High-content, high-throughput screening for the identification of cytotoxic compounds based on cell morphology and cell proliferation markers.
High-content, high-throughput screening for the identification of cytotoxic compounds based on cell morphology and cell proliferation markers.
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DOI：
10.1371/journal.pone.0088338
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Tomlinson DC
中科院分区：
文献类型：
--
作者：
Martin HL;Adams M;Higgins J;Bond J;Morrison EE;Bell SM;Warriner S;Nelson A;Tomlinson DC
毒性是药物发现和开发失败的主要原因，虽然进行了强有力的毒理学测试，但如果在初级化合物筛选过程中鉴定出具有细胞毒性特征的化合物，则可以提高效率。高内涵成像在初级筛选中的使用越来越广泛，通过利用表型方法，应该可以将细胞毒性反筛选纳入初级筛选中。在这里，我们提出了一种新颖的表型测定，可用作反筛选来识别具有不利细胞效应的化合物。该测定是使用 U2OS 细胞、PerkinElmer Operetta 高内涵/高通量成像系统和 Columbus 图像分析软件开发的。在哥伦布，设计了分别使用 DAPI、TOTO-3 和磷酸组蛋白 H3 染色来识别核形态、细胞形状和增殖变化的算法。该算法是在用阿霉素、紫杉醇和诺考达唑处理的细胞上开发和测试的。然后，该测定被用来筛选一个新颖的化学库，该库富含 300 多种化合物的类似天然产物的分子，其中 13.6% 被鉴定为具有不利的细胞效应。该测定提供了一种相对便宜且快速的方法，用于在筛选测定期间鉴定具有不良细胞效应的化合物，可能减少后续体外和体内测定中由于毒性而导致的化合物排斥。
Toxicity is a major cause of failure in drug discovery and development, and whilst robust toxicological testing occurs, efficiency could be improved if compounds with cytotoxic characteristics were identified during primary compound screening. The use of high-content imaging in primary screening is becoming more widespread, and by utilising phenotypic approaches it should be possible to incorporate cytotoxicity counter-screens into primary screens. Here we present a novel phenotypic assay that can be used as a counter-screen to identify compounds with adverse cellular effects. This assay has been developed using U2OS cells, the PerkinElmer Operetta high-content/high-throughput imaging system and Columbus image analysis software. In Columbus, algorithms were devised to identify changes in nuclear morphology, cell shape and proliferation using DAPI, TOTO-3 and phosphohistone H3 staining, respectively. The algorithms were developed and tested on cells treated with doxorubicin, taxol and nocodazole. The assay was then used to screen a novel, chemical library, rich in natural product-like molecules of over 300 compounds, 13.6% of which were identified as having adverse cellular effects. This assay provides a relatively cheap and rapid approach for identifying compounds with adverse cellular effects during screening assays, potentially reducing compound rejection due to toxicity in subsequent in vitro and in vivo assays.
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影响因子：
--
作者：
Ghosh, RN;DeBiasio, R;Oakley, RH
通讯作者：
Oakley, RH
DOI：
10.1016/j.vascn.2013.08.001
发表时间：
2013-11-01
影响因子：
1.9
作者：
Persson, Mikael;Loye, Anni F.;Hornberg, Jorrit J.
通讯作者：
Hornberg, Jorrit J.
影响因子：
3.8
作者：
Lin, Zhiwu;Will, Yvonne
通讯作者：
Will, Yvonne
影响因子：
4.3
作者：
O'Leary-Steele, Catherine;Pedersen, Palle J.;Nelson, Adam
通讯作者：
Nelson, Adam
影响因子：
3.2
作者：
Suzuki, T;Fujikura, K;Takata, K
通讯作者：
Takata, K