Using genetics to uncover cellular phenotypes underlying Dup15q syndrome

染色体15 q11-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， 具有染色体15 q（Dup 15 q）重复的个体也患有通常共病的特征， 患有特发性自闭症--发育迟缓，运动技能迟缓，和癫痫。Dup 15 q综合征是一种完全- 由一个跨越~15-20个基因的区域的一个或两个额外拷贝的存在引起的渗透性疾病，但 负责的特定基因尚不清楚。基于人类遗传研究和小鼠模型，我们 假设UBE 3A加上至少一个其它基因表达增加导致表型 Dup 15 q综合征为了验证这一假设，我们将使用CRISPR和LoxP技术， 来源于Dup 15 q个体的遗传正确的人诱导多能干细胞（iPSC） 综合征然后，我们将对这些神经元进行详细的电生理表征。 同基因Dup 15 q/对照iPSC对，以鉴定与Dup 15 q综合征相关的细胞表型。 初步数据表明，Dup 15 q iPSC衍生的神经元是过度兴奋的，并且在突触连接中存在缺陷。 可塑性和稳态突触缩放。最后，我们将使用CRISPR抑制和反义寡核苷酸， 单独或组合地减少复制区域中基因表达的技术， 它们对人类Dup 15 q神经元的细胞病理生理学的贡献。成功完成这些 实验将确定Dup 15 q综合征的细胞表型以及导致Dup 15 q综合征的基因。 他们这里收集的信息将为药物发现创造细胞资源， 更好的Dup 15 q综合征小鼠模型，并确定可能有助于Dup 15 q综合征的神经元病理生理学。 综合征型和特发性自闭症。

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.