Understanding the interaction between sleep and SHANK3/B-catenin signaling in Autism

了解自闭症中睡眠与 SHANK3/B-catenin 信号传导之间的相互作用

• 批准号: 10627198
• 负责人: Lucia Peixoto
• 金额: $ 49.79万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10627198
• 关键词: Affect; Animals; Architecture; Attention; Binding; Brain; Brain region; C-terminal; Candidate Disease Gene; Caregivers; Cell Nucleus; Chromatin; Chronic; Circadian Rhythms; Cognition; Development; Developmental Process; Diagnosis; Disease; Disease model; Drowsiness; EP300 gene; Emotional; Equilibrium; Etiology; Family; Future; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genetic study; Genome; Glutamates; Hippocampus (Brain); Human; Impairment; In Vitro; Individual; Intervention; Lead; Length; Link; Memory; Modeling; Multiomic Data; Mus; Mutant Strains Mice; Mutation; Neurons; Nuclear; Nuclear Protein; Patients; Pharmacology; Phelan-McDermid syndrome; Phenotype; Positioning Attribute; Prefrontal Cortex; Problem behavior; Process; Protein Isoforms; Proteomics; Public Health; Quality of life; Resolution; Rodent; Severities; Signal Transduction; Sleep; Sleep Deprivation; Sleep Disorders; Sleeplessness; Small Nuclear RNA; Surveys; Symptoms; Synapses; Syndrome; Testing; Tissues; Validation; Wild Animals; autism spectrum disorder; base; beta catenin; chromatin remodeling; emotion regulation; experience; falls; improved; individuals with autism spectrum disorder; insight; mouse model; mutant; recruit; response; sleep abnormalities; sleep onset; targeted treatment; transcription factor; treatment group

PROJECT SUMMARY Sleep problems occur at a higher rate in Autism Spectrum Disorder (ASD) than in typical development. Problems falling asleep predict severity of ASD core symptoms and associated problem behaviors, and lead to chronic sleep deprivation (SD). Understanding the effects of sleep deprivation is important because it adversely affects cognition, attention and emotional regulation. Nonetheless, little is known about the mechanisms responsible for abnormal sleep or the consequences of chronic sleep loss in ASD. Identifying these mechanisms may lead to new interventions that will improve quality of life of patients and their families. The study of genetic syndromes with high rate of ASD diagnosis has provided important clues into mechanisms. One such syndrome is Phelan McDermid Syndrome (PMS), characterized by deletions in the SHANK3 gene. Mutations in SHANK3 are also often found in patients with idiopathic ASD. We previously showed that the ASD sleep phenotype is recapitulated in mice with a C-terminal deletion in Shank3 (Shank3∆C), which struggle to fall asleep despite being sleepy (i.e. after SD). We also showed that SD induces greater number of differences in gene expression in Shank3∆C animals than WT. Therefore, we hypothesize that Shank3∆C animals display an abnormal transcriptional response to SD. The objective of this proposal is to identify the mechanisms that underlie the abnormal response to SD in ASD, using Shank3 mutant mice as a model. Our preliminary studies show that SD increases the nuclear localization of full-length SHANK3a, an isoform that is absent in Shank3∆C animals and thought to be primarily synaptic. SHANK3 binds b-catenin. Translocation of β-catenin into the nucleus regulates transcription through chromatin remodeling and is key to many developmental processes. Our preliminary studies show that SD produces changes in chromatin topology in glutamatergic neurons, and that some of these changes do not take place in Shank3∆C animals. Therefore, our central hypothesis is that SD changes chromatin topology and gene expression by increasing nuclear SHANK3a and b-catenin and that the absence of SHANK3a in Shank3∆C interferes with this process. To test our hypothesis, in Aim 1, we will first determine the effects of SD in SHANK3a/b-catenin synaptic-nuclear signaling. Then, in Aim 2, we will determine the effect of SD in SHANK3-dependent chromatin topology and transcription at single-nuclear resolution. We will examine two brain regions important to ASD and impacted by sleep loss: pre-frontal cortex and hippocampus. We will subsequently test candidates prioritized through omic studies for their ability to influence sleep. Completion of these aims will allow us to determine how sleep interacts with SHANK3 and β-catenin to regulate chromatin and gene expression.

项目摘要 睡眠问题以自闭症谱系障碍（ASD）较高的速度比典型 发展。入睡的问题可以预测ASD核心符号的严重程度和相关的问题 问题行为，并导致长期睡眠剥夺（SD）。了解睡眠的影响 剥夺很重要，因为它会对认知，注意力和情绪调节产生不利影响。 尽管如此，对导致异常睡眠或造成的机制知之甚少 ASD慢性睡眠损失的后果。识别这些机制可能会导致新的 干预措施将改善患者及其家人的生活质量。通用研究 ASD诊断率高的综合征已为机制提供了重要的线索。一个这样的 综合征是Phelan McDermid综合征（PMS），其特征是Shank3基因中的缺失。 Shank3中的突变也经常在特发性ASD患者中发现。我们以前显示了 ASD睡眠表型在Shank3中具有C末端缺失的小鼠中概括 （shank3ΔC），它难以跌落入睡的目的地（即SD之后）。我们还证明了SD 与wt相比，在尚克3∆C动物中诱导基因表达的差异数量更多。所以， 我们假设Shank3ΔC动物对SD表现出异常的转录反应。这 该提议的目的是确定基于异常反应的机制 在ASD中进行SD，使用shank3突变小鼠作为模型。我们的初步研究表明SD 增加了全长shank3a的核定位，这是一种在shank3ΔC中不存在的同工型 动物，被认为是主要的突触。 Shank3结合B-catenin。 β-catenin的易位 进入细胞核可以通过染色质重塑调节转录，并且是许多人的关键 发展过程。我们的初步研究表明，SD会产生染色质的变化 谷氨酸能神经元中的拓扑结构，其中一些变化不会发生在shank3ΔC中 动物。因此，我们的中心假设是SD改变了染色质拓扑结构和基因 通过增加核shank3a和b-catenin的表达，并且在 Shank3ΔC在此过程中中断。为了检验我们的假设，在AIM 1中，我们将首先确定 SD在shank3a/b-catenin突触核信号传导中的影响。然后，在AIM 2中，我们将确定 SD在单核分辨率下依赖于Shank3依赖性染色质拓扑和转录的影响。 我们将检查对ASD重要的两个大脑区域，并受到睡眠损失的影响：前额叶皮层 和海马。随后，我们将通过OMIC研究优先测试候选人 影响睡眠的能力。这些目标的完成将使我们能够确定睡眠如何与 shank3和β-catenin调节染色质和基因表达。