National Center for Computational Toxicology, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC, USA.
National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC, USA.
Toxicol Appl Pharmacol. 2018 Sep 1;354:24-39. doi: 10.1016/j.taap.2018.04.001. Epub 2018 Apr 5.
Medium- to high-throughput in vitro assays that recapitulate the critical processes of nervous system development have been proposed as a means to facilitate rapid testing and identification of chemicals which may affect brain development. In vivo neurodevelopment is a complex progression of distinct cellular processes. Therefore, batteries of in vitro assays that model and quantify effects on a variety of neurodevelopmental processes have the potential to identify chemicals which may affect brain development at different developmental stages. In the present study, the results of concentration-response screening of 67 reference chemicals in a battery of high content imaging and microplate reader-based assays that evaluate neural progenitor cell proliferation, neural proginitor cell apoptosis, neurite initiation/outgrowth, neurite maturation and synaptogenesis are summarized and compared. The assay battery had a high degree of combined sensitivity (87%) for categorizing chemicals known to affect neurodevelopment as active and a moderate degree of combined specificity (71%) for categorizing chemicals not associated with affects on neurodevelopment as inactive. The combined sensitivity of the assay battery was higher compared to any individual assay while the combined specificity of the assay battery was lower compared to any individual assay. When selectivity of effects for a neurodevelopmental endpoint as compared to general cytotoxicity was taken into account, the combined sensitivity of the assay battery decreased (68%) while the combined specificity increased (93%). The identity and potency of chemicals identified as active varied across the assay battery, underscoring the need for use of a combination of diverse in vitro models to comprehensively screen chemicals and identify those which potentially affect neurodevelopment. Overall, these data indicate that a battery of assays which address many different processes in nervous system development may be used to identify potential developmental neurotoxicants and to distinguish specific from generalized cytotoxic effects with a high degree of success.
中高通量的体外检测方法可以重现神经系统发育的关键过程，这被提议作为一种快速测试和识别可能影响大脑发育的化学物质的方法。体内神经发育是一个复杂的细胞过程。因此，有一系列可以模拟和量化对各种神经发育过程的影响的体外检测方法，有可能识别出在不同发育阶段可能影响大脑发育的化学物质。在本研究中，总结和比较了 67 种参考化学物质在一系列高内涵成像和微孔板阅读器检测方法中的浓度反应筛选结果，这些方法评估神经祖细胞增殖、神经祖细胞凋亡、神经突起始/生长、神经突成熟和突触发生。该检测试剂盒对已知影响神经发育的化学物质的分类具有很高的综合敏感性（87%），对与神经发育无关联的化学物质的分类具有中等程度的综合特异性（71%）。与任何单一检测方法相比，该检测试剂盒的综合敏感性更高，而与任何单一检测方法相比，该检测试剂盒的综合特异性更低。当考虑到对神经发育终点的效应选择性与一般细胞毒性时，检测试剂盒的综合敏感性降低（68%），而综合特异性增加（93%）。被鉴定为活性的化学物质的身份和效力在整个检测试剂盒中有所不同，这强调了需要使用多种不同的体外模型来全面筛选化学物质，并识别那些可能影响神经发育的化学物质。总的来说，这些数据表明，一组针对神经系统发育中许多不同过程的检测方法可以用于识别潜在的发育神经毒物，并以很高的成功率区分特定的和普遍的细胞毒性效应。
