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Expanding the Efflux In Vitro Assay Toolbox: A CRISPR-Cas9 Edited MDCK Cell Line with Human BCRP and Completely Lacking Canine MDR1
- Source :
- Journal of Pharmaceutical Sciences. 110:388-396
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- 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 (MDCKcMDR1-KO) for stable expression of human BCRP (hBCRP) with no endogenous canine MDR1 (cMDR1) expression (MDCK-hBCRPcMDR1-KO). 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 MDCKcMDR1-KO and a previously established, corresponding cell line overexpressing human MDR1 (hMDR1), MDCK-hMDR1cMDR1-KO. Functional studies with MDCK-hBCRPcMDR1-KO confirmed high hBCRP activity. The MDCK-hBCRPcMDR1-KO cell line together with the MDCK-hMDR1cMDR1-KO 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.
- Subjects :
- Proteomics
Abcg2
Quantitative proteomics
Pharmaceutical Science
ATP-binding cassette (ABC) transporter(s)
02 engineering and technology
030226 pharmacology & pharmacy
Permeability
Cell Line
Madin Darby Canine Kidney Cells
Pharmaceutical Sciences
03 medical and health sciences
Dogs
0302 clinical medicine
Multidrug Resistance Protein 1
Efflux pumps
ATP Binding Cassette Transporter, Subfamily G, Member 2
Animals
Humans
ATP Binding Cassette Transporter, Subfamily B, Member 1
P-glycoprotein (P-gp)
Membrane transporter
biology
Chemistry
MDCK cells
Drug transport
Biological Transport
Farmaceutiska vetenskaper
021001 nanoscience & nanotechnology
Neoplasm Proteins
Cell biology
Drug development
Cell culture
Proteome
biology.protein
Breast cancer resistance protein (BCRP)
Efflux
CRISPR-Cas Systems
0210 nano-technology
Subjects
Details
- ISSN :
- 00223549
- Volume :
- 110
- Database :
- OpenAIRE
- Journal :
- Journal of Pharmaceutical Sciences
- Accession number :
- edsair.doi.dedup.....03d5bae62eb3a6ba326e577733489bec