Fishing for Teratogens: A Consortium Effort for a Harmonized Zebrafish Developmental Toxicology Assay
Fishing for Teratogens: A Consortium Effort for a Harmonized Zebrafish Developmental Toxicology Assay
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DOI：
10.1093/toxsci/kfu017
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发表时间：
2014-05-01
影响因子：
3.8
通讯作者：
Augustine-Rauch, Karen A.
中科院分区：
文献类型：
--
作者：
Ball, Jonathan S.;Stedman, Donald B.;Augustine-Rauch, Karen A.
生物制药公司联盟此前开发了一种优化的斑马鱼发育毒性测定 (ZEDTA)，其中将绒毛膜胚胎在受精后 5 至 6 小时内暴露于非专有测试化合物，并在受精后 5 天评估形态完整性。使用最初的 20 种测试化合物，对哺乳动物体内结果的致畸性达到了 88% 的总体预测值 [Gustafson, A. L.、Stedman, D. B.、Ball, J.、Hillegass, J. M.、Flood, A.、Zhang, C. X.、Panzica-Kelly, J.、Cao, J.、Coburn, A.、Enright, B. P. 等人。 （2012）。协调斑马鱼发育毒理学测定的实验室间评估-第一阶段进展报告。再现。毒性。 33、155-164]。在该项目的第二阶段，来自四个联盟成员的 38 种专有药物化合物在两个实验室中使用优化方法进行了评估，使用池塘来源或培养品系的野生型斑马鱼胚胎，浓度高达 100 μM。使用液相色谱-串联质谱法评估了胚胎对所有化合物的吸收。 38 种化合物中的 28 种经证实胚胎摄取率 > 5%，并且 ZEDTA 在两个实验室分别实现了这些化合物 82% 和 65% 的总体预测值。当以高达 1000 μM 的对数浓度重新测试低摄取化合物（千分之一货币符号 5%）时，所有 38 种化合物的总体预测率分别为 79% 和 62%，第一个实验室实现了 74% 的灵敏度（致畸剂检测）和 82% 的特异性（非致畸剂检测），第二个实验室实现了 63% 的灵敏度（致畸剂检测）和 62% 的特异性（非致畸检测）。随后的数据分析表明，技术差异而不是应变差异是造成实验室间预测差异的主要原因。基于这些结果，目前 4/5 的联盟公司正在使用 ZEDTA 统一方法进行先导化合物开发优化阶段的化合物评估。
A consortium of biopharmaceutical companies previously developed an optimized Zebrafish developmental toxicity assay (ZEDTA) where chorionated embryos were exposed to non-proprietary test compounds from 5 to 6 h post fertilization and assessed for morphological integrity at 5 days post fertilization. With the original 20 test compounds, this achieved an overall predictive value for teratogenicity of 88% of mammalian in vivo outcome [Gustafson, A. L., Stedman, D. B., Ball, J., Hillegass, J. M., Flood, A., Zhang, C. X., Panzica-Kelly, J., Cao, J., Coburn, A., Enright, B. P., et al. (2012). Interlaboratory assessment of a harmonized Zebrafish developmental toxicology assay-Progress report on phase I. Reprod. Toxicol. 33, 155-164]. In the second phase of this project, 38 proprietary pharmaceutical compounds from four consortium members were evaluated in two laboratories using the optimized method using either pond-derived or cultivated-strain wild-type Zebrafish embryos at concentrations up to 100 mu M. Embryo uptake of all compounds was assessed using liquid chromatography-tandem mass spectrometry. Twenty eight of 38 compounds had a confirmed embryo uptake of > 5%, and with these compounds the ZEDTA achieved an overall predictive value of 82% and 65% at the two respective laboratories. When low-uptake compounds (a parts per thousand currency sign 5%) were retested with logarithmic concentrations up to 1000 mu M, the overall predictivity across all 38 compounds was 79% and 62% respectively, with the first laboratory achieving 74% sensitivity (teratogen detection) and 82% specificity (non-teratogen detection) and the second laboratory achieving 63% sensitivity (teratogen detection) and 62% specificity (non-teratogen detection). Subsequent data analyses showed that technical differences rather than strain differences were the primary contributor to interlaboratory differences in predictivity. Based on these results, the ZEDTA harmonized methodology is currently being used for compound assessment at lead optimization stage of development by 4/5 of the consortium companies.