Phenotype MicroArrays for Drug Toxicity Screening

描述(由申请人提供)：在这项第二阶段的SBIR资助中，我们建议将我们的表型微阵列(PM)技术发展为药物毒性的高通量筛选(HTS)分析。该提案解决了药物发现中的一个关键需求，即需要更快、更简单、更便宜的技术来评估潜在毒理学候选药物以及作用机制。Biolog已经开发出一种原型技术，可以用于药物毒性的体外筛选。它的设计是为了与选定的人类细胞系合作，以代表通常表现出毒性的不同器官和组织。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培养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。这种基于细胞的毒理学测试方法的一个独特方面是，我们可以在近2000种不同的细胞生理状态下看到药物对细胞的影响。测试细胞或动物的替代方法只测试健康和增殖状态下的毒理学。另一个独特的方面是，该技术还可以测试药物组合，并可以提供非常有用的协同和拮抗信息。在这个项目的第二阶段，我们建议将PM测试的范围从目前的600个扩大到大约2000个，将技术小型化和自动化，然后通过测试2000种已知类别和毒性的化学物质来构建一个初步的化学指纹数据库。除了在毒理学方面的应用外，这项技术还将用于研究细胞生物学、代谢性疾病和基因功能的基础研究，以及用于细胞系质量控制测试和生物过程开发的应用细胞生物学。

DESCRIPTION (provided by applicant): In this Phase II SBIR grant, we propose to develop our Phenotype MicroArray (PM) technology into a high throughput screening (HTS) assay for drug toxicity. The proposal addresses a key need in drug discovery for faster, simpler, and less expensive technologies that can assess drug candidates for potential toxicology as well as mechanism of action. Biolog has developed a prototype technology that can be employed for in vitro screening of drug toxicity. It is designed to work with human cell lines selected to represent the different organs and tissues where toxicity is typically exhibited. The 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 cultured 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. A unique aspect of this cell-based approach to toxicological testing is that we can see the effect of the drug on cells under nearly 2000 diverse physiological states of the cell. Alternative methods of testing cells or animals only test for toxicology in a healthy and proliferative state. Another unique aspect is that the technology also tests drug combinations and can provide highly useful synergy and antagonism information. In Phase II of this project we propose to expand the set of PM tests that can be performed from the current set of 600 to approximately 2000, miniaturize and automate the technology, and then construct an initial database of chemical fingerprints by testing 2000 chemicals of known classes and toxicities. In addition to applications in toxicology, this technology will be useful in basic research to study cell biology, metabolic diseases, and gene function, and in applied cell biology for QC testing of cell lines and bioprocess development.