S5: Establishment of transcriptomic database based on multi-organ interaction and screening the predictive biomarkers
Establishment of transcriptomic database based on multi-organ interaction and screening the predictive biomarkers for drug-induced nephrotoxicity using co-culture system
To mimic the systemic effect like organ-organ interaction, we have developed the co-culture platforms including liver, kidney and endothelial cell culture. Using in vitro kidney cells, the effects of metabolic activation under liver could not be reflected in the only kidney cell culture system. To reflect the metabolic activation under liver and interaction with endothelial cells during nephrotoxicity, we have screened nephrotoxic chemicals under co-culture platforms with hepatic metabolism. We established the CYP overexpressed HepG2 cell lines (HepG2/CYPs) which stably co-express the major CYPs enzymes, CYP3A4, CYP1A2, CYP2B6, and CYP2E1. Using HepG2/CYPs, HK-2, and HUVEC, we optimized the co-culture assay system for evaluating the nephrotoxicity. Using this co-culture platforms, we performed the transcriptomic approaches and established local database for potentially toxic chemicals in the kidney. The in vitro potential nephrotoxic biomarkers are identified by genomic analysis of HepG2/CYP, HK-2, and HUVEC cells after exposure of 10 different nephrotoxic chemicals. This data may contribute in elucidation of the molecular events during in vitro kidney cells after activation of hepatic metabolism and to discover the predictive biomarkers for screening the drug-induced nephrotoxicity.