Department of Pharmacy, Drug Delivery Research Group, Uppsala University, Uppsala, Sweden; Department of Pharmacy, Uppsala University Drug Optimization and Pharmaceutical Profiling Platform (UDOPP), Uppsala, Sweden.
Department of Pharmacy, Drug Delivery Research Group, Uppsala University, Uppsala, Sweden.
J Pharm Sci. 2021 Jan;110(1):388-396. doi: 10.1016/j.xphs.2020.09.039. Epub 2020 Sep 29.
The Breast Cancer Resistance Protein (BCRP) is a key transporter in drug efflux and drug-drug interactions. However, endogenous expression of Multidrug Resistance Protein 1 (MDR1) confounds the interpretation of BCRP-mediated transport in in vitro models. Here we used a CRISPR-Cas9 edited Madin-Darby canine kidney (MDCK) II cell line (MDCK) for stable expression of human BCRP (hBCRP) with no endogenous canine MDR1 (cMDR1) expression (MDCK-hBCRP). Targeted quantitative proteomics verified expression of hBCRP, and global analysis of the entire proteome corroborated no or very low background expression of other drug transport proteins or metabolizing enzymes. This new cell line, had similar proteome like MDCK and a previously established, corresponding cell line overexpressing human MDR1 (hMDR1), MDCK-hMDR1. Functional studies with MDCK-hBCRP confirmed high hBCRP activity. The MDCK-hBCRP cell line together with the MDCK-hMDR1 easily and accurately identified shared or specific substrates of the hBCRP and the hMDR1 transporters. These cell lines offer new, improved in vitro tools for the assessment of drug efflux and drug-drug interactions in drug development.
乳腺癌耐药蛋白 (BCRP) 是药物外排和药物相互作用的关键转运体。然而，多药耐药蛋白 1 (MDR1) 的内源性表达使 BCRP 介导的转运在体外模型中的解释变得复杂。在这里，我们使用 CRISPR-Cas9 编辑的犬肾细胞 (MDCK) II 细胞系 (MDCK) 稳定表达人 BCRP (hBCRP)，而没有内源性犬 MDR1 (cMDR1) 表达 (MDCK-hBCRP)。靶向定量蛋白质组学验证了 hBCRP 的表达，对整个蛋白质组的全面分析证实了其他药物转运蛋白或代谢酶的表达水平非常低或没有。这种新的细胞系与 MDCK 具有相似的蛋白质组，并且与先前建立的、过表达人 MDR1 (hMDR1) 的相应细胞系 MDCK-hMDR1 相似。使用 MDCK-hBCRP 的功能研究证实了 hBCRP 的高活性。MDCK-hBCRP 细胞系与 MDCK-hMDR1 一起，可轻松准确地鉴定 hBCRP 和 hMDR1 转运体的共同或特定底物。这些细胞系为药物开发中药物外排和药物相互作用的评估提供了新的、改进的体外工具。
