In Vivo Discovery of Psychotropic Drugs by High-Throughput Behavioral Phenotyping

通过高通量行为表型在体内发现精神药物的挑战尽管它们无处不在和影响，中枢神经系统（CNS）疾病仍然是治疗最差的医疗条件之一。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得中枢神经系统药物的发现难以还原和体外方法。因此，大多数现有的中枢神经系统药物都是通过对活体动物行为效应的偶然观察发现的，而不是通过合理设计或体外筛选发现的。发现新的中枢神经系统药物受到体外复杂脑功能建模的困难以及用现有的哺乳动物行为分析在体内筛选新药的不可行性的限制。我们的方法，我们建议使用高通量行为分析作为一种手段筛选新的神经活性药物。我们正在开发全自动系统，能够以高通量、96孔格式跟踪和量化斑马鱼的行为。使用其中一种分析方法，我们测试了700种来自几种功能类别的精神药物，并确定了特定功能类别与它们诱导的斑马鱼行为特征之间的强烈相关性。我们现在建议扩大自动化行为分析的范围，并对大量小分子文库进行筛选，以识别具有体内神经活动的新化合物。我们正在开发的自动化斑马鱼检测面板的潜在影响是第一个能够评估脊椎动物小分子行为影响的高通量筛选。因为斑马鱼的行为整合了来自几个主要神经递质系统的输入，所以这些检测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。这个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。它还将直接导致通过不同的作用机制改变中枢神经系统功能的新化合物的发现。这些化合物将成为研究神经系统的有力工具，在某些情况下可能进一步开发用于治疗神经系统疾病。精神分裂症和阿尔茨海默病等神经系统疾病普遍存在，而且往往具有毁灭性，但由于传统的药物发现方法无法应对大脑的复杂性，这些疾病的治疗仍然很差。该项目提出了一种大胆的神经系统药物发现新方法，该方法基于机器人对数千种潜在新药的测试，以了解它们改变微观斑马鱼大脑功能的能力。

In vivo discovery of psychotropic drugs by high-throughput behavioral phenotyping the challenge despite their ubiquity and impact, diseases of the central nervous system (CNS) remain among the most poorly treated medical conditions. 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 rational design or in vitro screening. Discovering new CNS drugs is limited by the difficulty of modeling complex brain function in vitro and the impracticality of screening for new drugs in vivo with existing mammalian behavioral assays. Our approach we propose to use high-throughput behavioral assays as a means of screening for novel neuroactive drugs. We are developing fully-automated systems capable of tracking and quantifying zebrafish behaviors in high- throughput, 96-well format. Using one of these assays, we have tested 700 psychotropic drugs from several functional classes and identified strong correlations between specific functional classes and the zebrafish behavioral profiles they induce. We now propose to expand the panel of automated behavioral assays and conduct screens of vast small molecule libraries to identify novel compounds with in vivo neurological activity. The potential impact the automated panel of zebrafish assays we are developing is the first high-throughput screen capable of assessing behavioral effects of small molecules in a vertebrate. Because the zebrafish behaviors integrate inputs from several major neurotransmitter systems, the assays can be used to identify compounds that act on the CNS through diverse mechanisms. 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 compounds that alter CNS function through diverse mechanisms of action. These compounds will be powerful tools for studying the nervous system and in some cases may be developed further for treating nervous system disorders. Nervous system disorders like schizophrenia and Alzheimer's disease are widespread and frequently devastating, but they remain poorly treated because conventional drug discovery methods are poorly equipped to deal with the complexity of the brain. This project proposes a bold new approach to nervous system drug discovery based on robotic testing of thousands of potential new drugs for their ability to alter brain function in microscopic zebrafish.