"A Flexible and Scalable HTS Platform for CNS Probe Discovery"

描述（由申请人提供）：尽管对有效的CNS治疗剂的需求巨大，但进展甚微。改善CNS药物发现工作是一个紧迫的目标，因为全球估计有15亿人患有CNS相关疾病或病症。我们认为，更有效的CNS治疗的一个主要障碍是缺乏支持HTS水平筛选的基于神经元的探针发现平台。虽然在HTS筛选活动中使用神经元并不常见，但CNS疾病靶点应在神经元而不是细胞系中进行测定似乎是合乎逻辑的。我们认为，开发与HTS兼容的灵活且可扩展的基于神经元的检测开发平台将促进探针开发，同时也可能刺激旨在治疗各种脑部疾病的药物发现工作。阻止R 01驱动的研究者与筛查中心互动的一个重要障碍是这些中心无法扩展和扩大基于神经元的测定。我们开发了一种创新的方法，将基于神经元的台式检测迁移到HTS就绪平台，以便使HTS更容易被对探针发现感兴趣的神经生物学家使用。斯克里普斯佛罗里达的神经科学系已经与TSRI的筛选中心和铅ID组合作，以克服这一障碍，我们已经确定了改变当前程序的方法，以便基于神经元的台式测定可以小型化和自动化，以生产能够支持数万分子的生物和化学筛选的交钥匙测定。我们建议，神经科学领域将极大地受益于一个系统，使迁移路线的台式神经元为基础的测定被小型化，然后缩放，导致他们在HTS探针发现活动的使用。重要的是，我们开发的系统还将为研究人员提供一个工具集，以开发基于神经元的HTS的新检测方法。我们的提案详细说明了开发新型试剂、仪器和工作流程的计划，这些计划将证明在原代神经元中开发的台式检测可以迁移到HTS兼容的检测。作为原理的证明，我们将把台式突触发生测定迁移到这个HTS使能的系统。然后，我们建议通过对约25，000种化合物的筛选来进行这种基于神经元的HTS就绪检测。在原代神经元中成功实现这种量级的筛选将证明原代神经元可以用作药物/探针发现的通用平台，并且我们的方法可以用作基于神经元的HTS的通用测定开发系统。因此，该项目的成果有望为研究人员和筛查中心提供一个能够支持神经元环境中HTS水平筛查的检测开发平台。因此，资助这项提案将导致新的探针来调节突触发生，但也将通过提供一个检测开发平台来服务于整个神经科学界，该平台可供任何有兴趣扩大HTS活动的台式检测的人使用。

DESCRIPTION (provided by applicant): Despite an overwhelming need for effective CNS therapeutics, little progress has been made. Improving CNS drug discovery efforts is an urgent goal, as an estimated 1.5 billion people suffer from a CNS-related disease or disorder worldwide. We believe that a major roadblock toward more effective CNS therapeutics is the lack of neuron-based probe discovery platforms cable of supporting HTS-level screening. It seems logical that CNS disease targets should be assayed in neurons instead of cell-lines, though the use of neurons in HTS screening campaigns is uncommon. We argue that development of a flexible and scalable neuron-based assay development platform that is compatible with HTS would facilitate probe development, while also perhaps spurring drug discovery efforts aimed at treating a variety of brain diseases. One of the significant barriers preventing R01-driven investigators from interacting with screening centers is the inability of these centers to miniaturize and scale-up neuron-based assays. We have developed an innovative approach for migrating neuron-based benchtop assays to an HTS-ready platform in order to make HTS more accessible to neurobiologists interested in probe discovery. The Neuroscience Department at Scripps Florida has engaged the Screening Center and the Lead ID group at TSRI in collaborative efforts to overcome this barrier and we have identified ways to alter current procedures so that neuron-based benchtop assays can be miniaturized and automated to produce turnkey assays capable of supporting biological and chemical screens with tens-of-thousands of molecules. We propose that the neuroscience field would benefit tremendously from a system that enabled a migratory route for bench-top neuron-based assays to be miniaturized and then scaled, leading to their use in HTS probe discovery campaigns. Importantly, the system that we have developed will also provide investigators with a toolset to develop novel assays for neuron-based HTS. Our proposal details plans to develop novel reagents, instrumentation and workflows that will demonstrate that a bench-top assay developed in primary neurons can be migrated to an HTS-compatible assay. As a proof of principle, we will migrate a bench-top synaptogenesis assay to this HTS-enabled system. We then propose to take this HTS-ready, neuron-based assay through a screen of ~25,000 compounds. Successful achievement of a screen of this magnitude in primary neurons would demonstrate that primary neurons can be used as a common platform for drug/probe discovery and that our approach could serve as a general assay development system for neuron-based HTS. Thus, the outcome of this project is expected to provide researchers and Screening Centers with an assay development platform capable of supporting HTS-level screens in a neuronal environment. Thus, funding this proposal will result in novel probes to regulate synaptogenesis, but will also generally serve the Neuroscience community as a whole by providing an assay development platform that can be used by anyone interested in scaling up bench-top assays for HTS campaigns.