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Cortical organoid models to study autism-associated 16p.11.2.CNV

用于研究自闭症相关 16p.11.2.CNV 的皮质类器官模型

基本信息

批准号：
10537569
负责人：
LILIA M IAKOUCHEVA
金额：
$ 70.63万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10537569
关键词：
16p 16p11.2 ATAC-seq Address Affect Biological Bipolar Disorder Body Size Body mass index Brain Cells Cerebrum Chromatin Closure by clamp Copy Number Polymorphism Data Defect Development Diagnosis Disease Electrophysiology (science)Fibroblasts Future Gene Expression Genes Genotype Head Heterogeneity Human Immunofluorescence Immunologic Individual Intellectual functioning disability Intervention Studies Macrocephaly Measurement Mental disorders Microcephaly Modeling Molecular Morphology Mus Neurites Neurodevelopmental Disorder Neurons Obesity Organoids Patch-Clamp Techniques Pathway interactions Patients Pharmacology Phenotype Population Property Quantitative Reverse Transcriptase PCR Reporting Resolution Risk Factors Sampling Schizophrenia Signal Pathway Slice Stains Synapses Synaptic Transmission Techniques Testing Time WNT Signaling Pathway XCL1 gene autism spectrum disorder brain size brain volume cell type dosage high body mass index induced pluripotent stem cell insight migration mouse model multi-electrode arrays nerve stem cell neuronal excitability patch clamp relating to nervous system rho single-cell RNA sequencing transcriptomics

项目摘要

SUMMARY The 16p11.2 Copy Number Variant (CNV) containing 29 genes represents one of the strongest risk factors for neurodevelopmental disorders. It is the most common among all rare CNVs implicated in Autism Spectrum Disorder (ASD). Both, deletions (DEL) and duplications (DUP) of this locus are strongly associated with ASD, whereas DEL are also associated with intellectual disability, and DUP are also associated with schizophrenia. The 16p11.2 CNV dosage has “mirror” effect on the brain volume and body mass index (BMI) in human carriers diagnosed with ASD. Macrocephaly and high BMI are observed in the DEL carriers, and microcephaly and low BMI are observed in the duplication carriers. Surprisingly, 16p11.2 DEL and DUP mouse models have brain and body size phenotypes opposite to human carriers. Despite the importance of this CNV, the detailed molecular mechanisms that are disrupted during early brain development remain unknown. We recently generated induced pluripotent stem cells (iPSCs) and cerebral organoids from fibroblasts of 16p11.2 DEL and DUP patients with extreme head size phenotypes. Cerebral organoids recapitulated patients’ brain size phenotypes. Excess neuron number and depletion of neuroprogenitors was observed in DEL, with a “mirror” phenotypes in DUP. In addition, neuron migration and synaptic defects were identified, implicating Rho/Wnt signaling. The central hypothesis of this proposal is that that 16p11.2 CNV dosage (DEL or DUP) impacts brain development by altering the ratio of different neuronal (sub)types and by perturbing their electrophysiological properties. To test this hypothesis, we will: (1) Investigate how 16p11.2 CNV impacts cell type populations and chromatin accessibility at the single cell resolution; (2) Investigate how 16p11.2 CNV affects synaptic transmission and oscillatory network activity in organoids using whole-cell patch-clamp and multielectrode array (MEA) recordings; Investigate involvement of Rho/Wnt and other signaling pathways by rescuing molecular and cellular phenotypes of DEL and DUP organoids. Our study has a potential to uncover new mechanisms behind 16p11.2 CNV in autism.

总结包含29个基因的16p11.2拷贝数变异体（CNV）是最强的风险因素之一，神经发育障碍它是自闭症谱系中所有罕见的CNV中最常见的自闭症（ASD）。该基因座的缺失（DEL）和重复（DUP）均与ASD密切相关，而DEL也与智力残疾有关，DUP也与精神分裂症有关。 16p11.2 CNV剂量对人类携带者的脑体积和体重指数（BMI）具有“镜像”效应被诊断为ASD。在DEL携带者中观察到大头畸形和高BMI，在DEL携带者中观察到小头畸形和低BMI。 BMI在重复基因携带者中存在。令人惊讶的是，16p11.2 DEL和DUP小鼠模型具有脑和体型表型与人类携带者相反。尽管这种CNV的重要性，在早期大脑发育过程中被破坏的机制仍然未知。我们最近产生了诱导多能干细胞（iPSC）和来自16p11.2 DEL和DUP患者的成纤维细胞的脑类器官，极端头部大小表型。脑类器官概括了患者的脑大小表型。过剩神经元在DEL中观察到神经祖细胞的数量和消耗，在DUP中观察到“镜像”表型。此外，本发明还提供了一种方法，鉴定了神经元迁移和突触缺陷，涉及Rho/Wnt信号传导。核心假设 16p11.2 CNV剂量（DEL或DUP）通过改变大脑发育的方式影响大脑发育。不同神经元（子）类型的比例，并通过干扰它们的电生理特性。测试基于这一假设，我们将：（1）研究16p11.2 CNV如何影响细胞类型群体和染色质（2）研究16p11.2 CNV如何影响突触传递，使用全细胞膜片钳和多电极阵列（MEA）记录的类器官中的振荡网络活动; 通过拯救分子和细胞表型研究Rho/Wnt和其他信号通路的参与 DEL和DUP类器官。我们的研究有可能揭示16p11.2 CNV背后的新机制。自闭症