Neurobiology of Autism With Macrocephaly

自闭症大头畸形的神经生物学

• 批准号: 9920231
• 负责人: FLORA M VACCARINO
• 金额: $ 46.24万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-22 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920231
• 关键词: Affect; Binding Sites; Biological; Biological Assay; Biological Process; Brain; Cell Adhesion; Cell Line; Cell Proliferation; Cell division; Cells; ChIP-seq; Child; Chromatin; Code; Collection; DNA Sequence Alteration; Data; Data Set; Development; Electrophysiology (science); Epigenetic Process; Etiology; FOXG1B gene; Family; First Pregnancy Trimester; Forebrain Development; Future; Gene Expression; Genes; Genetic; Genetic Heterogeneity; Genetic Transcription; Genome; Genomic Segment; Genotype; Glutamates; Goals; Growth; Heterogeneity; Histones; Human; In Vitro; Individual; Inherited; Macrocephaly; Maps; Measures; Neurobiology; Neuronal Differentiation; Neurons; Nucleic Acid Regulatory Sequences; Organoids; Pathogenesis; Pathway Analysis; Pathway interactions; Patients; Phenotype; Proteins; RNA Interference; Regulator Genes; Ribonucleic Acid Regulatory Sequences; Role; Source; Structure; Subgroup; Synapses; Technology; Testing; Therapeutic; Untranslated RNA; Validation; Variant; autism spectrum disorder; brain size; brain volume; differential expression; experimental study; gamma-Aminobutyric Acid; genetic variant; genome sequencing; genome-wide; genomic variation; induced pluripotent stem cell; inhibitory neuron; insight; knock-down; molecular marker; nerve stem cell; neurodevelopment; outcome forecast; overexpression; patient subsets; proband; progenitor; programs; public health relevance; rare variant; structural genomics; synaptogenesis; trait; transcription factor; transcriptome; transcriptome sequencing; whole genome

 DESCRIPTION (provided by applicant): Autism spectrum disorders (ASDs) affect 1%-2.5% of children worldwide. We suggest that etiological and genetic heterogeneity might converge in a few neurobiological downstream pathways. We have been investigating the pathobiology of ASD with large brain volume (macrocephaly), a phenotype which confers poorer prognosis. Ongoing studies have shown that telencephalic organoids differentiated in vitro from induced pluripotent stem cells (iPSC) derived from patients with ASD and macrocephaly have increased cell proliferation, increased synaptic growth and overproduction of GABAergic inhibitory neurons, indicating an early imbalance in glutamate/GABA neuron ratio. RNA interference experiments suggested that the overproduction of GABAergic cells is attributable, at least in part, to an increase in expression of FOXG1, a master regulatory transcription factor crucial for telencephalic development. Major goals of this application are (1) to expand our analysis of the developmental pathways that are dysregulated in ASD to a larger number of families and (2) to understand to what extent developmental alterations we identified in ASD with macrocephaly also apply to ASD in general. To this end, we will obtain data on neurobiological measures, transcriptome and chromatin active regions in organoids derived from ASD patients with enlarged brain size and ASD patients with normal brain size. The altered gene regulatory network will be inferred and the two networks will be compared to understand similarities and differences in the two subgroups of ASD. To begin to understand the upstream causes of these developmental alterations, we will then investigate whether patients with ASD carry an increased burden of rare genomic variations in regions of the genome that participate in this regulatory network. Finally we will perform overexpression and RNAi knockdown experiments to examine the specific role of our current best candidate transcription factor, FOXG1, in the constellation of neurobiological and transcriptome alterations found in ASD-derived progenitors. We will assess the impact of perturbing FOXG1 gene expression on neurobiological functions (cell proliferation, glutamate/GABA neuron fate, synaptic growth), transcriptome and activity of transcription regulatory regions by RNA-seq and ChIP-seq, respectively, to gain insights into the role of a FOXG1-driven transcriptional program in the aberrant neuronal differentiation of ASD-derived neural progenitors. In summary, in this application we delineate strategies for (1) identifying gene networks and biological pathways that characterize altered development in two subgroups of ASD; (2) testing the causal role of one crucial node in such networks, the transcription factor FOXG1, which is over- active in ASD with macrocephaly; and (3) identifying regulatory factors, both genetic and epigenetic, upstream from neurobiological and gene expression abnormalities. The impact of these experiments will be the definition of a number of biological functions and molecular markers that are implicated in the neurobiology of ASD.

 描述（由适用提供）：自闭症谱系障碍（ASDS）影响全球1％-2.5％的儿童。我们认为，病因和遗传异质性可能会在一些神经生物学下游途径中汇聚。我们一直在研究ASD的病理生物学，其大脑体积较大（脑头畸形）是一种表现型，供认较差的预后。正在进行的研究表明，从ASD和脑头畸形患者中得出的诱导多能干细胞（IPSC）分化的尾类器官会增加细胞增殖，突触增长的增加和GABA能抑制性神经元的过量产生，表明glutamate/Gaba neuron heuron heuron heuron havaio rance baba rutagragic抑制性神经元。 RNA干扰实验表明，GABA能细胞的生产过多至少部分地归因于FOXG1表达的增加，FoxG1的表达是对脑脑发育至关重要的主要调节转录因子。该应用程序的主要目标是（1）扩展我们对ASD中失调的发育途径的分析到较大的家庭，以及（2）了解我们在ASD中与宏观畸形在ASD中确定的发展程度在多大程度上也适用于ASD。为此，我们将获得有关来自脑大小增加和正常脑大小的ASD患者的ASD患者的类器官中神经生物学措施，转录组和染色质活性区域的数据。将推断出改变的基因调节网络，并将两个网络进行比较，以了解ASD两个亚组的相似性和差异。为了开始了解这些发育改变的上游原因，我们将研究ASD患者是否会增加参与该调节网络的基因组区域的罕见基因组变异。最后，我们将进行过表达和RNAi敲低实验，以检查我们当前最佳候选转录因子FOXG1在ASD衍生祖细胞中发现的神经生物学和转录组改变的星座中的特定作用。我们将评估扰动FOXG1基因表达对神经生物学功能的影响（细胞增殖，谷氨酸/GABA神经元命运，突触生长），通过RNA-SEQ和CHIP-SEQ分别通过RNA-SEQ和CHIP-SEQ进行转录调节区域的活性，以使foxg1驱动的NERONS的作用分别获得依从型foxg1的作用。神经元祖细胞。总而言之，在此应用中，我们描述了（1）识别基因网络和生物学途径的策略，这些基因网络和生物学途径表征了ASD两个亚组中发展的变化； （2）测试一个关键节点在此类网络中的因果作用，即转录因子FOXG1，该因子在ASD中过度活跃； （3）从神经生物学和基因表达异常上游鉴定遗传和表观遗传学的调节因素。这些实验的影响将是在ASD神经生物学中实现的许多生物学功能和分子标记的定义。