title: Application of RPTEC/TERT1 cells for investigation of repeat dose nephrotoxicity: A transcriptomic study.
abstract: The kidney is a major target organ for toxicity. Incidence of chronic kidney disease (CKD) is increasing at an alarming rate due to factors such as increasing population age and increased prevalence of heart disease and diabetes. There is a major effort ongoing to develop superior predictive models of renal injury and early renal biomarkers that can predict onset of CKD. In the EU FP7 funded project, Predict-IV, we investigated the human renal proximal tubule cells line, RPTEC/TERT1 for their applicability to long term nephrotoxic mechanistic studies. To this end, we used a tiered strategy to optimise dosing regimes for 9 nephrotoxins. Our final testing protocol utilised differentiated RPTEC/TERT1 cells cultured on filter inserts treated with compounds at both the apical and basolateral side, at concentrations not exceeding IC10, for 14 days in a 24 h repeat application. Transepithelial electrical resistance and supernatant lactate were measured over the duration of the experiments and genome wide transcriptomic profiles were assayed at day 1, 3 and 14. The effect of hypoxia was investigated for a subset of compounds. The transcriptomic data were analysed to investigate compound-specific effects, global responses and mechanistically informative signatures. In addition, several potential clinically useful renal injury biomarkers were identified.
authors.affiliation: Division of Physiology, Dept. of Physiology and Medical Physics, Innsbruck Medical University, Fritz-Pregl Strasse 3, 6020 Innsbruck, Austria.
authors.forename: Lydia
authors.initials: L
authors.lastname: Aschauer
authors.affiliation: Division of Physiology, Dept. of Physiology and Medical Physics, Innsbruck Medical University, Fritz-Pregl Strasse 3, 6020 Innsbruck, Austria.
authors.forename: Alice
authors.initials: A
authors.lastname: Limonciel
authors.affiliation: Division of Physiology, Dept. of Physiology and Medical Physics, Innsbruck Medical University, Fritz-Pregl Strasse 3, 6020 Innsbruck, Austria.
authors.forename: Anja
authors.initials: A
authors.lastname: Wilmes
authors.affiliation: Department of Biostatistics, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Stanzel
authors.affiliation: Department of Biostatistics, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
authors.forename: Annette
authors.initials: A
authors.lastname: Kopp-Schneider
authors.affiliation: Non-Clinical Safety, Merck Serono, Frankfurter Strasse 250, 64293 Darmstadt, Germany.
authors.forename: Philip
authors.initials: P
authors.lastname: Hewitt
authors.affiliation: Emergentec Biodevelopment GmbH, Gersthofer Strasse 29-31, 1180 Vienna, Austria.
authors.forename: Arno
authors.initials: A
authors.lastname: Lukas
authors.affiliation: Centre for Radiation, Chemical and Environmental Hazards, Public Health England, Chilton, Didcot, Oxon OX11 0RQ, UK.
authors.forename: Martin O
authors.initials: MO
authors.lastname: Leonard
authors.affiliation: Division of Physiology, Dept. of Physiology and Medical Physics, Innsbruck Medical University, Fritz-Pregl Strasse 3, 6020 Innsbruck, Austria.
authors.forename: Walter
authors.initials: W
authors.lastname: Pfaller
authors.affiliation: Division of Physiology, Dept. of Physiology and Medical Physics, Innsbruck Medical University, Fritz-Pregl Strasse 3, 6020 Innsbruck, Austria. Electronic address: paul.jennings@i-med.ac.at.
authors.forename: Paul
authors.initials: P
authors.lastname: Jennings
mesh_terms: D018929:Cell Culture Techniques; D002460:Cell Line; D017097:Electric Impedance; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007674:Kidney Diseases / Q000139:chemically induced*; D007687:Kidney Tubules, Proximal / Q000166:cytology*; D007773:Lactates / Q000378:metabolism; D004364:Pharmaceutical Preparations; D059467:Transcriptome
chemical_list: D007773:Lactates; D004364:Pharmaceutical Preparations
keywords: Nephrotoxin; Proximal tubule; RPTEC/TERT1; Stress response; Transcriptomics