Comparison of Liver Cell Models Using the Basel Phenotyping Cocktail

Benjamin Berger,Stephan Krähenbühl,Adrian Roth,Franziska Boess,Massimiliano Donzelli,Swarna Maseneni,Manuel Haschke

doi:10.3389/fphar.2016.00443

Benjamin Berger, Stephan Krähenbühl + Show 5 more

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https://doi.org/10.3389/fphar.2016.00443

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Abstract

Currently used hepatocyte cell systems for in vitro assessment of drug metabolism include hepatoma cell lines and primary human hepatocyte (PHH) cultures. We investigated the suitability of the validated in vivo Basel phenotyping cocktail (caffeine [CYP1A2], efavirenz [CYP2B6], losartan [CYP2C9], omeprazole [CYP2C19], metoprolol [CYP2D6], midazolam [CYP3A4]) in vitro and characterized four hepatocyte cell systems (HepG2 cells, HepaRG cells, and primary cryopreserved human hepatocytes in 2-dimensional [2D] culture or in 3D-spheroid co-culture) regarding basal metabolism and CYP inducibility. Under non-induced conditions, all CYP activities could be determined in 3D-PHH, CYP2B6, CYP2C19, CYP2D6, and CYP3A4 in 2D-PHH and HepaRG, and CYP2C19 and CYP3A4 in HepG2 cells. The highest non-induced CYP activities were observed in 3D-PHH and HepaRG cells. mRNA expression was at least four-fold higher for all CYPs in 3D-PHH compared to the other cell systems. After treatment with 20 μM rifampicin, mRNA increased 3- to 50-fold for all CYPs except CYP1A2 and 2D6 for HepaRG and 3D-PHH, 4-fold (CYP2B6) and 17-fold (CYP3A4) for 2D-PHH and four-fold (CYP3A4) for HepG2. In 3D-PHH at least a two-fold increase in CYP activity was observed for all inducible CYP isoforms while CYP1A2 and CYP2C9 activity did not increase in 2D-PHH and HepaRG. CYP inducibility assessed in vivo using the same phenotyping probes was also best reflected by the 3D-PHH model. Our studies show that 3D-PHH and (with some limitations) HepaRG are suitable cell systems for assessing drug metabolism and CYP induction in vitro. HepG2 cells are less suited to assess CYP induction of the 2C and 3A family. The Basel phenotyping cocktail is suitable for the assessment of CYP activity and induction also in vitro.

Highlights

Cytochrome P450 enzymes (CYPs) are involved in the oxidative metabolism of the majority of the commonly used low molecular weight drugs, thereby influencing the pharmacokinetics of these drugs and having an important role in drug-drug interactions (Wilkinson, 2005)
In comparison to 3D-cultured primary cryopreserved human hepatocytes (PHH), HepaRG cells showed a 4 times lower mRNA expression of CYP3A4 and 2C19, whereas the mRNA expression of the other CYPs investigated was more than 10 times lower
This study confirms that basal mRNA expression and CYP activity as well as CYP inducibility show large differences between in vitro used hepatocyte cell models and, depending on the hepatocyte model used, that the in vivo characterized Basel phenotyping cocktail is suitable for CYP induction experiments in vitro

Summary

Introduction

Cytochrome P450 enzymes (CYPs) are involved in the oxidative metabolism of the majority of the commonly used low molecular weight drugs, thereby influencing the pharmacokinetics of these drugs and having an important role in drug-drug interactions (Wilkinson, 2005). Several cocktails have been characterized in clinical studies and are used in individual patients, including the Karolinska cocktail (Christensen et al, 2003), the Cooperstown 5+1 cocktail (Chainuvati et al, 2003), the Geneva cocktail (Bosilkovska et al, 2014) and the Basel Cocktail (Donzelli et al, 2014). These cocktails have been shown to be valuable tools to assess the capacity of drugs to induce or inhibit CYPs (Bosilkovska et al, 2014; Derungs et al, 2016). Drug cocktails have been used to perform in vitro studies, in particular when assessing the CYP inhibition and induction potential of chemical compounds (Youdim et al, 2007; Mori et al, 2009; Spaggiari et al, 2014)

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