title: A Human Renal Proximal Tubule Cell Line with Stable Organic Anion Transporter 1 and 3 Expression Predictive for Antiviral-Induced Toxicity.
abstract: Drug-induced nephrotoxicity still hampers drug development, because current translation from in vitro or animal studies to human lacks high predictivity. Often, renal adverse effects are recognized only during clinical stages of drug development. The current study aimed to establish a robust and a more complete human cell model suitable for screening of drug-related interactions and nephrotoxicity. In addition to endogenously expressed renal organic cation transporters and efflux transporters, conditionally immortalized proximal tubule epithelial cells (ciPTEC) were completed by transduction of cells with the organic anion transporter (OAT) 1 or OAT3. Fluorescence-activated cell sorting upon exposure to the OAT substrate fluorescein successfully enriched transduced cells. A panel of organic anions was screened for drug-interactions in ciPTEC-OAT1 and ciPTEC-OAT3. The cytotoxic response to the drug-interactions with antivirals was further examined by cell viability assays. Upon subcloning, concentration-dependent fluorescein uptake was found with a higher affinity for ciPTEC-OAT1 (Km = 0.8 ± 0.1 μM) than ciPTEC-OAT3 (Km = 3.7 ± 0.5 μM). Co-exposure to known OAT1 and/or OAT3 substrates (viz. para-aminohippurate, estrone sulfate, probenecid, furosemide, diclofenac, and cimetidine) in cultures spanning 29 passage numbers revealed relevant inhibitory potencies, confirming the robustness of our model for drug-drug interactions studies. Functional OAT1 was directly responsible for cytotoxicity of adefovir, cidofovir, and tenofovir, while a drug interaction with zidovudine was not associated with decreased cell viability. Our data demonstrate that human-derived ciPTEC-OAT1 and ciPTEC-OAT3 are promising platforms for highly predictive drug screening during early phases of drug development.
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.
authors.forename: Tom T G
authors.initials: TT
authors.lastname: Nieskens
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.
authors.forename: Janny G P
authors.initials: JG
authors.lastname: Peters
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.
authors.forename: Marieke J
authors.initials: MJ
authors.lastname: Schreurs
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: Smits
authors.affiliation: Department of Laboratory Medicine - Laboratory of Hematology, Radboud University Medical Centre, Nijmegen, The Netherlands.
authors.forename: Rob
authors.initials: R
authors.lastname: Woestenenk
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.
authors.forename: Katja
authors.initials: K
authors.lastname: Jansen
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.
authors.forename: Thom K
authors.initials: TK
authors.lastname: van der Made
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.
authors.forename: Melanie
authors.initials: M
authors.lastname: Röring
authors.affiliation: Innovative Medicines, Drug Safety and Metabolism, AstraZeneca R&D, Mölndal, Sweden.
authors.forename: Constanze
authors.initials: C
authors.lastname: Hilgendorf
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands. martijn.wilmer@radboudumc.nl.|Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Radboud University medical centre, P.O. box 9101, 6500 HB, Nijmegen, The Netherlands. martijn.wilmer@radboudumc.nl.
authors.forename: Martijn J
authors.initials: MJ
authors.lastname: Wilmer
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute of Molecular Life Sciences, Radboud University medical center, Nijmegen, The Netherlands.|Division Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Rosalinde
authors.initials: R
authors.lastname: Masereeuw
mesh_terms: D016475:3T3 Cells; D000225:Adenine / Q000031:analogs & derivatives / Q000633:toxicity; D000818:Animals; D000998:Antiviral Agents / Q000633:toxicity*; D002460:Cell Line; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D000077404:Cidofovir; D003596:Cytosine / Q000031:analogs & derivatives / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D005544:Forecasting; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D027362:Organic Anion Transport Protein 1 / Q000096:biosynthesis*; D029362:Organic Anion Transporters, Sodium-Independent / Q000096:biosynthesis*; D063065:Organophosphonates / Q000633:toxicity
chemical_list: D000998:Antiviral Agents; D027362:Organic Anion Transport Protein 1; D029362:Organic Anion Transporters, Sodium-Independent; D063065:Organophosphonates; C114338:organic anion transport protein 3; C053001:adefovir; D003596:Cytosine; D000225:Adenine; D000077404:Cidofovir
keywords: antivirals; drug-drug interactions; nephrotoxicity; organic anion transport; proximal tubule epithelial cell
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