Real-time cell toxicity profiling of Tox21 10K compounds reveals cytotoxicity dependent toxicity pathway linkage.
Real-time cell toxicity profiling of Tox21 10K compounds reveals cytotoxicity dependent toxicity pathway linkage.
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DOI：
10.1371/journal.pone.0177902
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Auerbach SS
中科院分区：
文献类型：
--
作者：
Hsieh JH;Huang R;Lin JA;Sedykh A;Zhao J;Tice RR;Paules RS;Xia M;Auerbach SS
细胞毒性是评价化学毒性的常用体外终点。为了支持美国Tox 21筛选计划，使用两种多重实时测定技术，以浓度依赖性方式在两种细胞系（HEK 293、HepG 2）中以0、8、16、24、32和40小时暴露时研究~ 10 K化学品的细胞毒性。一种技术测量细胞的代谢活性（即，细胞活力，GLO）而另一种评价细胞膜完整性（即，细胞死亡，flor）。使用glo技术，在HEK 293细胞中观察到更多的活性物质和更大的时间变化，而flor技术的结果在两种细胞类型中更相似。根据化学品的细胞毒性动力学特征将其分组，并评价这些类别与Tox 21核受体和应激反应途径试验中活性的相关性。一些途径，如H2 AX的激活，与快速响应的细胞毒性类别相关，而其他途径，如TP 53的激活，与缓慢响应的细胞毒性类别相关。通过基于其与不同细胞毒性动力学标签的关联程度对途径进行聚类，我们确定了活性化学品呈现相似细胞毒性动力学的途径簇。这种联系可能是由于途径之间共享的潜在生物学过程，例如，H2 AX和热休克因子的激活。其他涉及核受体活性的可能是由于共享的化学结构，而不是途径水平的相互作用。基于雄激素受体拮抗作用和Nrf 2活性之间的联系，我们推测雄激素受体拮抗剂的一个亚类通过与Nrf 2活化相关的氧化应激引起细胞毒性。总之，实时细胞毒性筛选提供了与其细胞毒性机制相关的信息丰富的化学细胞毒性动力学数据，并且通过我们的分析，可以制定关于受试化学品细胞毒性特性的基于机制的假设。
Cytotoxicity is a commonly used in vitro endpoint for evaluating chemical toxicity. In support of the U.S. Tox21 screening program, the cytotoxicity of ~10K chemicals was interrogated at 0, 8, 16, 24, 32, & 40 hours of exposure in a concentration dependent fashion in two cell lines (HEK293, HepG2) using two multiplexed, real-time assay technologies. One technology measures the metabolic activity of cells (i.e., cell viability, glo) while the other evaluates cell membrane integrity (i.e., cell death, flor). Using glo technology, more actives and greater temporal variations were seen in HEK293 cells, while results for the flor technology were more similar across the two cell types. Chemicals were grouped into classes based on their cytotoxicity kinetics profiles and these classes were evaluated for their associations with activity in the Tox21 nuclear receptor and stress response pathway assays. Some pathways, such as the activation of H2AX, were associated with the fast-responding cytotoxicity classes, while others, such as activation of TP53, were associated with the slow-responding cytotoxicity classes. By clustering pathways based on their degree of association to the different cytotoxicity kinetics labels, we identified clusters of pathways where active chemicals presented similar kinetics of cytotoxicity. Such linkages could be due to shared underlying biological processes between pathways, for example, activation of H2AX and heat shock factor. Others involving nuclear receptor activity are likely due to shared chemical structures rather than pathway level interactions. Based on the linkage between androgen receptor antagonism and Nrf2 activity, we surmise that a subclass of androgen receptor antagonists cause cytotoxicity via oxidative stress that is associated with Nrf2 activation. In summary, the real-time cytotoxicity screen provides informative chemical cytotoxicity kinetics data related to their cytotoxicity mechanisms, and with our analysis, it is possible to formulate mechanism-based hypotheses on the cytotoxic properties of the tested chemicals.