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

基本信息

批准号：
8537524
负责人：
MATTHEW P ANDERSON
金额：
$ 36.73万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8537524
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 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 RNA Sequences 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 cell type chromatin immunoprecipitation cofactor design disability dosage gamma-Aminobutyric Acid genome wide association study insight mRNA Expression mouse model nervous system disorder neurobiological mechanism novel promoter protein degradation reconstitution response selective expression synaptic function therapeutic target therapy development tool trait ubiquitin ligase ubiquitin-protein ligase

项目摘要

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）：该项目的长期目标是比较和对比由Ube3a基因剂量改变的神经元mRNA表达和电路功能的缺陷，这些缺陷可能是自闭症谱系障碍（ASD）与Maternal 15Q11-11-13 Replication和Angelman的综合征（ASD）的行为表型的基础。 15q11-13 ASD和患者的细胞分别具有过量和缺乏的E3泛素连接酶Ube3a。 ube3a是从大脑中母体等位基因选择性表达的唯一基因。从母亲那里继承15q11-13重复的大多数儿童会发展为ASD，而遗传的副本越多，损害就越严重。最近，我们确定增加了UBE3A基因剂量仅重构小鼠自闭症相关的行为缺陷的三合会。相比之下，当儿童继承母体UBE3A基因突变时，Angelman's综合征（AS）会产生。重要的是，UBE3A还充当了与连接酶活性无关的基因表达调节基因表达的核受体共激活器。因此，我们假设过量的UBE3A（15Q DUP模型）起作用在细胞核中，以扰动特定神经元基因和编码蛋白的正常表达以改变电路功能，从而产生与自闭症相关的行为缺陷。逻辑上的推论是，核UBE3A的缺陷可能会改变基因表达以改变电路功能，并通过相同但相反的机制产生与Angelman相关的行为缺陷。由于大脑的电路设计用于定量评估感觉信息和定量运动响应，因此当数量改变数量时，许多控制电路功能的分子可能会导致行为的分级变化。我们提出UBE3A通过对神经元基因表达的剂量依赖性调节来做到这一点。这种分子量变化可能是ASD等行为障碍的内在异质性的基础。此R01赠款的具体目的如下：（1和2）确定UBE3A基因剂量是否导致ASD（过量的UBE3A）和AS（缺乏UBE3A）通过其在细胞核内的作用来实现相关行为缺陷；（3）确定由UBE3A过量或缺乏定量调控的皮质神经元mRNA；（4）确定由UBE3A的过量和缺乏诱导的皮质电路功能障碍。我们的方法包括一种新型的核定核3a转基因，一种神经元细胞类型的UBE3A的特异性表达，转录本的基因组广泛分析以及鉴定UBE3A-GENE启动子结合位点的鉴定。切片电生理学和神经元形态将检查全球和细胞类型的特异性变化以及UBE3A基因剂量的核靶向变化对活脑切片中皮质回路功能的影响。该分析将促进我们的敌方从基因到电路到行为的桥梁，并在两种对比的人类神经疾病中。该项目促进了该机构的使命，以进一步深入了解ASD和通过遗传模型涉及的神经元细胞，电路和基因。新颖的分子见解和遗传工具将促进这些终身行为的治疗残疾。