Phenotype MicroArrays for Drug Toxicity Screening

描述(由申请人提供)： Biolog已经开发出表型微阵列(PM)，可以用作药物毒性的体外筛选。这项技术可以以简单易用的HTS格式测量细胞的数千个生理参数。这项技术的核心是一种化学和图像分析系统，它可以检测在不同生理条件下生长的细胞呼吸活动的变化。细胞代谢活动的变化是通过Biolog的自动OmniLog仪器检测到的比色反应来测量的。PMS允许为所检查的每一种药物和细胞系生成信息丰富的比色模式。当一种药物在PM中测试时，比色模式发生变化，使用模式识别技术，我们可以将PM中的颜色变化与药物的作用模式和副作用联系起来。这个图案是该药物作用的指纹。当我们测试大量具有已知药理作用的药物时，每种药物的个人指纹将具有共性，这将使我们能够确定该类药理制剂的参考标准。这些特征的数据库可用于在HTS程序中对新化合物进行分类。重要的是，通过测试具有众所周知的体内毒性的药物，指纹将拥有有关被测试药物的毒理学信息，并可用于预测一种药物在以后的开发中是否具有毒性。在第一阶段SBIR赠款中，我们建议将我们的PM技术发展为HTS药物毒性测试。对药物毒性最敏感的两种组织是肝脏和血液，它们可以很好地作为药物毒性潜力的晴雨表。在这笔赠款中，我们将为人类肝脏和造血细胞株优化新的PM原型和测试方案。然后，我们将测试一系列经过充分研究的药物，这些药物在结构和机制上是不同的，但在体内具有已知的毒性和副作用。这些药物产生的代谢活动的概况将被整理，以产生一个数据库的原型，该数据库可用于筛选小分子的毒性潜力。在未来的研究中(第二阶段SBIR)，我们将扩大参比药物测试的技术和范围，以建立一个强大的预测性数据库，包括代表更多组织和器官的细胞系。

DESCRIPTION (provided by applicant): Biolog has developed Phenotype MicroArrays (PMs) that can be employed as an in vitro screen for drug toxicity. This technology can measure thousands of physiological parameters of a cell in a simple-to-use HTS format. The core of the technology is a chemistry and image analysis system that detects changes in respiratory activity of cells grown under different physiological conditions. The changes in metabolic activity of the cell are measured as a colorimetric response detected with Biolog's automated OmniLog instrument. PMs allow for an information-rich colorimetric pattern to be generated for each drug and cell line examined. When a drug is tested in the PM, the colorimetric pattern changes and using pattern recognition technology, we can relate the color changes in the PM to the mode of action and side effects of the drug. The pattern is a fingerprint of that drug's action. When we test a large number of drugs with known pharmacological action, the individual fingerprints of each drug will have commonalities that will allow us to identify a reference standard for that class of pharmacological agents. A database of these signatures can be used to categorize novel compounds in a HTS program. Importantly, by testing drugs with well known in vivo toxicity, the fingerprints will have information about the toxicology of the drug tested and can be used to predict whether a drug "hit" will be toxic later on in development. In this Phase I SBIR grant, we propose to develop our PM technology into a HTS assay for drug toxicity. Two tissues which are most susceptible to drug toxicity are the liver and blood and these can serve as a good barometer for drug toxicity potential. In this grant we will optimize new prototype PMs and testing protocols for human liver and hematopoietic cell lines. We will then test a series of well-studied drugs that are structurally and mechanistically distinct but which have known toxicity and side effects in vivo. The profiles of metabolic activity generated by these drugs will be collated to generate a prototype of a database that can be used to screen small molecules for toxicity potential. In future studies (Phase II SBIR) we will expand the technology and scope of reference drugs tested to build a strong predictive database including cell lines representing more tissues and organs.