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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.

项目总结/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，大约1%的美国儿童。ASD的特征是言语交流的缺陷，社交互动和有限的重复性兴趣ASD患者的一个子集显示早期大脑过度生长。我们已经建立了两个相关的模型，模拟ASD早期大脑过度生长的重要方面： Dvl 1和Dvl 3缺陷模型（Dvl 1-/-3+/-突变体），显示成人社会行为异常与深层皮质形成期间短暂的胚胎脑扩大有关; 由源自以下的诱导多能干细胞（iPSC）产生的神经元祖细胞（NPC）模型：与非ASD相比，早期脑过度生长的ASD个体显示出增强的增殖对照值得注意的是，在这两种模型中，这些异常的ASD表型是由β- 连环蛋白活性及其直接靶点BRN 2。我们假设β-catenin/BRN 2转录级联反应是一种一个在脑发育过程中精细调节NPC增殖和分化的关键途径，小鼠和人类，导致正常的社会行为，而失调导致异常的社会行为以及ASD的至少某些方面。我们建议解决这个假设所提出的关键问题。目的1：鉴定来自来源于人的iPSC的NPC中β-连环蛋白和BRN 2的转录靶点。具有和不具有早期脑过度生长的人类患者以及来自Dvl 1-/-3+/-突变小鼠。以确定 β-catenin/BRN 2转录级联反应在伴有早期脑过度生长的ASD中的特异性，我们将产生来自iPSC的NPC：对照组和有和无早期脑过度生长的ASD患者; PTEN患者显示有或无ASD的大头畸形的突变;以及两个具有校正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）分析来鉴定在来自野生型和非野生型小鼠NPC中由β-catenin和BRN 2直接调节的下游途径， Dvl 1-/-3+/-胚胎和来源于对照和ASD患者的iPSC的人NPC。目的2：表征人类ASD中错误调节的常见下游途径，凌乱的小鼠模型。我们将交叉下游途径，介导β- 连环蛋白/BRN 2转录级联，并选择通常使用新的生物信息学工具和方法。目的3：在由以下物质制成的NPC和类器官中，证实目的2中鉴定的途径的破坏来自对照和ASD患者以及Dvl 1-/-3+/-小鼠的胚胎脑的iPSC。我们会确认，验证目标2中确定的通路的中断，并评估其在ASD病理学中的重要性，通过基因组编辑在我们的小鼠和人类来源的NPC和脑类器官模型中干扰它们。将通过基因组编辑产生具有强效应的基因/途径的小鼠突变体，以验证其有效性。在脑过度生长和体内社会行为中的意义。