Using genetics to uncover cellular phenotypes underlying Dup15q syndrome

染色体15q11-q13的重复是最常见的染色体异常之一 自闭症。除了与自闭症相关的社交、言语/语言和重复行为缺陷外， 染色体15q(Dup15q)重复的个体也患有经常合并疾病的特征 患有特发性自闭症--发育迟缓、运动技能迟缓和癫痫发作。Dup15q综合征是一种完全- 由存在一个或两个额外拷贝的跨越约15-20个基因的区域引起的穿透性疾病，但 具体的基因(S)的责任还不清楚。基于人类基因研究和小鼠模型，我们 假设UBE3A的表达增加加上至少一个其他基因导致表型 Dup15q综合征的临床表现。为了验证这一假设，我们将使用CRISPR和loxP技术来 基因正确的人诱导多能干细胞(IPSCs)来源于携带Dup15q的个体 综合症。然后，我们将对由这些细胞产生的神经元进行详细的电生理表征 等位基因Dup15q/对照IPSC对用于鉴定与Dup15q综合征相关的细胞表型。 初步数据表明，Dup15q iPSC来源的神经元过度兴奋，突触存在缺陷 可塑性和稳态突触伸缩。最后，我们将使用CRISPR抑制和反义寡核苷酸 单独或组合减少复制区域中的基因表达以确定 它们对人Dup15q神经元细胞病理生理学的贡献。成功完成这些任务 实验将确定Dup15q综合征的细胞表型以及与Dup15q综合征相关的基因 他们。这里收集的信息将为药物发现创建细胞资源，通知构建 更好的Dup15q综合征小鼠模型，并确定可能有助于 自闭症的综合征和特发性形式。

Duplications of chromosome 15q11-q13 are one of the most common chromosomal anomalies associated with autism. In addition to the social, speech/language, and repetitive behavior deficits associated with autism, individuals with duplications of chromosome 15q (Dup15q) also suffer from features that are frequently co-morbid with idiopathic autism--developmental delay, motor skills delay, and seizures. Dup15q syndrome is a fully- penetrant disorder caused by the presence of one or two extra copies of a region spanning ~15-20 genes, but the specific gene(s) responsible are not clear. Based on human genetic studies and mouse models, we hypothesize that increased expression of UBE3A plus at least one other gene leads to the phenotypic manifestations of Dup15q syndrome. To test this hypothesis, we will use CRISPR and LoxP technologies to genetically correct human induced pluripotent stem cells (iPSCs) derived from individuals with Dup15q syndrome. We will then perform detailed electrophysiological characterization of neurons generated from these isogenic Dup15q/control iPSC pairs to identify cellular phenotypes associated with Dup15q syndrome. Preliminary data suggests that Dup15q iPSC-derived neurons are hyperexcitable and have deficits in synaptic plasticity and homeostatic synaptic scaling. Finally, we will use CRISPR inhibition and antisense oligonucleotide technologies to reduce expression of genes in the duplicated region individually or in combination to determine their contribution to the cellular pathophysiology in human Dup15q neurons. Successful completion of these experiments will identify cellular phenotypes underlying Dup15q syndrome as well as the genes contributing to them. The information garnered here will create cellular resources for drug discovery, inform construction of better mouse models for Dup15q syndrome, and identify neuronal pathophysiology that may contribute to syndromic and idiopathic forms of autism.