title: Bcrp1;Mdr1a/b;Mrp2 combination knockout mice: altered disposition of the dietary carcinogen PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) and its genotoxic metabolites.
abstract: The multidrug transporters breast cancer resistance protein (BCRP), multidrug-resistance protein 1 (MDR1), and multidrug-resistance-associated protein (MRP) 2 and 3 eliminate toxic compounds from tissues and the body and affect the pharmacokinetics of many drugs and other potentially toxic compounds. The food-derived carcinogen PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) is transported by BCRP, MDR1, and MRP2. To investigate the overlapping functions of Bcrp1, Mdr1a/b, and Mrp2 in vivo, we generated Bcrp1;Mdr1a/b;Mrp2(-/-) mice, which are viable and fertile. These mice, together with Bcrp1;Mrp2;Mrp3(-/-) mice, were used to study the effects of the multidrug transporters on the pharmacokinetics of PhIP and its metabolites. Thirty minutes after oral or intravenous administration of PhIP (1 mg/kg), the PhIP levels in the small intestine were reduced 4- to 6-fold in Bcrp1;Mdr1a/b;Mrp2(-/) (-) and Bcrp1;Mrp2;Mrp3(-/-) mice compared with wild-type mice. Fecal excretion of PhIP was reduced 8- to 20-fold in knockouts. Biliary PhIP excretion was reduced 41-fold in Bcrp1;Mdr1a/b;Mrp2(-/-) mice. Biliary and small intestine levels of PhIP metabolites were reduced in Bcrp1;Mrp2-deficient mice. Furthermore, in both knockout strains, kidney levels and urinary excretion of genotoxic PhIP-metabolites were significantly increased, suggesting that reduced biliary excretion of PhIP and PhIP metabolites leads to increased urinary excretion of these metabolites and increased systemic exposure. Bcrp1 and Mdr1a limited PhIP brain accumulation. In Bcrp1;Mrp2;Mrp3(-/-), but not Bcrp1;Mdr1a/b;Mrp(-/-) mice, the carcinogenic metabolites N2-OH-PhIP (2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine) and PhIP-5-sulfate (a genotoxicity marker) accumulated in liver tissue, indicating that Mrp3 is involved in the sinusoidal secretion of these compounds. We conclude that Bcrp1, Mdr1a/b, Mrp2, and Mrp3 significantly affect tissue disposition and biliary and fecal elimination of PhIP and its carcinogenic metabolites and may affect PhIP-induced carcinogenesis as a result.
authors: Divisions of Molecular Oncology (M.L.H.V., E.v.d.S., A.v.E., E.W., A.H.S.) and Clinical Pharmacology (J.H.M.S.), The Netherlands Cancer Institute, Amsterdam, The Netherlands; Division of Pharmacy & Pharmacology, Slotervaart Hospital, Amsterdam, The Netherlands (S.F.T., H.R., J.H.B.); Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands (L.B., T.F.A.d.G.); and Department of Pharmaceutical Sciences, Science Faculty, Utrecht University, Utrecht, The Netherlands (J.H.M.S., J.H.B.).
authors: Maria L H
authors: ML
authors: Vlaming
authors: Sebastiaan F
authors: SF
authors: Teunissen
authors: Evita
authors: E
authors: van de Steeg
authors: Anita
authors: A
authors: van Esch
authors: Els
authors: E
authors: Wagenaar
authors: Luc
authors: L
authors: Brunsveld
authors: Tom F A
authors: TF
authors: de Greef
authors: Hilde
authors: H
authors: Rosing
authors: Jan H M
authors: JH
authors: Schellens
authors: Jos H
authors: JH
authors: Beijnen
authors: Alfred H
authors: AH
authors: Schinkel
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics* / Q000378:metabolism; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism; D042501:Angiogenic Proteins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002273:Carcinogens / Q000378:metabolism* / Q000493:pharmacokinetics; D007093:Imidazoles / Q000378:metabolism* / Q000493:pharmacokinetics; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D000097202:ATP-Binding Cassette Sub-Family B Member 4
chemical_list: D018435:ATP Binding Cassette Transporter, Subfamily B; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D042501:Angiogenic Proteins; D002273:Carcinogens; D007093:Imidazoles; D000097202:ATP-Binding Cassette Sub-Family B Member 4; C501412:Abcg2 protein, mouse; C543400:mitogen-regulated protein 3, mouse; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine; C095122:multidrug resistance protein 3