UNDERSTANDING THE CONSEQUENCES OF SLEEP LOSS IN AN AUTISM MOUSE MODEL

项目摘要 睡眠不足在神经发育障碍中很常见，如自闭症谱系障碍（ASD）， 93%的ASD患者报告有睡眠问题。这些问题恶化了生活质量和核心症状， ASD可能先于ASD诊断，这表明它们在生命早期就开始了。的潜在不利影响 早期生活睡眠中断得到动物模型研究的支持，这些研究显示了对 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 在出生后第24天（P24）更加突出，Wnt单一途径优先下调 与成年相比。然而，转录反应和分子机制的基础上， 在ASD的背景下，对SD在生命早期的不利影响了解甚少。我的总体目标是了解 睡眠不足对神经发育障碍的有害影响。实验室先前的工作表明， 携带ASD基因Shank 3突变的小鼠（Shank 3突变C小鼠）是唯一的ASD小鼠模型， 复制了ASD患者失眠的临床特征，并显示出对SD的异常转录反应， 成年人了在我的研究生工作中，我用多导睡眠描记器记录年轻的Shank 3小鼠， 在P24存在并可检测到。在F99阶段，我将研究这种ASD突变如何改变 SD在P24的转录上使用批量和单核RNA测序。此外，我将使用 针对SHANK 3和β-连环蛋白的靶向定量蛋白质组学研究Shank 3 β-连环蛋白C和SD对 典型的Wnt信号传导。核β-连环蛋白水平是经典Wnt信号传导的公认标志物。 激活，并且SHANK 3已显示结合细胞核中的β-连环蛋白并抑制其转录活性。 我的赞助商，Peixoto博士有一个既定的跟踪记录分析转录组反应SD小鼠 模型，并建立了ASD睡眠研究的Shank 3模型。我的共同赞助人弗兰克医生 在发育神经科学方面拥有丰富的经验，并开创了许多研究， 睡眠在发育性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 和普拉萨德博士是单细胞转录组学和蛋白质组学的世界级专家。在博士预科F99阶段， 我将学习使用蛋白质组学和转录组学方法和先进的转录组学数据分析。在 博士后K 00阶段，我将建立在这些技能和使用遗传小鼠模型，以集成电路的水平 操纵和多组学，以更好地了解睡眠不足对行为的后果， 神经发育障碍总的来说，拟议的培训将使我能够开始一个独立的 在领先的神经科学研究机构的研究生涯，并推进我们对分子的理解， 睡眠不足的后果在整个发展。

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.