Collaboration on preclinical autism cellular assays, biosignatures, and network analyses (Copacabana)

 描述(由申请人提供)：这项研究旨在生成强大的工具和工作流程，以创建基于人类诱导多能干细胞(HiPSC)的自闭症谱系障碍(ASD)模型，并开发可扩展的与自闭症相关的预测分子和细胞表型的分析方法。我们已经确定了广泛采用HiPSCs作为工具的几个关键瓶颈，这些工具允许剖析神经疾病的潜在分子机制，并使临床前药物筛选成为可能。我们已经组建了一个由神经科学、干细胞生物学和计算生物学领域的五位顶尖专家组成的团队，他们将与三家创新驱动的生物技术公司(Fluidigm、BD Biosciences和合成基因组学)合作克服这些障碍。由于自闭症被认为是一种突触发育和功能障碍，最终导致大脑回路功能障碍，我们将开发可用于高通量药物筛选的突触末端网络功能的定量分析。我们还旨在揭示导致突触和网络失调的上游分子事件，并识别预测性RNA和蛋白质信号。我们的策略是通过基因组操作来设计自闭症遗传形式的模型，使用具有良好特征的、神经典型的HiPSC系作为起点。然后，我们将把这些正常和突变的细胞分化为皮质神经元和星形胶质细胞，这两种细胞类型与自闭症的病理生理学关系最密切。单细胞水平的高定量和高灵敏度分析将用于识别蛋白质和RNA表达的变化，这些变化可以将ASD神经元和星形胶质细胞与正常细胞区分开来。最后，我们将开发使用先进技术测量突触密度和强度的分析方法，这些技术可以用于高通量格式。我们设想，我们的工具、技术和分析，所有这些我们将在产生时公开提供，都将对我们对ASD的理解做出关键贡献，并加快神经疾病的临床前研究。

 DESCRIPTION (provided by applicant): This study aims to generate robust tools and workflows for creating human induced pluripotent stem cell (hIPSC)-based models of autism spectrum disorder (ASD), and to develop scalable assays for predictive molecular and cellular phenotypes relevant to autism. We have identified several key bottlenecks in the widespread adoption of hIPSCs as tools that allow the dissection of molecular mechanisms underlying neurological disease and enable preclinical drug screening. We have assembled a team of five leading experts in neuroscience, stem cell biology and computational biology, who will collaborate up with three innovation-driven biotech companies (Fluidigm, BD Biosciences and Synthetic Genomics) to overcome these roadblocks. Since autism is considered a disorder of synapse development and function that ultimately leads to circuit dysfunction in the brain, we will develop quantitative assays of synapse end network function that can be used in high-throughput drug screens. We also aim to uncover the upstream molecular events that precipitate synaptic and network dysregulation, and identify predictive RNA and protein signatures. Our strategy is to engineer models of genetic forms of autism by genomic manipulation using a well- characterized, neurotypical hIPSC line as the starting point. We will then differentiate these normal and mutant cells to cortical neurons and astrocytes, the two cell types that have been most strongly implicated in autism pathophysiology. Highly quantitative and sensitive assays at the single-cell level will be used to identify changes in protein and RNA expression that can distinguish ASD neurons and astrocytes from normal cells. Finally, we will develop assays measuring synapse density and strength using advanced technology that can be used in high-throughput format. We envision that our tools, technologies and assays, all of which we will make publicly available as they are being generated, will both critically contribute to our understanding of ASD and accelerate preclinical research of neurological disease.