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

描述（由申请人提供）：尽管对有效中枢神经系统治疗的需求巨大，但进展甚微。改善中枢神经系统药物发现工作是一个紧迫的目标，因为全世界估计有15亿人患有中枢神经系统相关疾病或障碍。我们认为，开发更有效的中枢神经系统治疗方法的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。尽管在HTS筛查活动中使用神经元并不常见，但中枢神经系统疾病靶点应该在神经元中而不是细胞系中进行检测，这似乎是合乎逻辑的。我们认为，开发一种灵活的、可扩展的、与HTS兼容的基于神经元的检测开发平台将促进探针的开发，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r01驱动的研究人员与筛选中心相互作用的一个重要障碍是这些中心无法小型化和扩大基于神经元的分析。我们已经开发了一种创新的方法，将基于神经元的台式分析迁移到HTS就绪的平台，以便使对探针发现感兴趣的神经生物学家更容易使用HTS。佛罗里达斯克里普斯的神经科学部门已经与TSRI的筛选中心和Lead ID小组合作，共同努力克服这一障碍，我们已经确定了改变当前程序的方法，使基于神经元的台式分析可以小型化和自动化，从而产生能够支持数万个分子的生物和化学筛选的交钥匙分析。我们建议，神经科学领域将从一个系统中受益匪浅，该系统使基于台式神经元的检测的迁移路线小型化，然后规模化，从而导致它们在HTS探针发现活动中使用。重要的是，我们开发的系统还将为研究人员提供一套工具，用于开发基于神经元的HTS的新检测方法。我们的提案详细介绍了开发新试剂、仪器和工作流程的计划，这些计划将证明在初级神经元中开发的台式分析可以迁移到hts兼容的分析。作为原理证明，我们将把一个台式突触发生实验移植到这个支持hts的系统中。然后，我们建议将这种hts就绪的、基于神经元的检测方法用于约25,000种化合物的筛选。在初级神经元中成功实现这种规模的筛选将证明初级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作为基于神经元的HTS的通用检测开发系统。因此，该项目的结果有望为研究人员和筛选中心提供一个能够在神经元环境中支持hts级筛选的分析开发平台。因此，资助这一提案将产生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，为整个神经科学界提供服务，任何有兴趣扩大HTS活动的台式检测的人都可以使用该平台。

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.