Drug-induced liver injury (DILI) is a life-threatening, adverse reaction to certain drugs. The onset and extent of DILI can vary drastically in different patients using identical drugs. Association studies suggested that subtle differences in DNA methylation may help explain the individual differences in DILI. However, there are very few experimental methods to confirm such associations. In this study, we established a novel DNA methylation functional detection system in human hepatocytes, using CRISPR/dCas9 for targeted modification of DNA methylation, and set four parameters to indicate the liver injury by cell model. Using this system, we validated the association of hypermethylation of CYP2D6 and CYP2E1 with rifampin-induced DILI. Our results revealed that, following treatment of HepaRG cells with rifampin, the methylation levels of CYP2D6 and CYP2E1 were inversely proportional to cell viability and glutathione content, and directly proportional to caspase 3/7 activity. We expect that our methylation detection system will serve as a useful tool in validating correlations between DNA methylation and DILI in other in vitro systems. Our results establish a foundation for future investigations to better understand the mechanisms underlying DILI and may aid in advancing personalized DILI medicine.

中文翻译：

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甲基化功能检测肝细胞系统验证了DNA甲基化与药物诱导的肝损伤之间的相关性。

药物性肝损伤（DILI）是对某些药物的危及生命的不良反应。在使用相同药物的不同患者中，DILI的发作和程度可能会发生巨大变化。协会研究表明，DNA甲基化的细微差异可能有助于解释DILI的个体差异。但是，很少有实验方法可以证实这种关联。在这项研究中，我们建立了一种新的人类肝细胞DNA甲基化功能检测系统，使用CRISPR / dCas9对DNA甲基化进行靶向修饰，并设置了四个参数来通过细胞模型指示肝损伤。使用该系统，我们验证了CYP2D6和CYP2E1的甲基化与利福平诱导的DILI的相关性。我们的结果表明，在用利福平处理HepaRG细胞后，CYP2D6和CYP2E1的甲基化水平与细胞活力和谷胱甘肽含量成反比，与caspase 3/7活性成正比。我们希望我们的甲基化检测系统将成为验证其他体外系统中DNA甲基化与DILI之间相关性的有用工具。我们的结果为将来的研究奠定了基础，以更好地了解DILI的潜在机制，并可能有助于推进DILI个性化医学的发展。