Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa 920-1192, Japan.
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa 920-1192, Japan; Department of Drug Safety Sciences, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan.
Toxicol Lett. 2014 Jul 3;228(1):13-24. doi: 10.1016/j.toxlet.2014.04.005. Epub 2014 Apr 15.
Drug-induced liver injury (DILI) is a major safety concern in drug development and clinical pharmacotherapy. However, prediction of DILI is difficult because the underlying mechanisms are not fully understood. To establish a novel cell-based screening system to suggest drugs with hepatotoxic potential in preclinical drug development, comprehensive gene expression analyses during in vivo DILI are necessary. Using in vivo mouse DILI models and 4 sets of hepatotoxic positive and non-hepatotoxic drugs, we found that the hepatic mRNA levels of S100A8; S100A9; "NATCH, LRR, and pyrin domain-containing protein 3" (NALP3); interleukin (IL)-1β; and the receptor for advanced glycation endproducts (RAGE) were commonly increased in hepatotoxic drug-administered mice compared to non-hepatotoxic drug-administered mice. To clarify whether these 5 in vivo biomarkers can be applied to a cell-based screening system, we adapted human liver microsomes (HLM) in the presence of NADPH to assess the metabolic activation reaction, and we also adapted human monocytic leukemia cells HL-60, K562, KG-1 and THP-1 to assess the effects on mRNA expression of immune- and inflammatory-related factors. We investigated 30 clinical drugs with different safety profiles with regard to DILI and found that the total sum score of gene expression levels of S100A8, S100A9, RAGE, NALP3 and IL-1β mRNA in HL-60 or K562 cells incubated with HLM, could identify drugs at high risk for hepatotoxicity. We proposed the use of the total sum score of gene expression level for assessing metabolic activation by drug-metabolizing enzymes and immune- and inflammatory-related factors for the risk assessment of DILI in preclinical drug development.
药物性肝损伤（DILI）是药物开发和临床药物治疗中的一个主要安全问题。然而，由于其潜在机制尚未完全阐明，因此预测 DILI 具有一定难度。为了建立一种新的基于细胞的筛选系统，以在临床前药物开发中提示具有潜在肝毒性的药物，有必要对体内 DILI 过程中的全面基因表达进行分析。本研究使用体内小鼠 DILI 模型和 4 组肝毒性阳性和非肝毒性药物，发现与非肝毒性药物处理的小鼠相比，肝毒性药物处理的小鼠的肝脏 mRNA 水平普遍升高，这些基因包括 S100A8、S100A9、“NATCH、LRR 和吡喃结构域蛋白 3”（NALP3）、白细胞介素（IL）-1β 和晚期糖基化终产物受体（RAGE）。为了明确这 5 个体内生物标志物是否可应用于基于细胞的筛选系统，我们在存在 NADPH 的情况下适应人肝微粒体（HLM）来评估代谢激活反应，并且我们还适应人单核白血病细胞 HL-60、K562、KG-1 和 THP-1 来评估它们对免疫和炎症相关因子的 mRNA 表达的影响。我们研究了 30 种具有不同 DILI 安全性特征的临床药物，发现与 HLM 孵育的 HL-60 或 K562 细胞中 S100A8、S100A9、RAGE、NALP3 和 IL-1β mRNA 的基因表达水平的总得分，可识别出高风险肝毒性的药物。我们提出使用药物代谢酶和免疫及炎症相关因子的基因表达水平的总得分来评估代谢激活，用于临床前药物开发中 DILI 的风险评估。
