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）

基本信息

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
项目类别：
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

项目摘要

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（由适用提供）：本研究旨在生成强大的工具和工作流程，以创建基于自闭症谱系障碍（ASD）的基于人类的多能干细胞（HIPSC）模型，并开发可扩展的分子和细胞表型相关的可扩展测定法。我们已经确定了hipsC宽度采用的几个关键瓶颈是允许神经疾病基础的分子机制并启用临床前药物筛查的工具。我们组建了一个由五个神经科学，干细胞生物学和计算生物学领域的主要专家组成的团队，他们将与三家创新驱动的生物技术公司（Fluidigm，BD Biosciences and Synthetic Genomics）合作，以克服这些障碍。由于自闭症被认为是突触发育和功能障碍，最终导致大脑的电路功能障碍，因此我们将开发出突触终端网络功能的定量测定法，可用于高通量药物筛查。我们还旨在揭示宝贵的合成和网络失调的上游分子事件，并确定预测性的RNA和蛋白质特征。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，使用特征良好的神经型HIPSC系列作为起点。然后，我们将把这些正常细胞和突变细胞与皮质神经元和星形胶质细胞区分开，这两种细胞类型最强烈与自闭症病理生理学有关。在单细胞水平上高度定量和敏感的测定将用于鉴定可以区分ASD神经元和星形胶质细胞与正常细胞的蛋白质和RNA表达的变化。最后，我们将使用可以以高通量格式使用的先进技术来制定测量突触密度和强度的测定法。我们设想我们的工具，技术和测定法，所有这些都将在生成时公开可用，这两个都将有助于我们对ASD和加速神经疾病的临床前研究的理解。