Improved Scalability of Neuron-Based Phenotypic Screening Assays for Therapeutic Discovery in Neuropsychiatric Disorders.
Improved Scalability of Neuron-Based Phenotypic Screening Assays for Therapeutic Discovery in Neuropsychiatric Disorders.
复制标题
提高神经精神疾病治疗发现的基于神经元的表型筛选测定的可扩展性。
DOI：
10.1159/000481731
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Davis,Ro
中科院分区：
文献类型：
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作者：
Spicer,TimothyP;Hubbs,Christopher;Vaissiere,Thomas;Collia,Deanna;Rojas,Camilo;Kilinc,Murat;Vick,Kyle;Madoux,Franck;Baillargeon,Pierre;Shumate,Justin;Martemyanov,KirillA;Page,DamonT;Puthanveettil,Sathya;Hodder,Peter;Davis,Ro
有一个迫切的需要，以改善有关的神经精神疾病（NSD）的药物发现的方法。神经精神疾病的治疗发现将受益于筛选分析，可以测量与疾病机制相关的复杂表型的变化。然而，跟踪复杂神经元表型（例如神经元连接）的传统测定显示出较差的可扩展性，并且与高通量筛选（HTS）程序不兼容。因此，我们创建了一个神经元表型测定平台，专注于提高能够跟踪疾病相关表型的基于神经元的测定的可扩展性和可负担性。首先，使用廉价的实验室级自动化，我们将原代神经元培养生产工业化，这使得能够在功能神经网络中创建可扩展的检测。然后，我们开发了一组表型测定，其基于从表达HTS相容报告基因的转基因小鼠培养原代神经元，所述HTS相容报告基因捕获疾病相关表型。我们证明，在典型的学术实验室环境中，仅使用几窝小鼠就可以快速筛选出1，280种化合物的库。最后，我们在一个全自动化的高通量学术筛选设施中实施了一项试验，说明了使用该平台设计的试验的可扩展性。这些方法上的改进简化了高度可扩展的基于神经元的表型测定的创建，旨在改善中枢神经系统疾病的药物发现。
There is a pressing need to improve approaches for drug discovery related to neuropsychiatric disorders (NSDs). Therapeutic discovery in neuropsychiatric disorders would benefit from screening assays that can measure changes in complex phenotypes linked to disease mechanisms. However, traditional assays that track complex neuronal phenotypes, such as neuronal connectivity, exhibit poor scalability and are not compatible with high-throughput screening (HTS) procedures. Therefore, we created a neuronal phenotypic assay platform that focused on improving the scalability and affordability of neuron-based assays capable of tracking disease-relevant phenotypes. First, using inexpensive laboratory-level automation, we industrialized primary neuronal culture production, which enabled the creation of scalable assays within functioning neural networks. We then developed a panel of phenotypic assays based on culturing of primary neurons from genetically modified mice expressing HTS-compatible reporters that capture disease-relevant phenotypes. We demonstrated that a library of 1,280 compounds was quickly screened against both assays using only a few litters of mice in a typical academic laboratory setting. Finally, we implemented one assay in a fully automated high-throughput academic screening facility, illustrating the scalability of assays designed using this platform. These methodological improvements simplify the creation of highly scalable neuron-based phenotypic assays designed to improve drug discovery in CNS disorders.