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.

项目总结 睡眠不佳在神经发育障碍中很常见，如自闭症谱系障碍(Asd)，高达 93%的自闭症患者报告有睡眠问题。这些问题使生活质量恶化，核心症状 ASD可能在ASD诊断之前出现，这表明它们在生命早期就开始了。潜在的不利影响 早期睡眠中断得到了动物模型研究的支持，该研究显示了睡眠障碍的长期功能后果 行为。然而，人们对睡眠剥夺(SD)的潜在分子后果知之甚少 在生命的早期。我们最近在野生型小鼠中的工作表明，皮质对SD的转录反应是 在出生后24天更明显(P24)，Wnt-Singling途径优先下调 与成年相比。然而，转录反应和潜在的分子机制 在ASD的背景下，人们对生命早期SD的不良影响知之甚少。我的总体目标是理解 神经发育障碍中睡眠不足的有害后果。实验室之前的工作表明 携带ASD基因Shank3突变的小鼠(Shank3∆C小鼠)是唯一的ASD小鼠模型 复制ASD失眠的临床特征，并显示对SD的异常转录反应 成年人。在我的研究生工作中，我对年轻的Shank3∆C小鼠进行了多导睡眠监测，以显示睡眠问题 在P24出现并可检测到。在F99阶段，我将研究ASD突变是如何改变影响的 使用批量和单核RNA测序的方法对P24处的SD转录进行分析。此外，我还将使用 应用靶向定量蛋白质组学技术研究Shank3、-连环蛋白、∆C和SD对血管紧张素转换酶活性的影响 规范的WNT信号。核-连环蛋白水平是典型的WNT信号的成熟标志 激活和SHANK3已被证明结合在细胞核内的-连环蛋白并抑制其转录活性。 我的赞助人Peixoto博士在分析小鼠对SD的转录反应方面有既定的记录 建立了研究ASD患者睡眠的Shank3-∆-C模型。我的联合赞助人，弗兰克医生 在发育神经科学方面有丰富的经验，并开创了许多具有以下特点的研究 睡眠在发育突触可塑性中的作用。我的顾问委员会的其他成员，希克斯博士 和普拉萨德博士都是单细胞转录组和蛋白质组学方面的世界专家。在博士前F99阶段， 我将学习使用蛋白质组学和转录组学方法以及高级转录组数据分析。在 博士后K00阶段，我将在这些技能的基础上，使用遗传小鼠模型来集成电路级 操纵和多组学以更好地了解睡眠不足对行为的影响 神经发育障碍。总体而言，拟议的培训将使我处于最有利的地位，能够开始独立的 在一家领先的神经科学研究所的研究生涯，增进我们对分子的理解 发育过程中睡眠不足的后果。