Unidad de Hepatología Experimental, Torre A. Instituto Investigación Sanitaria La Fe, Av Fernando Abril Martorell 106, 46026, Valencia, Spain.
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, 46010, Valencia, Spain.
Arch Toxicol. 2019 Feb;93(2):519-532. doi: 10.1007/s00204-018-2349-y. Epub 2018 Nov 13.
Drug-induced liver injury (DILI) constitutes one of the most frequent reasons of restricted-use warnings as well as withdrawals of drugs in postmarketing and poses an important concern for the pharmaceutical industry. The current hepatic in vivo and in vitro models for DILI detection have shown clear limitations, mainly for studies of long-term hepatotoxicity. For this reason, we here evaluated the potential of using Upcytes human hepatocytes (UHH) for repeated-dose long-term exposure to drugs. The UHH were incubated with 15 toxic and non-toxic compounds for up to 21 days using a repeated-dose approach, and, in addition to conventional examination of effects on viability, the mechanisms implicated in cell toxicity were also assessed by means of high-content screening. The UHH maintained the expression and activity levels of drug-metabolizing enzymes for up to 21 days of culture and became more sensitive to the toxic compounds after extended exposures, showing inter-donor differences which would reflect variability among the population. The assay also allowed to detect the main mechanisms implicated in the toxicity of each drug as well as identifying special susceptibilities depending on the donor. UHH can be used for a long-term repeated detection of DILI at clinically relevant concentrations and also offers key mechanistic features of drug-induced hepatotoxicity. This system is therefore a promising tool in preclinical testing of human relevance that could help to reduce and/or replace animal testing for drug adverse effects.
药物性肝损伤（DILI）是导致药品在上市后受到限制使用警告甚至撤市的最常见原因之一，也是制药行业关注的重要问题。目前用于检测 DILI 的体内和体外肝模型显示出明显的局限性，主要是因为它们无法研究长期肝毒性。出于这个原因，我们在此评估了使用 Upcytes 人原代肝细胞（UHH）进行药物重复剂量长期暴露的潜力。UHH 采用重复剂量方法与 15 种有毒和无毒化合物孵育长达 21 天，除了常规的细胞活力检测外，还通过高内涵筛选评估了与细胞毒性相关的机制。UHH 在培养 21 天内保持了药物代谢酶的表达和活性水平，并且在延长暴露后对有毒化合物变得更加敏感，表现出供体间的差异，这将反映人群中的变异性。该检测还可以检测到每种药物毒性涉及的主要机制，并根据供体确定特殊的易感性。UHH 可用于在临床相关浓度下进行长期重复的 DILI 检测，并且还提供了药物诱导肝毒性的关键机制特征。因此，该系统是一种很有前途的临床前人类相关性测试工具，可以帮助减少和/或替代药物不良反应的动物测试。
