Mesens Natalie, Crawford Alexander D, Menke Aswin, Hung Pham Duc, Van Goethem Freddy, Nuyts Rik, Hansen Erik, Wolterbeek Andre, Van Gompel Jacky, De Witte Peter, Esguerra Camila V
Discovery Support and Investigative Toxicology group EU, Preclinical Development and Safety, Janssen Pharmaceutical Companies of Johnson & Johnson, Turnhoutseweg 30, 2340, Beerse, Belgium.
Laboratory for Molecular Biodiscovery, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven Campus Gasthuisberg, O&N 2, 08.4226 Herestraat 49, PB 824, 3000, Leuven, Belgium.
J Appl Toxicol. 2015 Sep;35(9):1017-29. doi: 10.1002/jat.3091. Epub 2015 Feb 6.
Drug-induced liver injury (DILI) is poorly predicted by single-cell-based assays, probably because of the lack of physiological interactions with other cells within the liver. An intact whole liver system such as one present in zebrafish larvae could provide added value in a screening strategy for DILI; however, the possible occurrence of other organ toxicities and the immature larval stage of the zebrafish might complicate accurate and fast analysis. We investigated whether expression analysis of liver-specific fatty acid binding protein 10a (lfabp10a) was an appropriate endpoint for assessing hepatotoxic effects in zebrafish larvae. It was found that expression analysis of lfabp10a was a valid marker, as after treatment with hepatotoxicants, dose-response curves could be obtained and statistically significant abnormal lfabp10 expression levels correlated with hepatocellular histopathological changes in the liver. However, toxicity in other vital organs such as the heart could impact liver outgrowth and thus had to be assessed concurrently. Whether zebrafish larvae were suitable for assessing human relevant drug-induced hepatotoxicity was assessed with hepatotoxicants and non-hepatotoxicants that have been marketed for human use and classified according to their mechanism of toxicity. The zebrafish larva showed promising predictivity towards a number of mechanisms and was capable of distinguishing between hepatotoxic and non-hepatotoxic chemical analogues, thus implying its applicability as a potential screening model for DILI.
基于单细胞的检测方法对药物性肝损伤(DILI)的预测效果不佳,这可能是因为缺乏与肝脏内其他细胞的生理相互作用。完整的全肝系统,如斑马鱼幼体中存在的系统,在DILI的筛选策略中可能会提供额外价值;然而,其他器官毒性的可能发生以及斑马鱼幼体阶段的不成熟可能会使准确快速的分析变得复杂。我们研究了肝脏特异性脂肪酸结合蛋白10a(lfabp10a)的表达分析是否是评估斑马鱼幼体肝毒性作用的合适终点。结果发现,lfabp10a的表达分析是一个有效的标志物,因为在用肝毒性药物处理后,可以获得剂量反应曲线,并且lfabp10表达水平的统计学显著异常与肝脏中的肝细胞组织病理学变化相关。然而,心脏等其他重要器官的毒性可能会影响肝脏的生长,因此必须同时进行评估。我们用已上市用于人类的肝毒性药物和非肝毒性药物,并根据其毒性机制进行分类,评估斑马鱼幼体是否适合评估人类相关的药物性肝毒性。斑马鱼幼体对多种机制显示出有前景的预测能力,并且能够区分肝毒性和非肝毒性化学类似物,这意味着它作为DILI潜在筛选模型的适用性。