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性的疾病 神经发育障碍。数百个基因会导致患自闭症谱系障碍（ASD）的风险，而没有个体的参与 遗传位点占病例的1%以上。这就提出了这样的问题：是否以及如何实现这种多样化 机制集中在较少数量的生物途径上，最终导致一种表型， 即自闭症谱系障碍（ASD）。无法研究活体受试者的神经元和大脑，阻碍了这方面的进展 了解 ASD 和其他神经发育的细胞和分子机制 失调。 源自人类诱导多能干细胞 (hiPSC) 的神经元重现了多个阶段 体内神经发育体外。因为它们保留了患者的基因组成，因此可以在体外进行 对具有与自闭症谱系障碍相关的复杂遗传背景的神经元的研究。从中导出神经元 hiPSC 是劳动密集型且昂贵的，迄今为止的研究只检查了极少数患者。 hiPSC 对捕获特发性 ASD 所需范围的研究尚未实现，这完全限制了我们的研究 确定这种疾病形式的机制基础的能力。我们建议开展一次 利用我们的加州研究所对 hiPSC 衍生神经元进行前所未有的研究 再生医学 (CIRM) 资助了现有 300 多个 hiPSC 的收集，以检查 100 名患有 特发性 ASD 和 100 名年龄和性别匹配的对照。我们将使用商业衍生的神经元（iCell 神经元），其纯度>95%，由谷氨酸能（兴奋性）和 GABA 能（抑制性）神经元组成。 为了识别这些神经元中失调的分子途径，我们将对人类转录组进行测序 神经元成熟过程中的三个时间点。我们还将进行细胞活力、形态学、 使用活细胞成像观察神经突的生长，并通过钙成像观察其功能。我们的目标是确定 通过鉴定来自 ASD 个体的 hiPSC 衍生神经元中失调的分子通路 在每个时间点与对照差异表达的基因并进行因子分析来研究 时间点和疾病变量之间的交互作用。这将识别显示分化的基因- 诱导自闭症谱系障碍 (ASD) 个体神经元分化过程中的表达变化。我们将确定关键 使用加权基因共表达网络分析 (WGCNA) 确定生物途径变化的驱动因素。 最后，我们将基因表达谱与细胞和行为表型联系起来。