High Content Screens of Neuronal Development for Autism Research

描述（由申请人提供）：神经元回路的异常发育被认为是自闭症谱系障碍（ASD）和相关疾病（如Rett综合征）的特征性行为表现的基础。在ASD中已经记录的特定脑区域的异常大小和功能可能是由神经元和突触的异常发育引起的，这些特征可以使用体外方法进行量化。本申请提出开发一种廉价的、基于细胞的神经元形态发生和突触发生的测定，其将促进ASD的基础研究和临床前药物开发。基于形态学的测定将适合于对来自动物模型或来自患有ASD和相关疾病的人类患者的诱导多能干细胞（iPS细胞）的神经元进行自动化、高内容筛选。将对神经突生长、树突分支、棘形状和突触标记物进行定量。将进行概念验证实验以评价与未受影响的个体相比，来自Rett综合征患者的iPS细胞（由合作者提供）中神经元发育的差异。 公共卫生相关性：该试点项目将开发定量的、基于显微镜的神经元发育分析，可用于评估自闭症和相关疾病患者的诱导多能干细胞系衍生的神经元。该测定将有助于a）评估患者群体中和患有自闭症综合征的个体中神经元发育的异质性; B）研究异常神经元发育的分子机制;和c）筛选可改善自闭症中正常神经元发育的药物化合物。

DESCRIPTION (provided by applicant): Abnormal development of neuronal circuitry is thought to underlie the characteristic behavioral manifestations of autism spectrum disorder (ASD) and related disorders like Rett syndrome. The aberrant size and function of specific brain regions that has been documented in ASD is likely to result from aberrant development of neurons and synapses, features that can be quantified using in vitro approaches. This application proposes to develop an inexpensive, cell-based assay of neuronal morphogenesis and synaptogenesis that will facilitate both basic research and pre-clinical drug development for ASD. The morphology-based assay will be amenable to automated, high-content screening of neurons derived from either animal models or from induced pluripotent stem cells (iPS cells) from human patients with ASD and related disorders. Neurite outgrowth, dendritic branching, spine shape, and synaptic markers will be quantified. Proof of concept experiments will be conducted to evaluate differences in neuronal development in iPS cells (supplied by collaborator) from Rett syndrome patients compared to unaffected individuals. PUBLIC HEALTH RELEVANCE: This pilot project will develop quantitative, microscopy-based assays of neuronal development that can be used to evaluate neurons derived from lines of induced pluripotent stem cells of patients with autism and related disorders. The assay will facilitate a) the assessment of heterogeneity in neuronal development among patient populations and across individuals with autistic syndromes; b) investigation of molecular mechanisms underlying abnormal neuronal development; and c) screening of pharmaceutical compounds that may improve normal neuronal development in autism.