Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG cell-based biosensors to monitor hepatocyte differentiation, drug metabolism and toxicity

Manuel Vlach (1) , Nicolas Quesnot (1) , Hélène Dubois-Pot-Schneider (2) , Catherine Ribault (1) , Yann Verres (1) , Kilian Petitjean (1) , Claudine Rauch (1) , Fabrice Morel (1) , Marie-Anne Robin (1) , Anne Corlu (1) , Pascal Loyer (1)

1 NuMeCan - Nutrition, Métabolismes et Cancer
2 CRAN - Centre de Recherche en Automatique de Nancy

Human hepatoma HepaRG cells express most drug metabolizing enzymes and constitute a pertinent in vitro alternative cell system to primary cultures of human hepatocytes in order to determine drug metabolism and evaluate the toxicity of xenobiotics. In this work, we established novel transgenic HepaRG cells transduced with lentiviruses encoding the reporter green fluorescent protein (GFP) transcriptionally regulated by promoter sequences of cytochromes P450 (CYP) 1A1/2, 2B6 and 3A4 genes. Here, we demonstrated that GFP-biosensor transgenes shared similar expression patterns with the corresponding endogenous CYP genes during proliferation and differentiation in HepaRG cells. Interestingly, differentiated hepatocyte-like HepaRG cells expressed GFP at higher levels than cholangiocyte-like cells. Despite weaker inductions of GFP expression compared to the strong increases in mRNA levels of endogenous genes, we also demonstrated that the biosensor transgenes were induced by prototypical drug inducers benzo(a)pyrene and phenobarbital. In addition, we used the differentiated biosensor HepaRG cells to evidence that pesticide mancozeb triggered selective cytotoxicity of hepatocyte-like cells. Our data demonstrate that these new biosensor HepaRG cells have potential applications in the field of chemicals safety evaluation and the assessment of drug hepatotoxicity.

Mots clés: cytochromes P450; hepatotoxicity; drug screening; HepaRG cells; cell-based biosensors

Domaines: Sciences du Vivant [q-bio]

Manuel Vlach, Nicolas Quesnot, Hélène Dubois-Pot-Schneider, Catherine Ribault, Yann Verres, et al.. Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG cell-based biosensors to monitor hepatocyte differentiation, drug metabolism and toxicity. Sensors, 2019, 19 (10), pp.2245. ⟨10.3390/s19102245⟩. ⟨hal-02152870⟩