Neurobiological Mechanism of 15q11-13 Duplication Autism Spectrum Disorder

15q11-13重复自闭症谱系障碍的神经生物学机制

• 批准号: 9128069
• 负责人: MATTHEW P ANDERSON
• 金额: $ 38.06万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9128069
• 关键词: 15q; Affect; Alleles; Angelman Syndrome; Antibodies; Ataxia; Autistic Disorder; Behavior; Behavior Disorders; Behavioral; Binding Sites; Brain; Cell Line; Cell Nucleus; Cells; Central Nervous System Diseases; Cerebellum; Cerebral cortex; ChIP-seq; Child; Chromosomes, Human, Pair 15; Cognitive deficits; Communication; Consensus; Copy Number Polymorphism; Defect; Disease; Dose; Electrophysiology (science); Epilepsy; Epitopes; Estrogen Receptors; Functional disorder; Gene Dosage; Gene Expression; Gene Mutation; Genes; Genetic; Genetic Models; Genotype; Glucocorticoid Receptor; Glutamates; Grant; Heterogeneity; Human; Immunofluorescence Immunologic; Impairment; In Situ; In Vitro; Inherited; Isodicentric Chromosome; Knock-out; Learning; Length; Life; Ligase; Messenger RNA; Methods; Mission; Modeling; Molecular; Morphology; Mothers; Motor; Mus; Mutation; Neurobiology; Neuroglia; Neurons; Nuclear; Nuclear Export; Nuclear Receptors; Patients; Personality; Phenotype; Proteins; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Role; Sensory; Signal Transduction; Slice; Social Interaction; Substrate Specificity; Synapses; Testing; Transcript; Transgenes; Transgenic Mice; Transgenic Organisms; Triad Acrylic Resin; Ubiquitin; Variant; autism spectrum disorder; brain circuitry; cell type; cofactor; design; disability; dosage; gamma-Aminobutyric Acid; genome-wide analysis; insight; mRNA Expression; mouse model; nervous system disorder; neurobiological mechanism; novel; overexpression; promoter; protein degradation; reconstitution; repetitive behavior; response; selective expression; synaptic function; therapeutic target; therapy development; tool; trait; transcriptome sequencing; ubiquitin ligase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): The long-term objective of this project is to compare and contrast the defects in neuronal mRNA expression and circuit functions produced by altered Ube3a gene dosage that may underlie the behavioral phenotypes of Autism Spectrum Disorder (ASD) with maternal 15q11-13 duplication and Angelman's Syndrome (AS). Cells from 15q11-13 ASD and AS patients have excess and deficient E3 ubiquitin ligase, Ube3a, respectively. Ube3a is the sole gene expressed selectively from the maternal allele in brain. The majority of children who inherit 15q11-13 duplications from their mothers develop ASD and the more copies inherited, the more severe the impairment. Recently, we established that increasing Ube3a gene dosage alone reconstitutes the triad of autism-related behavioral deficits in mice. Angelman's Syndrome (AS), by contrast, results when children inherit maternal Ube3a gene mutations. Importantly, Ube3a also acts as a nuclear receptor co-activator regulating gene expression independent of ligase activity. Thus we hypothesized that excess Ube3a (15q dup model) acts in the nucleus to perturb the normal expression of specific neuronal genes and encoded proteins to alter circuit function and thereby generate autism-related behavioral deficits. The logical corollary is that deficiencies of nuclear Ube3a may alter gene expression to change circuit function and produce Angelman-related behavioral deficits through the same, but opposite mechanisms. Because the brain's circuitries are designed for quantitative assessment of sensory information and quantitative motor responses, it is likely that many molecules controlling circuit functions can cause graded changes in behavior when their quantities are altered. We propose Ube3a do so through a dose-dependent regulation of neuronal gene expression. Such molecular quantity variations likely underlie the intrinsic heterogeneity of behavioral disorders like ASD. The specific aims of this R01 grant follow: (1 and 2) determine if Ube3a gene dosage causes ASD (excess Ube3a) and AS (deficient Ube3a) related behavioral deficits through its actions within the nucleus; (3) identify the cortical neuron mRNAs quantitatively regulated by excesses or deficiencies of Ube3a; and (4) identify the cortical circui dysfunctions quantitatively induced by excesses and deficiencies of Ube3a. Our methods include a novel nuclear-targeted Ube3a transgene, a neuron cell-type specific expression of Ube3a, genome wide analysis of transcripts, and identification of consensus Ube3a-gene promoter binding sites. Slice electrophysiology and neuronal morphology will examine the effects of global and cell-type specific changes and nuclear targeted changes of Ube3a gene dosage on cortical circuit function in living brain slices. This analysis will facilitate our effors to bridge from genes to circuits to behavior in the two contrasting human neurological diseases. The project promotes the agency's mission to further a deeper understanding of the neuronal cells, circuits, and genes involved in ASD and AS via genetic models. The novel molecular insights and genetic tools will facilitate development of treatments for these life-long behavioral disabilities.

描述（由申请人提供）：本项目的长期目标是比较和对比Ube3a基因剂量改变所产生的神经元mRNA表达和电路功能的缺陷，这些缺陷可能是具有母体15q11-13重复的自闭症谱系障碍（ASD）和Angelman综合征（AS）行为表型的基础。来自15q11-13 ASD和AS患者的细胞分别存在E3泛素连接酶Ube3a过量和缺乏。Ube3a是脑内唯一由母体等位基因选择性表达的基因。大多数从母亲那里继承了15q11-13重复基因的孩子会患上自闭症，而且遗传的副本越多，损害就越严重。最近，我们发现单独增加Ube3a基因剂量可以重建小鼠自闭症相关行为缺陷的三联征。相比之下，安吉尔曼综合征（AS）是由于儿童继承了母亲的Ube3a基因突变而导致的。重要的是，Ube3a还作为核受体共激活因子，独立于连接酶活性调节基因表达。因此，我们假设过量的Ube3a （15q dup模型）在细胞核中作用，扰乱特定神经元基因和编码蛋白的正常表达，从而改变回路功能，从而产生自闭症相关的行为缺陷。合乎逻辑的推论是，细胞核Ube3a缺陷可能通过相同但相反的机制改变基因表达，从而改变回路功能，并产生与天使相关的行为缺陷。因为大脑回路是为定量评估感觉信息和定量运动反应而设计的，所以很可能许多控制回路功能的分子在数量改变时，会导致行为的渐变变化。我们建议Ube3a通过对神经元基因表达的剂量依赖性调节来实现这一目标。这种分子数量的变化可能是ASD等行为障碍的内在异质性的基础。该R01拨款的具体目的如下：(1)和(2)确定Ube3a基因剂量是否通过其在细胞核内的作用导致ASD（过量Ube3a）和AS（缺乏Ube3a）相关的行为缺陷；(3)定量鉴定受Ube3a过量或缺乏调控的皮质神经元mrna；(4)定量鉴定由Ube3a过量和缺乏引起的皮质回路功能障碍。我们的方法包括一种新的核靶向Ube3a转基因，Ube3a的神经元型特异性表达，转录本的全基因组分析，以及Ube3a基因启动子结合位点的鉴定。切片电生理学和神经元形态学将检测Ube3a基因剂量对活脑切片皮层回路功能的整体和细胞类型特异性变化以及核靶向性变化。这一分析将有助于我们在两种截然不同的人类神经疾病中从基因到电路再到行为的桥梁。该项目促进了该机构的使命，即通过遗传模型进一步深入了解与ASD和AS有关的神经细胞、电路和基因。新的分子见解和遗传工具将促进治疗这些终身行为的发展