title: Solute Carrier Family of the Organic Anion-Transporting Polypeptides 1A2- Madin-Darby Canine Kidney II: A Promising In Vitro System to Understand the Role of Organic Anion-Transporting Polypeptide 1A2 in Blood-Brain Barrier Drug Penetration.
abstract: Organic anion-transporting polypeptide (OATP) 1A2 has the potential to be a target for central nervous system drug delivery due to its luminal localization at the human blood-brain barrier and broad substrate specificity. We found OATP1A2 mRNA expression in the human brain to be comparable to breast cancer resistance protein and OATP2B1 and much higher than P-glycoprotein (P-gp), and confirmed greater expression in the brain relative to other tissues. The goal of this study was to establish a model system to explore OATP1A2-mediated transcellular transport of substrate drugs and the interplay with P-gp. In vitro (human embryonic kidney 293 cells stably expressing Oatp1a4, the closest murine isoform) and in vivo (naïve and Oatp1a4 knock-out mice) studies with OATP1A2 substrate triptan drugs demonstrated that these drugs were not Oatp1a4 substrates. This species difference demonstrates that the rodent is not a good model to investigate the active brain uptake of potential OATP1A2 substrates. Thus, we constructed a novel OATP1A2 expressing Madin-Darby canine kidney (MDCK) II wild type and an MDCKII-multidrug resistance protein 1 (MDR1) system using BacMam virus transduction. The spatial expression pattern of OATP1A2 after transduction in MDCKII-MDR1 cells was superimposed to P-gp, confirming apical membrane localization. OATP1A2-mediated uptake of zolmitriptan, rosuvastatin, and fexofenadine across monolayers increased with increasing OATP1A2 protein expression. OATP1A2 counteracted P-gp efflux for cosubstrates zolmitriptan and fexofenadine. A three-compartment model incorporating OATP1A2-mediated influx was used to quantitatively describe the time- and concentration-dependent apical-to-basolateral transcellular transport of rosuvastatin across OATP1A2 expressing the MDCKII monolayer. This novel, simple and versatile experimental system is useful for understanding the contribution of OATP1A2-mediated transcellular transport across barriers, such as the blood-brain barrier.
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Houfu
authors.initials: H
authors.lastname: Liu
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Na
authors.initials: N
authors.lastname: Yu
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Sijie
authors.initials: S
authors.lastname: Lu
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Sumito
authors.initials: S
authors.lastname: Ito
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Bhagwat
authors.initials: B
authors.lastname: Prasad
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Enuo
authors.initials: E
authors.lastname: He
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Xinyan
authors.initials: X
authors.lastname: Lu
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Han
authors.initials: H
authors.lastname: Xu
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Gang
authors.initials: G
authors.lastname: An
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Jashvant D
authors.initials: JD
authors.lastname: Unadkat
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kusuhara
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.).
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Sugiyama
authors.affiliation: Drug Metabolism and Pharmacokinetics (H.L., N.Y., S.L., X.L., Y.L., F.W., J.S.) and Molecular Discovery Research (H.X., G.A.), Platform Technology and Science, GlaxoSmithKline Research and Development, Shanghai, China; Modelling and Translational Biology, Platform Technology and Science, GlaxoSmithKline, Ware, United Kingdom (E.H.); Department of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, University Of Tokyo, Tokyo, Japan (S.I., X.Z., H.K.); Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, Kanagawa, Japan (Y.S.); and Department of Pharmaceutics, University of Washington, Seattle, Washington (B.P., J.D.U.) jasminder.j.sahi@gsk.com.
authors.forename: Jasminder
authors.initials: J
authors.lastname: Sahi
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000818:Animals; D001693:Biological Transport, Active; D001812:Blood-Brain Barrier / Q000378:metabolism*; D004285:Dogs; D057809:HEK293 Cells; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D018345:Mice, Knockout; D008954:Models, Biological; D027361:Organic Anion Transporters / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D000068718:Rosuvastatin Calcium / Q000493:pharmacokinetics; D013045:Species Specificity; D014018:Tissue Distribution; D014363:Tryptamines / Q000378:metabolism
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D027361:Organic Anion Transporters; D004364:Pharmaceutical Preparations; C512988:SLCO1A2 protein, human; D014363:Tryptamines; D000068718:Rosuvastatin Calcium