Division of the National Toxicology Program, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA.
Neurocrine Biosciences Inc., San Diego, California, 92130, USA.
Toxicol Sci. 2022 Jul 28;188(2):198-207. doi: 10.1093/toxsci/kfac053.
Compound toxicity data obtained from independent zebrafish laboratories can vary vastly, complicating the use of zebrafish screening for regulatory decisions. Differences in the assay protocol parameters are the primary source of variability. We investigated this issue by utilizing data from the NTP DNT-DIVER database (https://doi.org/10.22427/NTP-DATA-002-00062-0001-0000-1, last accessed June 2, 2022), which consists of data from zebrafish developmental toxicity (devtox) and locomotor response (designated as "neurotox") screens from 3 independent laboratories, using the same set of 87 compounds. The data were analyzed using the benchmark concentration (BMC) modeling approach, which estimates the concentration of interest based on a predetermined response threshold. We compared the BMC results from 3 laboratories (A, B, C) in 3 toxicity outcome categories: mortality, cumulative devtox, and neurotox, in terms of activity calls and potency values. We found that for devtox screening, laboratories with similar/same protocol parameters (B vs C) had an active call concordance as high as 86% with negligible potency difference. For neurotox screening, active call concordances between paired laboratories are lower than devtox screening (highest 68%). When protocols with different protocol parameters were compared, the concordance dropped, and the potency shift was on average about 3.8-fold for the cumulative devtox outcome and 5.8-fold for the neurotox outcome. The potential contributing protocol parameters for potency shift are listed or ranked. This study provides a quantitative assessment of the source of variability in zebrafish screening protocols and sets the groundwork for the ongoing Systematic Evaluation of the Application of Zebrafish in Toxicology effort at the National Toxicology Program.
来自独立的斑马鱼实验室的复合毒性数据差异很大，这使得斑马鱼筛选在监管决策中的应用变得复杂。测定方案参数的差异是变异性的主要来源。我们通过利用 NTP DNT-DIVER 数据库（https://doi.org/10.22427/NTP-DATA-002-00062-0001-0000-1，最后访问日期为 2022 年 6 月 2 日）中的数据来研究这个问题，该数据库由来自 3 个独立实验室的斑马鱼发育毒性（devtox）和运动反应（指定为“神经毒性”）筛选数据组成，使用相同的 87 种化合物。数据使用基准浓度（BMC）建模方法进行分析，该方法根据预定的反应阈值估算感兴趣的浓度。我们根据活性呼叫和效价值，将来自 3 个实验室（A、B 和 C）的 3 种毒性结果类别（死亡率、累积 devtox 和神经毒性）的 BMC 结果进行比较。我们发现，对于 devtox 筛选，具有相似/相同方案参数的实验室（B 与 C）的活性呼叫一致性高达 86%，效价差异可忽略不计。对于神经毒性筛选，配对实验室之间的活性呼叫一致性低于 devtox 筛选（最高为 68%）。当比较具有不同方案参数的方案时，一致性下降，效价变化平均为累积 devtox 结果的 3.8 倍，为神经毒性结果的 5.8 倍。列出或排名了导致效价变化的潜在协议参数。这项研究定量评估了斑马鱼筛选方案中的变异性来源，并为国家毒理学计划中正在进行的毒理学中应用斑马鱼的系统评估奠定了基础。
