic Transcription; Genomics; Glutamates; Goals; Human; Individual; Infant; Intervention; Knowledge; Learning; Life; Long-Term Effects; Molecular; Molecular Abnormality; Mus; Mutation; Neurodevelopmental Disorder; Neurosciences; Neurosciences Research; Nuclear; Nuclear RNA; Nuclear Translocation; Pathway interactions; Phase; Phenotype; Polysomnography; Positioning Attribute; Postdoctoral Fellow; Proteomics; Publishing; Quality of life; Reporting; Repression; Research; Research Institute; Research Project Grants; Role; Scaffolding Protein; Siblings; Sleep; Sleep Deprivation; Sleep Disorders; Sleep disturbances; Sleeplessness; Small Nuclear RNA; Study models; Symptoms; Synapses; Synaptic plasticity; Syndrome; Technical Expertise; Testing; Training; WNT Signaling Pathway; Wild Type Mouse; Work; autism spectrum disorder; beta catenin; career; cell type; cognitive ability; cognitive function; excitatory neuron; experience; falls; gene repression; improved; individuals with autism spectrum disorder; insight; member; mouse model; multiple omics; mutant; neurodevelopment; poor sleep; postnatal; pre-doctoral; programs; response; single nucleus RNA-sequencing; skills; sleep quality; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Poor sleep is common in neurodevelopment disorders such as autism spectrum disorder (ASD), with up to 93% of ASD individuals reporting sleep problems. These problems worsen quality of life and core symptoms of ASD and likely precede an ASD diagnosis, suggesting they start early in life. The potential adverse impact of early life sleep disruption is supported by animal model studies showing long term functional consequences on behavior. However, little is known about the underlying molecular consequences of sleep deprivation (SD) early in life. Our recent work in wildtype mice demonstrates that the cortical transcriptional response to SD is more prominent at postnatal day 24 (P24) with a preferential downregulation on the Wnt-singling pathway compared to adulthood. Nevertheless, the transcriptional response and molecular mechanisms underlying the adverse effects of SD early in life are poorly understood in the context of ASD. My overall goal is to understand the detrimental consequences of sleep loss in neurodevelopmental disorders. Previous work in the lab showed that mice carrying a mutation in the ASD gene Shank3 (Shank3∆C mice) are the only ASD mouse model that replicates the clinical features of insomnia in ASD and show an abnormal transcriptional response to SD in adults. In my graduate work, I used polysomnography in young Shank3∆C mice to show that sleep problems are present and detectable at P24. In the F99 phase, I will investigate how this ASD mutation alters the impact of SD on transcription at P24 using both bulk and single nuclear RNA sequencing. In addition, I will use targeted quantitative proteomic for SHANK3 and -catenin to investigate the effects of Shank3∆C and SD on canonical Wnt-signaling. Nuclear -catenin levels are a well-established marker of canonical Wnt-signaling activation and SHANK3 has been shown to bind -catenin in the nucleus and inhibit its transcriptional activity. My sponsor, Dr. Peixoto has an established track record analyzing transcriptomic response to SD in mouse models and has establishes the Shank3∆C model for the study of sleep in ASD. My co-sponsor, Dr. Frank has extensive experience in developmental neuroscience and has pioneered many of the studies characterizing the role of sleep-in developmental synaptic plasticity. Additional members of my advisory committee, Dr Hicks and Dr. Prasad are world experts on single-cell transcriptomics and proteomics. In the pre-doctoral F99 phase, I will learn to use proteomic and transcriptomics approaches and advanced transcriptomic data analysis. In the postdoctoral K00 phase, I will build upon these skills and use genetic mouse models to integrate circuit level manipulation and multi-omics to better understand the consequences of sleep loss on behavior in neurodevelopmental disorders. Overall, the proposed training will optimally position me to start an independent research career at a leading neuroscience research institute and advance our understanding on the molecular consequences of sleep loss across development.

项目摘要 睡眠不良在神经发育障碍中很常见，例如自闭症谱系障碍（ASD），最多 93％的ASD个人报告睡眠问题。这些问题的生活质量更糟和核心症状 ASD并可能先于ASD诊断，这表明它们在生命的早期开始。潜在的不利影响 动物模型研究表明长期功能后果对早期生活的睡眠中断的影响 行为。但是，对于睡眠剥夺（SD）的潜在分子后果知之甚少 生命的早期。我们最近在Wildtype小鼠中的工作表明，对SD的皮质转录反应是 在产后第24天（P24）更突出，在Wnt-singling途径上首选下调 与成年相比。然而，转录响应和分子机制 在ASD的背景下，SD早期SD的不利影响很少。我的总体目标是了解 神经发育障碍中睡眠损失的有害后果。实验室的先前工作显示 在ASD基因shank3（shank3ΔC小鼠）中携带突变的小鼠是唯一的ASD小鼠模型 复制ASD失眠的临床特征，并显示出对SD异常的转录反应 成年人。在我的研究生工作中，我使用了年轻的shank3ΔC小鼠中的多个术语表明睡眠问题 在P24处存在并可以检测到。在F99阶段，我将研究ASD突变如何改变影响 使用批量和单个核RNA测序在P24处转录的SD。此外，我会使用 针对Shank3和-catenin的靶向定量蛋白质组学研究Shank3ΔC和SD的影响 典型的Wnt信号。核-catenin水平是典型的WNT信号的良好标记 激活和shank3已被证明可以在细胞核中结合-catenin并抑制其转录活性。 我的赞助商Peixoto博士拥有既定的记录记录，分析了对鼠标中SD的转录组响应 模型并已建立了用于研究ASD睡眠的Shank3ΔC模型。我的共同赞助商弗兰克博士 在发育神经科学方面的丰富经验，并开创了许多表征的研究 睡眠发育突触可塑性的作用。我咨询委员会的其他成员希克斯博士 Prasad博士是单细胞转录组学和保护剂的世界专家。在博士前F99阶段， 我将学习使用蛋白质组学和转录组方法以及高级转录组数据分析。在 博士后K00阶段，我将基于这些技能并使用遗传鼠标模型来整合电路水平 操纵和多词，以更好地了解睡眠损失对行为行为的后果 神经发育障碍。总体而言，拟议的培训将最佳地定位于我开始独立 领先的神经科学研究所的研究生涯，并促进我们对分子的理解 整个发育过程中睡眠损失的后果。