Comprehensive Analyses and Prioritization of Tox21 10K Chemicals Affecting Mitochondrial Function by in-Depth Mechanistic Studies.
Comprehensive Analyses and Prioritization of Tox21 10K Chemicals Affecting Mitochondrial Function by in-Depth Mechanistic Studies.
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DOI：
10.1289/ehp2589
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发表时间：
2018-07
影响因子：
10.4
通讯作者：
Simeonov A
中科院分区：
文献类型：
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作者：
Xia M;Huang R;Shi Q;Boyd WA;Zhao J;Sun N;Rice JR;Dunlap PE;Hackstadt AJ;Bridge MF;Smith MV;Dai S;Zheng W;Chu PH;Gerhold D;Witt KL;DeVito M;Freedman JH;Austin CP;Houck KA;Thomas RS;Paules RS;Tice RR;Simeonov A
毒性测试的一个主要挑战是商业中的大量化学品缺乏毒理学评估。作为回应，Tox21 计划正在将毒性测试的重点从动物研究重新转向使用目标/途径特异性、机制驱动的检测的更便宜、更高通量的体外方法。我们的目标是使用深入的机制研究方法来优先考虑和表征影响线粒体功能的化学物质。我们使用分层测试方法，优先对 622 种化合物进行更广泛的测试，这些化合物是通过对 8,300 种独特小分子（包括药物和工业化学品）进行初级定量高通量筛选而鉴定出的，这些化合物是潜在的线粒体毒物，因为它们能够显着降低线粒体膜电位 (MMP)。根据 HepG2 细胞和大鼠肝细胞的二级 MMP 测定结果，选择了 34 种化合物进行三级测定，包括线虫线虫中活性氧 (ROS) 的形成、p53 和核红细胞 2 相关因子 2/抗氧化反应元件 (Nrf2/ARE) 的上调、线粒体耗氧量、细胞 Parkin 易位以及幼虫发育和 ATP 状态线虫。检测到一组已知的线粒体复合物抑制剂（例如鱼藤酮）和解偶联剂（例如氯芬那吡），以及潜在的新型复合物抑制剂和解偶联剂。从这项研究中，我们确定了四种尚未充分表征的潜在线粒体毒物（拉沙洛菌素、啶氧菌酯、频氰醇和三氯卡班），值得进一步进行体内表征。用于识别和机械表征线粒体毒物的分层方法可能会减少毒理学测试中动物的使用。 https://doi.org/10.1289/EHP2589
A central challenge in toxicity testing is the large number of chemicals in commerce that lack toxicological assessment. In response, the Tox21 program is re-focusing toxicity testing from animal studies to less expensive and higher throughput in vitro methods using target/pathway-specific, mechanism-driven assays. Our objective was to use an in-depth mechanistic study approach to prioritize and characterize the chemicals affecting mitochondrial function. We used a tiered testing approach to prioritize for more extensive testing 622 compounds identified from a primary, quantitative high-throughput screen of 8,300 unique small molecules, including drugs and industrial chemicals, as potential mitochondrial toxicants by their ability to significantly decrease the mitochondrial membrane potential (MMP). Based on results from secondary MMP assays in HepG2 cells and rat hepatocytes, 34 compounds were selected for testing in tertiary assays that included formation of reactive oxygen species (ROS), upregulation of p53 and nuclear erythroid 2-related factor 2/antioxidant response element (Nrf2/ARE), mitochondrial oxygen consumption, cellular Parkin translocation, and larval development and ATP status in the nematode Caenorhabditis elegans. A group of known mitochondrial complex inhibitors (e.g., rotenone) and uncouplers (e.g., chlorfenapyr), as well as potential novel complex inhibitors and uncouplers, were detected. From this study, we identified four not well-characterized potential mitochondrial toxicants (lasalocid, picoxystrobin, pinacyanol, and triclocarban) that merit additional in vivo characterization. The tier-based approach for identifying and mechanistically characterizing mitochondrial toxicants can potentially reduce animal use in toxicological testing. https://doi.org/10.1289/EHP2589