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

临床前自闭症细胞检测、生物特征和网络分析方面的合作（Copacabana）

• 批准号: 8935692
• 负责人: Eugene Wei-Ming Yeo
• 金额: $ 280.66万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-21 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8935692
• 关键词: Adoption; Astrocytes; Autistic Disorder; Biological Assay; Biology; Brain; Brain Diseases; Cell Line; Cell Therapy; Cell model; Cells; Cellular Assay; Collaborations; Complex; Computational Biology; Data Set; Development; Diagnostic; Diagnostic Procedure; Disease; Disease model; Dissection; Drug Targeting; Engineering; Event; Functional disorder; Genetic; Genetic Engineering; Genetic Models; Genetic Transcription; Genome engineering; Genomics; Haplotypes; Helper Viruses; Heterogeneity; Human; Image Analysis; In Vitro; Individual; Institutes; Label; Laboratories; Measures; Medicine; Mental Health; Methods; Modeling; Molecular; Molecular Profiling; Mus; Neurites; Neuroglia; Neurons; Neurosciences; Normal Cell; Patients; Phenotype; Preclinical Drug Evaluation; Process; Proteins; Protocols documentation; RNA; Rabies virus; Research; Resources; Stem cells; Symptoms; Synapses; Technology; Testing; Variant; autism spectrum disorder; base; biosignature; cell type; density; drug discovery; genetic variant; genome editing; imaging modality; improved; induced pluripotent stem cell; inhibitory neuron; innovation; insight; mutant; nervous system disorder; neural circuit; novel; novel diagnostics; pre-clinical; pre-clinical research; protein expression; public health relevance; public-private partnership; screening; stem cell biology; synaptogenesis; tool

 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.

 描述（由申请人提供）：本研究旨在生成用于创建基于人诱导多能干细胞（hIPSC）的自闭症谱系障碍（ASD）模型的强大工具和工作流程，并开发用于预测自闭症相关分子和细胞表型的可扩展测定。我们已经确定了广泛采用hIPSC作为工具的几个关键瓶颈，这些工具允许解剖神经系统疾病的分子机制并实现临床前药物筛选。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Synthetic Genomics）合作，以克服这些障碍。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，我们将开发突触末端网络功能的定量分析，可用于高通量药物筛选。我们的目标还在于揭示引发突触和网络失调的上游分子事件，并识别预测性RNA和蛋白质特征。我们的策略是使用良好表征的神经典型hIPSC系作为起点，通过基因组操作来工程化自闭症遗传形式的模型。然后，我们将这些正常和突变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病理生理学关系最密切。在单细胞水平上的高度定量和灵敏的测定将用于鉴定蛋白质和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。最后，我们将开发使用先进技术测量突触密度和强度的测定方法，这些技术可以用于高通量格式。我们设想，我们的工具，技术和检测方法，所有这些我们将在生成时公开提供，将为我们对ASD的理解做出重要贡献，并加速神经系统疾病的临床前研究。