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

描述(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。需要新的中枢神经系统药物，但神经系统的复杂性在很大程度上使还原和体外方法难以发现中枢神经系统药物。因此，大多数现有的中枢神经系统药物是通过对活体动物行为影响的偶然观察而发现的，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的神经活性药物的手段。我们已经开发了一种全自动系统，能够以高通量、96孔格式跟踪和量化斑马鱼的行为。来自特定功能类别的精神药物在本实验中产生不同的行为特征，表明斑马鱼的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基因扰动的训练集来测试和验证该分析。然后，5万个小分子的筛选将识别出新的神经活性化合物，这些化合物产生的行为特征与训练集中已知的精神药物的行为特征相匹配。我们开发的斑马鱼测试是第一个能够评估脊椎动物小分子行为影响的高通量筛查。由于斑马鱼的行为整合了几个主要神经递质系统的输入，该测试可以用来识别通过不同机制作用于中枢神经系统的化合物。我们正在使用的800种精神药物的广泛训练集使该分析得以仔细验证，并提供了一种强大的手段，为发现的新化合物分配潜在的作用机制。该项目的成功完成将创建一个强大而灵活的系统来发现神经活性化合物。它还将直接导致发现新的先导化合物，这些化合物可能会进一步开发用于治疗神经系统疾病。与公共健康相关：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，以及由此产生的需要在困难的动物行为测试中测试候选药物。我们已经开发了一种简单的、自动化的动物行为测试，并将使用该测试来快速筛选数千种候选化合物，并识别那些有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.