title: Endogenous drug transporters in in vitro and in vivo models for the prediction of drug disposition in man.
abstract: The epithelial canine and porcine kidney cell lines MDCK, MDCKII and LLC-PK1, respectively are employed to establish recombinant models of drug transport. Endogenous drug carriers in these cells may contribute to the activities of recombinant drug transporters, thus making it difficult to assess their properties. We analysed the expression of endogenous transporters in these cell lines by RT-PCR and by determining drug transporter activities. Concerning drug efflux, multidrug resistance protein 1 (MDR1) and MRP1 mRNAs were found in all lines. MRP2 mRNA was expressed in all cell lines except MDCK. Transepithelial transport of vinblastine and its modulation by a MDR1-specific inhibitor or by the MDR1- and MRP-inhibitor verapamil, indicated that MDCKII cells have, in comparisons to the other cell lines, relatively high levels of functional MDR1 while vinblastine transport in MDCK cells is likely to be mediated more by MRP1. Notably, LLC-PK1 cells displayed little activity attributable to either MDR1 and MRP1, thus making them suitable for the expression of these efflux pumps. Of the drug uptake carriers, OATP-A mRNA was only expressed in MDCK cells. OATP-C mRNA was barely detectable in MDCK cells and absent in MDCKII and LLC-PK1 cells. In agreement with transcriptional profiling, the OATP-mediated uptake of either estradiol-glucuronide or estrone-sulfate was either absent or barely detectable in all cell lines thus implying that they are suitable to establish recombinant models for human OATP's. Transcriptional profiling was also performed on porcine and canine tissues and revealed that MRP1 was expressed in canine but not in human or porcine liver, whereas surprisingly OATP-C was expressed in canine kidney but only in human and porcine liver. The findings presented are relevant to the use of porcine and canine models for drug disposition.
authors.affiliation: Biomedical Research Centre, University of Dundee, Ninewells Hospital and Medical School, DD1 9SY, Dundee, UK.
authors.forename: Lay Beng
authors.initials: LB
authors.lastname: Goh
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Spears
authors.forename: Denggao
authors.initials: D
authors.lastname: Yao
authors.forename: Andy
authors.initials: A
authors.lastname: Ayrton
authors.forename: Paul
authors.initials: P
authors.lastname: Morgan
authors.forename: C
authors.initials: C
authors.lastname: Roland Wolf
authors.forename: Thomas
authors.initials: T
authors.lastname: Friedberg
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D018076:DNA, Complementary / Q000032:analysis; D004285:Dogs; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D027381:Liver-Specific Organic Anion Transporter 1 / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D008954:Models, Biological; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011237:Predictive Value of Tests; D013552:Swine
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018076:DNA, Complementary; D027381:Liver-Specific Organic Anion Transporter 1; D004364:Pharmaceutical Preparations