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.

项目概要/摘要自闭症谱系障碍（ASD）是一种复杂且异质的神经发育疾病，影响在美国，大约 1% 的自闭症谱系障碍儿童的特点是言语交流障碍。一部分自闭症谱系障碍患者表现出早期大脑过度生长。我们制作了两个相关模型来模拟自闭症患者早期大脑过度生长的重要方面：小鼠缺乏 Dvl1 和 Dvl3（Dvl1-/-3+/- 突变体）的模型，表现出成人社会行为异常与深层皮质形成过程中短暂的胚胎脑增大和人类有关；由诱导多能干细胞 (iPSC) 产生的神经元祖细胞 (NPC) 模型与非自闭症谱系障碍患者相比，早期大脑过度生长的自闭症谱系障碍个体的增殖能力增强值得注意的是，在这两种模型中，这些异常的 ASD 表型都是由 β- 的下调引起的。我们发现 β-catenin/BRN2 转录级联是大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，而失调则导致异常的社会行为我们建议至少解决 ASD 的某些方面。目标 1：鉴定源自 iPSC 的 NPC 中 β-catenin 和 BRN2 的转录靶标患有和不患有早期大脑过度生长的人类患者以及 Dvl1-/-3+/- 突变小鼠进行测定。 β-连环蛋白/BRN2转录级联在患有早期大脑过度生长的自闭症谱系障碍中的特异性，我们将产生来自以下 iPSC 的 NPC：对照组和患有或不患有 PTEN 早期脑过度生长的 ASD 患者；显示患有或不患有 ASD 的大头畸形的突变；以及具有正确 Wnt 通路的两个 ASD 系我们将利用RNA-seq和ChIP-seq结合基因本体（GO）分析来识别突变。野生型和小鼠 NPC 中直接受 β-catenin 和 BRN2 调节的下游通路 Dvl1-/-3+/- 胚胎和源自对照和 ASD 患者 iPSC 的人类 NPC。目标 2：描述人类自闭症谱系障碍 (ASD) 和我们将交叉介导 β- 效应的下游途径。小鼠和人类 NPC 中的连环蛋白/BRN2 转录级联，并选择常见的途径使用新颖的生物信息学工具和方法失调。目标 3：验证目标 2 中确定的 NPC 和类器官中途径的破坏我们将确认来自对照和 ASD 患者的 iPSC 以及 Dvl1-/-3+/- 小鼠的胚胎大脑。验证目标 2 中确定的途径的破坏并评估其在 ASD 病理学中的重要性通过基因组编辑在小鼠和人源性 NPC 以及脑类器官模型中扰乱它们。通过基因组编辑产生具有强烈影响的基因/通路的小鼠突变体，以验证其在大脑过度生长和体内社会行为中具有重要意义。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Autism-specific PTEN p.Ile135Leu variant and an autism genetic background combine to dysregulate cortical neurogenesis.

自闭症特异性 PTEN p.Ile135Leu 变异和自闭症遗传背景相结合，导致皮质神经发生失调。

DOI：
发表时间：
2023-05-04
期刊：
影响因子：
9.8
作者：
Fu, Shuai;Bury, Luke A D;Eum, Jaejin;Wynshaw
通讯作者：
Wynshaw

Disruption of CTNND2, encoding delta-catenin, causes a penetrant attention deficit disorder and myopia.

编码 δ-连环蛋白的 CTNND2 的破坏会导致渗透性注意力缺陷障碍和近视。

DOI：
发表时间：
2020-10-22
期刊：
影响因子：
0
作者：
Adegbola, Abidemi;Lutz, Richard;Nikkola, Elina;Strom, Samuel P;Picker, Jonathan;Wynshaw
通讯作者：
Wynshaw

PTPN4 germline variants result in aberrant neurodevelopment and growth.

PTPN4 种系变异导致神经发育和生长异常。

DOI：
发表时间：
2021-07-08
期刊：
影响因子：
0
作者：
Chmielewska, Joanna J;Burkardt, Deepika;Granadillo, Jorge Luis;Slaugh, Rachel;Morgan, Shamile;Rotenberg, Joshua;Keren, Boris;Mignot, Cyril;Escobar, Luis;Turnpenny, Peter;Zuteck, Melissa;Seaver, Laurie H;Ploski, Rafal;Dziembowska, Magdalena;W
通讯作者：
W

Neuronal lineage tracing from progenitors in human cortical organoids reveals novel mechanisms of human neuronal production, diversity, and disease.

人类皮质类器官祖细胞的神经元谱系追踪揭示了人类神经元产生、多样性和疾病的新机制。