Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School and Shriners Hospitals for Children, Boston, MA, 02114, USA.
Department of Biomedical Engineering, Rutgers University, Piscataway, NJ, 08854, USA.
Sci Rep. 2019 Nov 1;9(1):15848. doi: 10.1038/s41598-019-52174-w.
CYP3A4, a cytochrome P450 enzyme regulated by the nuclear receptor PXR, is involved in most of the drug metabolizing pathways. Studying the regulation/induction of CYP3A4 and PXR is critical in toxicology and drug-drug interaction (DDI) studies. Primary human hepatocytes constitute the preferred in vitro platform for drug development efforts. However, they are expensive, scarce and heterogeneous. Hepatic cell lines, such as Huh7, could provide a cost-effective alternative, however, they express negligible amounts of CYP450s and PXR. In this study, we show that dinaciclib, a potent cyclin dependent kinase inhibitor, significantly increases the basal CYP3A4 and PXR levels in 24 hours. We also demonstrated that matured Huh7s can be used for drug induction studies, where CYP3A4, CYP1A2, CYP2C9, and CYP2C19 inductions were achieved following rifampicin treatment. More importantly, through a direct demonstration using amiodarone and rifampicin as model drugs, we showed that matured Huh7s present a suitable platform for DDI studies.
CYP3A4 是一种细胞色素 P450 酶，受核受体 PXR 调控，参与大多数药物代谢途径。研究 CYP3A4 和 PXR 的调控/诱导对于毒理学和药物相互作用（DDI）研究至关重要。原代人肝细胞是药物开发的首选体外平台。然而，它们昂贵、稀缺且异质。Huh7 等肝细胞系可以提供一种具有成本效益的替代方案，但它们表达的 CYP450 和 PXR 数量可以忽略不计。在这项研究中，我们表明，强效细胞周期蛋白依赖性激酶抑制剂达昔单抗在 24 小时内显著增加基础 CYP3A4 和 PXR 水平。我们还证明，成熟的 Huh7 可用于药物诱导研究，在利福平处理后可实现 CYP3A4、CYP1A2、CYP2C9 和 CYP2C19 的诱导。更重要的是，通过使用胺碘酮和利福平作为模型药物的直接证明，我们表明成熟的 Huh7 是进行 DDI 研究的合适平台。
