title: Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress.
abstract: High content omic techniques in combination with stable human in vitro cell culture systems have the potential to improve on current pre-clinical safety regimes by providing detailed mechanistic information of altered cellular processes. Here we investigated the added benefit of integrating transcriptomics, proteomics and metabolomics together with pharmacokinetics for drug testing regimes. Cultured human renal epithelial cells (RPTEC/TERT1) were exposed to the nephrotoxin Cyclosporine A (CsA) at therapeutic and supratherapeutic concentrations for 14days. CsA was quantified in supernatants and cellular lysates by LC-MS/MS for kinetic modeling. There was a rapid cellular uptake and accumulation of CsA, with a non-linear relationship between intracellular and applied concentrations. CsA at 15μM induced mitochondrial disturbances and activation of the Nrf2-oxidative-damage and the unfolded protein-response pathways. All three omic streams provided complementary information, especially pertaining to Nrf2 and ATF4 activation. No stress induction was detected with 5μM CsA; however, both concentrations resulted in a maximal secretion of cyclophilin B. The study demonstrates for the first time that CsA-induced stress is not directly linked to its primary pharmacology. In addition we demonstrate the power of integrated omics for the elucidation of signaling cascades brought about by compound induced cell stress.
authors.affiliation: Division of Physiology, Department of Physiology and Medical Physics, Innsbruck Medical University, Innsbruck 6020, Austria.
authors.forename: Anja
authors.initials: A
authors.lastname: Wilmes
authors.forename: Alice
authors.initials: A
authors.lastname: Limonciel
authors.forename: Lydia
authors.initials: L
authors.lastname: Aschauer
authors.forename: Konrad
authors.initials: K
authors.lastname: Moenks
authors.forename: Chris
authors.initials: C
authors.lastname: Bielow
authors.forename: Martin O
authors.initials: MO
authors.lastname: Leonard
authors.forename: Jeremy
authors.initials: J
authors.lastname: Hamon
authors.forename: Donatella
authors.initials: D
authors.lastname: Carpi
authors.forename: Silke
authors.initials: S
authors.lastname: Ruzek
authors.forename: Andreas
authors.initials: A
authors.lastname: Handler
authors.forename: Olga
authors.initials: O
authors.lastname: Schmal
authors.forename: Karin
authors.initials: K
authors.lastname: Herrgen
authors.forename: Patricia
authors.initials: P
authors.lastname: Bellwon
authors.forename: Christof
authors.initials: C
authors.lastname: Burek
authors.forename: Germaine L
authors.initials: GL
authors.lastname: Truisi
authors.forename: Philip
authors.initials: P
authors.lastname: Hewitt
authors.forename: Emma
authors.initials: E
authors.lastname: Di Consiglio
authors.forename: Emanuela
authors.initials: E
authors.lastname: Testai
authors.forename: Bas J
authors.initials: BJ
authors.lastname: Blaauboer
authors.forename: Claude
authors.initials: C
authors.lastname: Guillou
authors.forename: Christian G
authors.initials: CG
authors.lastname: Huber
authors.forename: Arno
authors.initials: A
authors.lastname: Lukas
authors.forename: Walter
authors.initials: W
authors.lastname: Pfaller
authors.forename: Stefan O
authors.initials: SO
authors.lastname: Mueller
authors.forename: Frederic Y
authors.initials: FY
authors.lastname: Bois
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Dekant
authors.forename: Paul
authors.initials: P
authors.lastname: Jennings
mesh_terms: D021983:Cyclophilins / Q000378:metabolism; D016572:Cyclosporine / Q000493:pharmacokinetics*; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000166:cytology; D055432:Metabolomics; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D040901:Proteomics; D015398:Signal Transduction / Q000187:drug effects; D053719:Tandem Mass Spectrometry; D014116:Toxicology / Q000379:methods
chemical_list: D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; C081786:cyclophilin B; D016572:Cyclosporine; D021983:Cyclophilins