Validation, Optimization, and Application of the Zebrafish Developmental Toxicity Assay for Pharmaceuticals Under the ICH S5(R3) Guideline.
Validation, Optimization, and Application of the Zebrafish Developmental Toxicity Assay for Pharmaceuticals Under the ICH S5(R3) Guideline.
复制标题
DOI：
10.3389/fcell.2021.721130
复制
发表时间：
2021
影响因子：
5.5
通讯作者：
Zhang LJ
中科院分区：
文献类型：
--
作者：
Song YS;Dai MZ;Zhu CX;Huang YF;Liu J;Zhang CD;Xie F;Peng Y;Zhang Y;Li CQ;Zhang LJ
斑马鱼作为发育毒性测试的替代动物模型已被广泛研究，但其测定方案尚未统一。这项研究验证并优化了美国和欧洲多个实验室间研究先前报告的斑马鱼发育毒性测定。在本研究中，使用该经典方案，在31种ICH阳性化合物中，23种化合物（74.2%）对斑马鱼具有致畸性，5种具有假阴性结果，根据方案标准，3种既不是致畸性也不是非致畸性；在 14 种 ICH 阴性化合物中，12 种化合物 (85.7%) 对斑马鱼无致畸作用，其中两种化合物出现假阳性结果。我们在用 2 dpf 测试化合物处理的斑马鱼和原来的 5 dpf 的基础上添加了一个额外的 TI 值，提出了一个新的类别作为未分类的化合物，这些化合物对于 2 dpf 和 5 dpf 的 TI 值均小于截止值但会诱导毒性表型的化合物，细化了测试浓度范围，并针对细化测试浓度的化合物将 TI 截止值从 ≥ 10 优化至 ≥ 3，这优化了斑马鱼发育测定的灵敏度达到 90.3%（28/31 阳性化合物在斑马鱼中具有致畸性）和 88.9% (40/45) 的总体可预测性。我们的研究结果强烈支持使用斑马鱼作为替代体内方法来筛选和评估候选药物的致畸性，以供监管机构接受。
The zebrafish as an alternative animal model for developmental toxicity testing has been extensively investigated, but its assay protocol was not harmonized yet. This study has validated and optimized the zebrafish developmental toxicity assay previously reported by multiple inter-laboratory studies in the United States and Europe. In this study, using this classical protocol, of 31 ICH-positive compounds, 23 compounds (74.2%) were teratogenic in zebrafish, five had false-negative results, and three were neither teratogenic nor non-teratogenic according to the protocol standard; of 14 ICH-negative compounds, 12 compounds (85.7%) were non-teratogenic in zebrafish and two had false-positive results. After we added an additional TI value in the zebrafish treated with testing compounds at 2 dpf along with the original 5 dpf, proposed a new category as the uncategorized compounds for those TI values smaller than the cutoff both at 2 dpf and 5 dpf but inducing toxic phenotypes, refined the testing concentration ranges, and optimized the TI cut-off value from ≥ 10 to ≥ 3 for compounds with refined testing concentrations, this optimized zebrafish developmental assay reached 90.3% sensitivity (28/31 positive compounds were teratogenic in zebrafish) and 88.9% (40/45) overall predictability. Our results from this study strongly support the use of zebrafish as an alternative in vivo method for screening and assessing the teratogenicity of candidate drugs for regulatory acceptance.
登录
查看更多内容
影响因子：
14.9
作者：
Kim S;Thiessen PA;Bolton EE;Chen J;Fu G;Gindulyte A;Han L;He J;He S;Shoemaker BA;Wang J;Yu B;Zhang J;Bryant SH
通讯作者：
Bryant SH
影响因子：
3.3
作者：
Gustafson, A. -L.;Stedman, D. B.;Augustine-Rauch, K. A.
通讯作者：
Augustine-Rauch, K. A.
影响因子：
3.1
作者：
Raghunath, Azhwar;Nagarajan, Raju;Perumal, Ekambaram
通讯作者：
Perumal, Ekambaram
影响因子：
4
作者：
Dooley, K;Zon, LI
通讯作者：
Zon, LI
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF