Synaptic signaling in a human stem cell model of Angelman syndrome

天使综合征人类干细胞模型中的突触信号传导

基本信息

批准号：
8292528
负责人：
Eric S Levine
金额：
$ 19.25万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
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：
2012-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8292528
关键词：
Action Potentials Address Affect Age Alleles Angelman Syndrome Animal Model Autistic Disorder Basic Science Behavioral Brain Calcium/calmodulin-dependent protein kinase Cell Culture Techniques Cell Line Cell model Cells Chromosome Deletion Complex Defect Dendritic Spines Dermal Development Disease Excitatory Synapse Fibroblasts Genes Genetic Genetically Engineered Mouse Genomics Genotype Human Immediate-Early Genes Impaired cognition Impairment In Vitro Individual Intellectual functioning disability Language Measures Mediating Modeling Molecular Motor Ataxias Movement Disorders Neurons Patients Pattern Phenotype Physiological Play Property Regulation Research Role Sampling Screening procedure Seizures Signal Transduction Somatic Cell Speech Synapses Synaptic Transmission Synaptic plasticity Syndrome Testing Therapeutic Tissues Ubiquitin-Protein Ligase Complexes base calmodulin-dependent protein kinase II density developmental disease disease phenotype drug discovery ephexin human disease human stem cells human tissue imprint induced pluripotent stem cell loss of function neurogenetics neuronal excitability new therapeutic target novel receptor relating to nervous system research study response synaptic function ubiquitin-protein ligase voltage

项目摘要

DESCRIPTION (provided by applicant): Individuals with a deletion of chromosome 15q11-q13 suffer from Angelman syndrome (AS), a neurogenetic developmental disorder characterized by intellectual disability, motor ataxia, absent speech, and seizures. The specific gene that is responsible for AS encodes the ubiquitin protein ligase UBE3A. In AS, deficits in synaptic signaling and plasticity appear to play a critical role in the disease phenotype, but the exact functional role of UBE3A and its relevant downstream targets are unknown. In order to develop appropriate treatments for AS, it is necessary to understand the pathophysiological changes caused by UBE3A deletion. Until recently it has not been possible to examine the functional properties of brain neurons in affected individuals. The discovery of genomic reprogramming of human somatic cells into induced pluripotent stem cell (iPSC) lines provides a novel way to model human diseases with complex genetics. We have recently succeeded in reprogramming dermal fibroblasts from AS patients, as well as age-matched control subjects, into iPSCs, and then differentiated these cells into functional neurons that maintain the imprinting phenotype of UBE3A expression seen in AS patients. We are now poised to take advantage of these novel patient-derived cell lines to test specific hypotheses about the underlying physiological defects in AS. The first aim uses electrophysiological and immunocytochemical approaches to explore the intrinsic functional properties of iPSC-derived neurons and activity-dependent plasticity of synaptic connections. Rescue experiments will focus on the roles of the immediate early gene ARC and the calcium/calmodulin-dependent protein kinase CaMKII in AS-associated deficits in synaptic signaling. The second aim will explore changes in synapse number and dendritic spine density in AS-derived neurons. Rescue experiments will target the role of ephexin-5, a substrate of UBE3A that plays a role in regulating excitatory synapse number during development. Overall, this approach may prove useful for identifying novel targets for drug discovery and for screening potential therapeutics aimed at ameliorating and/or curing the seizures, movement disorders, and language and cognitive impairments in Angelman syndrome. PUBLIC HEALTH RELEVANCE: Angelman syndrome is a neurogenetic developmental disorder characterized by intellectual disability, motor ataxia, absent speech, and seizures. The proposed research uses a novel human stem cell culture model to investigate the cellular and molecular basis of these deficits. The long term objective is to identify novel therapeutic targets for treating Angelman syndrome and other autism-related disorders.

描述（由申请人提供）：具有15q11-Q13染色体染色体的个体患有Angelman综合征（AS），这是一种以智力残疾，运动性共济失调，缺乏言语和癫痫发作为特征的神经遗传发育障碍。特定基因是负责编码泛素蛋白连接酶UBE3A。在AS中，突触信号和可塑性的缺陷似乎在疾病表型中起着关键作用，但是UBE3A及其相关下游靶标的确切功能作用尚不清楚。为了为AS开发适当的治疗方法，有必要了解UBE3A缺失引起的病理生理变化。直到最近，还无法检查受影响个体中脑神经元的功能特性。将人体细胞的基因组重编程到诱导的多能干细胞（IPSC）系的发现提供了一种新颖的方法，可以模拟具有复杂遗传学的人类疾病。我们最近成功地从患者和年龄匹配的对照组中重新编程了皮肤成纤维细胞，然后将这些细胞区分为功能性神经元维持在患者中看到的UBE3A表达的烙印表型。现在，我们准备利用这些新型患者衍生的细胞系来测试有关AS中潜在的生理缺陷的特定假设。第一个目的使用电生理和免疫细胞化学方法来探索IPSC衍生神经元的内在功能特性以及突触连接的活性依赖性可塑性。救援实验将重点关注直接的早期基因弧和钙/钙调蛋白依赖性蛋白激酶CaMKII在突触信号中与AS相关缺陷中的作用。第二个目标将探索AS衍生神经元中突触数和树突状脊柱密度的变化。救援实验将瞄准ephexin-5的作用，ephexin-5是UBE3A的底物，在调节发育过程中起作用起作用。总体而言，这种方法可能证明可用于确定旨在改善和/或治愈Angelman综合征中的癫痫发作，运动障碍以及语言和认知障碍的潜在治疗剂的新型靶标。公共卫生相关性：Angelman综合征是一种神经遗传发育障碍，其特征是智力残疾，共济失调，缺乏言语和癫痫发作。拟议的研究使用一种新型的人类干细胞培养模型来研究这些缺陷的细胞和分子基础。长期目标是确定新颖的治疗靶标用于治疗Angelman综合征和其他与自闭症有关的疾病。