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A conserved transcriptional cascade involved in brain overgrowth, social behavior and autism

与大脑过度生长、社会行为和自闭症有关的保守转录级联

基本信息

批准号：
10199748
负责人：
ANTHONY J. WYNSHAW-BORIS
金额：
$ 45.76万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-02 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10199748
关键词：
3-Dimensional ASD patient Address Adult Affect Alleles Animal Model Animals Antibodies Behavior Biological Markers Biological Models Brain CTNNB1 gene Cell Differentiation process Cell Line Cell model ChIP-seq Child Communication impairment Complex DNA Data Data Set Databases Development Disease Down-Regulation Embryo Environmental Risk Factor FZD6 gene Fibroblasts Gene Expression Genes Genetic Transcription Genome Graph Heterogeneity Human Individual Information Systems Link Macrocephaly Mediating Mediator of activation protein Modeling Mus Mutant Strains Mice Mutation Ontology Organoids PTEN gene Pathology Pathway interactions Patients Phenotype Predisposition Proteins Public Domains RNA Regulation Signal Transduction Social Behavior Social Interaction Specificity Testing United States Variant autism spectrum disorder behavioral phenotyping beta catenin bioinformatics tool brain abnormalities brain overgrowth brain size disorder control exome genetic linkage genome editing genome-wide histone modification human model in silico in vivo individuals with autism spectrum disorder induced pluripotent stem cell insight interest loss of function mutation mouse development mouse model mutant nerve stem cell novel patient subsets stem cell model stem cell proliferation transcriptome sequencing

项目摘要

PROJECT SUMMARY/ABSTRACT Autism spectrum disorder (ASD) is a complex and heterogenous neurodevelopmental disease that affects about 1% of children in the United States. ASD is characterized by deficits in verbal communication, impaired social interaction, and limited, repetitive interests. A subset of patients with ASD display early brain overgrowth. We have produced two relevant models that model important aspects of early brain overgrowth in ASD: mouse models deficient for Dvl1 and Dvl3 (Dvl1-/-3+/- mutants) that display adult social behavior abnormalities associated with transient embryonic brain enlargement during deep layer cortical formation; and human neuronal progenitor cells (NPCs) models produced from induced pluripotent stem cells (iPSCs) derived from ASD individuals with early brain overgrowth that displayed enhanced proliferation compared to non-ASD controls. Remarkably, in both models, these aberrant ASD phenotypes were caused by down-regulation of β- catenin activity and its direct target BRN2. We hypothesize that the β-catenin/BRN2 transcriptional cascade is a key pathway that exquisitely regulates NPC proliferation and differentiation during brain development of mouse and human, resulting in normal social behavior, while dysregulation results in abnormal social behavior and at least some aspects of ASD. We propose to address key questions posed by this hypothesis. Aim 1: Identify the transcriptional targets of β-catenin and BRN2 in NPCs from iPSCs derived from human patients with and without early brain overgrowth and from Dvl1-/-3+/- mutant mice. To determine specificity of the β-catenin/BRN2 transcriptional cascade in ASD with early brain overgrowth, we will produce NPCs from iPSCs from: controls and ASD patients with and without early brain overgrowth; patients with PTEN mutations that display macrocephaly with or without ASD; and two ASD lines with corrected Wnt pathway mutations. We will utilize RNA-seq and ChIP-seq combined with gene ontology (GO) analysis to identify downstream pathways that are directly regulated by β-catenin and BRN2 in mouse NPCs from wild-type and Dvl1-/-3+/- embryos and in human NPCs derived from iPSCs of control and ASD patients. Aim 2: Characterize common downstream pathway(s) that are misregulated in human ASD and Dishevelled mouse models. We will intersect the downstream pathways that mediate the effects of the β- catenin/BRN2 transcriptional cascade in mouse and human NPCs, and select pathways that are commonly dysregulated using novel bioinformatics tools and approaches. Aim 3: Validate the disruption of pathways identified in Aim 2 in in NPCs and organoids made from iPSCs from control and ASD patients as well as embryonic brains of Dvl1-/-3+/- mice. We will confirm and validate the disruption of pathways identified in Aim 2 and assess their importance in ASD pathology by perturbing them in our mouse and human-derived NPCs and brain organoid models via genome editing. Mouse mutants for genes/pathways with strong effects will be produced by genome editing to validate their significance in brain overgrowth and social behavior in vivo.

项目摘要/摘要自闭症谱系障碍(ASD)是一种复杂的异质性神经发育疾病，影响美国约有1%的儿童。ASD的特点是言语交流障碍，受损社会互动和有限的、重复的兴趣。ASD患者中的一小部分显示出早期大脑过度生长。我们已经制作了两个相关的模型来模拟ASD早期大脑过度生长的重要方面：小鼠显示成人社交行为异常的Dvl1和Dvl3(Dvl1-/-3+/-突变)缺乏的模型与深层皮质形成过程中的暂时性胚胎脑增大有关；以及人类诱导多能干细胞(IPSCs)诱导的神经前体细胞(NPC)模型与非ASD相比，患有早期脑过度生长的ASD患者表现出更强的增殖控制。值得注意的是，在这两个模型中，这些异常的ASD表型是由β-1下调引起的。连环蛋白活性及其直接靶标Brn2。我们假设β-连环蛋白/Brn2转录级联是脑发育过程中精细调控鼻咽癌增殖分化的关键途径老鼠和人，导致正常的社会行为，而失调导致不正常的社会行为至少是自闭症的某些方面。我们建议解决这一假说提出的关键问题。目的1：鉴定β-连环蛋白和Brn2的转录靶点有和没有早期脑过度生长的人类患者和来自Dvl1-/-3+/-突变小鼠的患者。要确定 β-连环蛋白/Brn2转录级联在伴有早期脑过度生长的自闭症中的特异性，我们将产生来自IPSCs的神经干细胞来源：对照组和患有和不伴有早期脑过度生长的ASD患者；PTEN患者表现为患有或不伴有ASD的巨头症的突变；以及两个具有正确的Wnt途径的ASD系突变。我们将利用RNA-SEQ和CHIP-SEQ结合基因本体(GO)分析来鉴定野生型和非野生型小鼠神经前体细胞中受β-连环蛋白和Brn2直接调控的下游通路 Dvl1-/-3+/-胚胎和来源于对照组和ASD患者IPSCs的人NPC。目的2：研究人类自闭症和自闭症患者共同的下游通路(S)。蓬头垢面的老鼠模型。我们将与调节β影响的下游路径相交- 连环蛋白/Brn2在小鼠和人鼻咽癌中的转录级联反应，并选择通常使用新的生物信息学工具和方法进行失控。目标3：验证目标2中确定的通路在NPC和由以下物质制造的有机体中的中断来自对照组和ASD患者的IPSCs以及Dvl1-/-3+/-小鼠的胚胎脑。我们将确认并验证目标2中确定的通路的中断，并通过以下方式评估它们在ASD病理中的重要性通过基因组编辑在我们的小鼠和人类衍生的NPC和脑器官模型中干扰它们。将通过基因组编辑产生具有强大作用的基因/途径的小鼠突变体，以验证其在活体内大脑过度生长和社会行为的意义。