Systematic study of mitochondrial toxicity of environmental chemicals using quantitative high throughput screening.
Systematic study of mitochondrial toxicity of environmental chemicals using quantitative high throughput screening.
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DOI：
10.1021/tx4001754
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发表时间：
2013-09-16
影响因子：
4.1
通讯作者：
Xia M
中科院分区：
文献类型：
--
作者：
Attene-Ramos MS;Huang R;Sakamuru S;Witt KL;Beeson GC;Shou L;Schnellmann RG;Beeson CC;Tice RR;Austin CP;Xia M
Tox21计划的目标是将毒性测试从传统的体内模型过渡到体外试验，以评估化学品如何影响细胞反应和毒性途径。美国国立卫生研究院化学基因组学中心（NCGC）对Tox21计划的关键贡献是实施定量高通量筛选（qHTS）方法，使用基于细胞和生物化学的测定来生成数千种环境化合物的毒理学特征。在这里，我们通过在qHTS平台中筛选国家毒理学计划（NTP）提供的1，408种化合物的库，评估了化合物对HepG2细胞线粒体膜电位的影响。对14种浓度的化合物进行了筛选，结果显示，处理1小时或5小时后，分别有91种和88种化合物破坏线粒体膜电位。在两个时间点活性的76种化合物通过结构相似性进行聚类，产生11个簇和23个单体。38种化合物涵盖了大部分的活性化学空间进行了更广泛的评价。38种化合物中的36种使用荧光板读数器确认破坏线粒体膜电位，35种使用高含量成像方法确认。在38种化合物中，4种和6种分别在1小时或5小时诱导LDH释放，这是细胞毒性的量度。通过测量耗氧速率的变化进一步评估化合物的作用机制（MOA），这使得能够鉴定20种化合物作为解偶联剂。这种全面的方法允许评估数千种环境化学品的线粒体毒性和识别可能的MOA。
A goal of the Tox21 program is to transit toxicity testing from traditional in vivo models to in vitro assays that assess how chemicals affect cellular responses and toxicity pathways. A critical contribution of the NIH Chemical Genomics center (NCGC) to the Tox21 program is the implementation of a quantitative high throughput screening (qHTS) approach, using cell- and biochemical-based assays to generate toxicological profiles for thousands of environmental compounds. Here, we evaluated the effect of chemical compounds on mitochondrial membrane potential in HepG2 cells by screening a library of 1,408 compounds provided by the National Toxicology Program (NTP) in a qHTS platform. Compounds were screened over 14 concentrations, and results showed that 91 and 88 compounds disrupted mitochondrial membrane potential after treatment for one or five h, respectively. Seventy-six compounds active at both time points were clustered by structural similarity, producing 11 clusters and 23 singletons. Thirty-eight compounds covering most of the active chemical space were more extensively evaluated. Thirty-six of the 38 compounds were confirmed to disrupt mitochondrial membrane potential using a fluorescence plate reader and 35 were confirmed using a high content imaging approach. Among the 38 compounds, 4 and 6 induced LDH release, a measure of cytotoxicity, at 1 or 5 h, respectively. Compounds were further assessed for mechanism of action (MOA) by measuring changes in oxygen consumption rate, which enabled identification of 20 compounds as uncouplers. This comprehensive approach allows for evaluation of thousands of environmental chemicals for mitochondrial toxicity and identification of possible MOAs.
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DOI：
10.1080/10937404.2010.483176
发表时间：
2010-02
期刊：
Journal of toxicology and environmental health. Part B, Critical reviews
影响因子：
--
作者：
Krewski D;Acosta D Jr;Andersen M;Anderson H;Bailar JC 3rd;Boekelheide K;Brent R;Charnley G;Cheung VG;Green S Jr;Kelsey KT;Kerkvliet NI;Li AA;McCray L;Meyer O;Patterson RD;Pennie W;Scala RA;Solomon GM;Stephens M;Yager J;Zeise L
通讯作者：
Zeise L
DOI：
10.1073/pnas.59.2.484
发表时间：
1968-01-01
影响因子：
11.1
作者：
HOPFER, U;LEHNINGER, AL;THOMPSON, TE
通讯作者：
THOMPSON, TE
DOI：
10.1111/j.1365-2672.1974.tb00422.x
发表时间：
1974-01-01
期刊：
JOURNAL OF APPLIED BACTERIOLOGY
影响因子：
--
作者：
BLOOMFIELD, SF
通讯作者：
BLOOMFIELD, SF
DOI：
10.1111/j.1432-1033.1970.tb00828.x
发表时间：
1970-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
LECHNER, V;SIESS, M;HOFFMANN, PC
通讯作者：
HOFFMANN, PC
影响因子：
3
作者：
Kowaltowski, AJ;Turin, J;Vercesi, AE
通讯作者：
Vercesi, AE