Characterization of diversity in toxicity mechanism using in vitro cytotoxicity assays in quantitative high throughput screening
Characterization of diversity in toxicity mechanism using in vitro cytotoxicity assays in quantitative high throughput screening
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DOI：
10.1021/tx700365e
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发表时间：
2008-03-01
影响因子：
4.1
通讯作者：
Austin, Christopher P.
中科院分区：
文献类型：
--
作者：
Huang, Ruili;Southall, Noel;Austin, Christopher P.
评估环境化合物的潜在健康风险是一项昂贵的工作，这推动了传统体内毒理学测试的新替代品的开发。一种方法是分阶段进行评估，从成本更低、吞吐量更高的体外测试开始，然后进行更明确的试验。体外试验可以用来产生关于化合物作用机制的假设，然后可以用来设计适当的体内实验。在这里，我们开始解决如何设计这样一个基于细胞的体外分析电池的问题，通过结合两种不同类型的检测数据，细胞存活率和caspase激活，目的是阐明作用机制。由于caspase激活是细胞凋亡过程中的一个瞬时事件，因此不可能设计一个单一的终点检测方法来识别化合物诱导的caspase激活的所有实例。然而，结合细胞存活率数据，可以从这些检测中获得有关化合物作用机制的有用信息。非监督聚类结合Dunn的聚类有效性指数是一种稳健的方法，可以识别作用机制，而不需要任何关于毒性机制的先验知识。通过将聚类结果与复合机构的文献注释进行比较，评估了该聚类方法的性能。
Assessing the potential health risks of environmental chemical compounds is an expensive undertaking that has motivated the development of new alternatives to traditional in vivo toxicological testing. One approach is to stage the evaluation, beginning with less expensive and higher throughput in vitro testing before progressing to more definitive trials. In vitro testing can be used to generate a hypothesis about a compound's mechanism of action, which can then be used to design an appropriate in vivo experiment. Here we begin to address the question of how to design such a battery of in vitro cell-based assays by combining data from two different types of assays, cell viability and caspase activation, with the aim of elucidating the mechanism of action. Because caspase activation is a transient event during apoptosis, it is not possible to design a single end-point assay protocol that would identify all instances of compound-induced caspase activation. Nevertheless, useful information about compound mechanism of action can be obtained from these assays in combination with cell viability data. Unsupervised clustering in combination with Dunn's cluster validity index is a robust method for identifying mechanisms of action without requiring any a priori knowledge about mechanisms of toxicity. The performance of this clustering method is evaluated by comparing the clustering results against literature annotations of compound mechanisms.