High-throughput behavioral screening for in vivo discovery of neuroactive drugs

描述（由申请人提供）：中枢神经系统（CNS）疾病包括许多最具破坏性和负担的所有人类疾病。尽管神经系统疾病无处不在，影响巨大，但它们仍然是治疗最差的疾病之一。需要新的CNS药物，但神经系统的复杂性在很大程度上使CNS药物发现难以还原和体外方法。由于这个原因，大多数现有的中枢神经系统药物是通过偶然观察活体动物的行为效应而发现的，而不是通过体外筛选。我们建议使用高通量行为分析作为筛选新型神经活性药物的一种手段。我们已经开发了一个全自动的系统，能够跟踪和量化斑马鱼的行为在高通量，96孔格式。来自特定功能类别的精神药物在该测定中产生不同的行为特征，表明斑马鱼行为特征与小分子作用机制之间存在很强的相关性。我们的目标是使用800种已知神经活性药物和特定遗传扰动的训练集来测试和验证该测定。然后，对5万个小分子进行筛选，将识别出新的神经活性化合物，这些化合物在训练集中产生与已知精神药物相匹配的行为特征。我们开发的斑马鱼检测是第一个能够评估脊椎动物中小分子行为影响的高通量筛选。由于斑马鱼的行为整合了来自几个主要神经递质系统的输入，因此该测定可用于鉴定通过不同机制作用于CNS的化合物。我们正在使用的800种精神药物的广泛训练集能够仔细验证该测定，并为发现的新化合物分配潜在的作用机制提供了强有力的手段。该项目的成功完成将为发现神经活性化合物创造一个强大而灵活的系统。它还将直接导致发现新的先导化合物，可能进一步开发用于治疗神经系统疾病。公共卫生相关性：发现神经系统疾病的新疗法一直很困难，因为大脑的复杂性和由此产生的需要在困难的动物行为测试中测试候选药物。我们开发了一种简单的动物行为自动测试，并将使用该测试快速筛选数千种候选化合物，并确定那些有希望治疗神经系统疾病的化合物。

DESCRIPTION (provided by applicant): Diseases of the central nervous system (CNS) include many of the most devastating and burdensome of all human illnesses. Despite their ubiquity and impact, diseases of the nervous system remain among the most poorly treated. New CNS drugs are needed, but the complexity of the nervous system has largely made CNS drug discovery refractory to reductionist and in vitro approaches. For this reason, most existing CNS drugs were discovered by serendipitous observation of behavioral effects in living animals, not by in vitro screening. We propose to use high-throughput behavioral assays as a means of screening for novel neuroactive drugs. We have developed a fully-automated system capable of tracking and quantifying zebrafish behaviors in high- throughput, 96-well format. Psychotropic drugs from specific functional classes produce distinct behavioral profiles in this assay, suggesting a strong correlation between zebrafish behavioral profiles and small molecule mechanism of action. We aim to use a training set of 800 known neuroactive drugs and specific genetic perturbations to test and validate the assay. A screen of 50,000 small molecules will then identify novel neuroactive compounds that create behavioral profiles matching those of known psychotropic drugs in the training set. The zebrafish assay we have developed is the first high-throughput screen capable of assessing behavioral effects of small molecules in a vertebrate. Because the zebrafish behavior integrates inputs from several major neurotransmitter systems, the assay can be used to identify compounds that act on the CNS through diverse mechanisms. The extensive training set of 800 psychotropic drugs we are using enables careful validation of the assay and provides a powerful means of assigning potential mechanisms of action for novel compounds discovered. Successful completion of this project will create a robust and flexible system for discovering neuroactive compounds. It will also lead directly to discovery of novel lead compounds that may be developed further for treating nervous system disorders. PUBLIC HEALTH RELEVANCE: Discovering new treatments for diseases of the nervous system has been difficult because of the brain's complexity and the resulting need to test drug candidates in difficult animal behavioral tests. We have developed a simple, automated test of animal behaviors and will use the test to screen rapidly through thousands of candidate compounds and identify those with promise for treating nervous system disorders.