Gene expression profiling of IPSC derived neurons in Autism Spectrum Disorder

自闭症谱系障碍中 IPSC 衍生神经元的基因表达谱

• 批准号: 10320346
• 负责人: JOACHIM F HALLMAYER
• 金额: $ 73.73万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-06 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10320346
• 关键词: ASD patient; Accounting; Affect; Age; Architecture; Autism Diagnosis; Automobile Driving; Autopsy; Biological; Biological Assay; Biology; Brain; Calcium; California; Cell Line; Cell Survival; Cellular Assay; Collection; Complex; Copy Number Polymorphism; Data; Derivation procedure; Development; Dimensions; Disease; Exons; Frequencies; Funding; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genomics; Genotype; Glutamates; Heterogeneity; Human; Image; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Industrialization; Institutes; International; Investigation; Length; Molecular; Morphology; Neurites; Neurodevelopmental Disorder; Neuronal Differentiation; Neurons; Outcome; Pathology; Pathway Analysis; Pathway interactions; Patients; Pattern; Peripheral; Phenotype; Population; Quantitative Trait Loci; RNA analysis; Regenerative Medicine; Reproducibility; Research Personnel; Risk; Sampling; Severities; Single Nucleotide Polymorphism; Synapses; Time; Tissues; Transcriptional Regulation; Variant; Vial device; aged; autism spectrum disorder; behavioral phenotyping; biological systems; case control; chromatin remodeling; clinical phenotype; cost; de novo mutation; differential expression; disorder control; exome sequencing; gene discovery; genetic makeup; genomic locus; in vivo; individuals with autism spectrum disorder; induced pluripotent stem cell; inhibitory neuron; insight; live cell imaging; neurodevelopment; potential biomarker; relating to nervous system; risk variant; sex; social; stem cell genes; therapeutic target; transcriptome; transcriptome sequencing

Autism Spectrum Disorder (ASD) is a genetically and phenotypically heterogeneous neurodevelopmental disorder. Hundreds of genes contribute to the risk to develop ASD with no individual genetic locus accounting for more than 1% of cases. This raises the issue of whether, and how such diverse mechanisms converge on a smaller number of biological pathways that ultimately result in one phenotype, namely ASD. The inability to study neurons and brains from living subjects has blocked progress toward understanding the cellular and molecular mechanisms underlying ASD and other neurodevelopmental disorders. Neurons derived from human induced pluripotent stem cell (hiPSC) recapitulate multiple stages of in vivo neural development in vitro. Because they retain the genetic makeup of the patient they enable in vitro studies of neurons that harbor the complex genetic background associated with ASD. Deriving neurons from hiPSCs is labor intensive and costly, and to date studies have examined very small numbers of patients. hiPSC studies of the scope required to capture idiopathic ASD have not been possible, entirely limiting our ability to determine the mechanistic underpinnings of this form of the disorder. We propose to conduct an investigation of hiPSC derived neurons on an unprecedented scale by leveraging our California Institute for Regenerative Medicine (CIRM) funded existing collection of over 300 hiPSCs, to examine 100 individuals with idiopathic ASD and 100 age- and sex-matched controls. We will use commercially derived neurons (iCell Neurons) which are a >95% pure population of glutamatergic (excitatory) and GABAergic (inhibitory) neurons. To identify dysregulated molecular pathways in these neurons we will sequence the human transcriptome at three time points during maturation of the neurons. We will also conduct assays of cell viability, morphology, neurite outgrowth using live cell imaging, and function through calcium imaging. We aim to identify dysregulated molecular pathways in these hiPSC-derived neurons from individuals with ASD, by identifying genes that are differentially expressed from controls at each time point and perform a factorial analysis to study interactive effects between time point and disease variable. This will identify genes showing differentiation- induced expression changes across neuronal differentiation in individuals with ASD. We will identify key drivers of biological pathway changes using Weighted Gene Co-expression Network Analysis (WGCNA). Finally, we will relate gene expression profiles to cellular and behavioral phenotypes.

自闭症谱系障碍（ASD）是一种遗传和表型异质的 神经发育障碍。数百个基因有助于发展ASD的风险，而没有个体 遗传基因座占病例的1％以上。这就提出了是否以及如此多样化的问题 机制在较小数量的生物途径上收敛，最终导致一种表型， 即阿斯德。无法研究生活受试者的神经元和大脑的进展阻碍 了解ASD和其他神经发育的细胞和分子机制 疾病。 源自人类诱导多能干细胞（HIPSC）的神经元概括了多个阶段 体内神经发育体外。因为他们保留了患者的遗传构成，所以可以在体外启用 具有与ASD相关的复杂遗传背景的神经元的研究。衍生神经元 HIPSC是劳动力密集且昂贵的，迄今为止，研究检查了少数患者。 HIPSC对捕获特发性ASD所需范围的研究是不可能的，完全限制了我们的 确定这种疾病形式的机械基础的能力。我们建议进行 通过利用我们的加利福尼亚研究所，以前所未有的规模调查HIPSC衍生的神经元 再生医学（CIRM）资助了300多个HIPSC的现有收藏，以检查100名患有 特发性ASD以及100岁和性别匹配的对照。我们将使用商业派生的神经元（icell 神经元）是谷氨酸能（兴奋性）和GABA能（抑制性）神经元的纯纯度群体。 为了鉴定这些神经元中的分子途径失调，我们将对人类转录组进行测序 神经元成熟期间的三个时间点。我们还将进行细胞活力，形态，形态， 使用活细胞成像的神经突生长，并通过钙成像功能。我们旨在确定 这些HIPSC衍生的ASD个体中的分子途径失调，通过识别 从每个时间点从对照组中差异表达并进行阶乘分析以研究的基因 时间点和疾病变量之间的互动效应。这将确定显示出分化的基因 - 诱导ASD个体神经元分化的表达变化。我们将确定关键 使用加权基因共表达网络分析（WGCNA）变化生物途径的驱动因素。 最后，我们将将基因表达谱与细胞和行为表型联系起来。