Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress.

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 characterized nephrotoxin Cyclosporine A (CsA) at therapeutic and supra therapeutic concentrations for 14 days. 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 15 µM 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 5 µM 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.

WILMES Anja;  LIMONCIEL Alice;  ASCHAUER Lydia;  MOENKS Konrad;  BIELOW Chris;  LEONARD Matrin;  HAMON Jeremy;  CARPI Donatella;  RUZEK Silke;  HANDLER Andreas;  SCHMAL Olga;  HERRGEN Karin;  BELLWON Patricia;  BUREK Christof;  TRUISI Germaine;  HEWITT Philip;  DI CONSIGLIO Emma;  TESTAI Emanuela;  BLAAUBOER Bas J.;  GUILLOU Claude;  HUBER Christian;  LUKAS Arno;  PFALLER Walter;  MUELLER Stefan;  BOIS Frederic;  DEKANT Wolfgang;  JENNINGS Paul; 

2013-03-06

ELSEVIER SCIENCE BV

JRC77136

1874-3919,   

http://www.sciencedirect.com/science/article/pii/S1874391912007774,    https://publications.jrc.ec.europa.eu/repository/handle/JRC77136,   

10.1016/j.jprot.2012.11.022,