We have integrated in vitro and in silico information to investigate acetaminophen (APAP) and its metabolite N-acetyl-p-benzoquinone imine (NAPQI) toxicity in liver biochip. In previous works, we observed higher cytotoxicity of HepG2/C3a cultivated in biochips when exposed to 1 mM of APAP for 72 h as compared to Petri cultures. We complete our investigation with the present in silico approach to extend the mechanistic interpretation of the intracellular kinetics of the toxicity process. For that purpose, we propose a mathematical model based on the coupling of a drug pharmacokinetic model (PK) with a systemic biology model (SB) describing the reactive oxygen species (ROS) production by NAPQI and the subsequent glutathione (GSH) depletion. The SB model was parameterized using (i) transcriptomic data, (ii) qualitative results of time lapses ROS fluorescent curves for both control and 1-mM APAP-treated experiments, and (iii) additional GSH literature data. The PK model was parameterized (i) using the in vitro kinetic data (at 160 μM, 1 mM, 10 mM), (ii) using the parameters resulting from a physiologically based pharmacokinetic (PBPK) literature model for APAP, and (iii) by literature data describing NAPQI formation. The PK-SB model predicted a ROS increase and GSH depletion due to the NAPQI formation. The transition from a detoxification phase and NAPQI and ROS accumulation was predicted for a NAPQI concentration ranging between 0.025 and 0.25 μM in the cytosol. In parallel, we performed a dose response analysis in biochips that shows a reduction of the final hepatic cell number appeared in agreement with the time and doses associated with the switch of the NAPQI detoxification/accumulation. As a result, we were able to correlate in vitro extracellular APAP exposures with an intracellular in silico ROS accumulation using an integration of a coupled mathematical and experimental liver on chip approach.
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Aryl hydrocarbon receptor
Cytochrome P450 gene
Glutamate-cysteine ligase catalityc subunit gene
Glutamate-cysteine ligase regulatory subunit gene
Glutathione peroxidase gene
Glutathione synthetase gene
Glutathione S transferase gene
Kelch-like ECH-associated protein 1
Multidrug resistance-associated protein gene
Nuclear factor (erythroid-derived 2)-like 2 gene
Protein code by NFE2L2 gene
Physiologically based pharmacokinetic
Reactive oxygen species
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This work was carried out in the framework of the Labex MS2T, which was funded by the French Government, through the program “Investments for the future” managed by the National Agency for Research (ANR-11-IDEX-0004-02). The work was supported by the “fondation UTC pour la recherche” via the chaire in predictive toxicology and the project “Puces à cellules 3D”
Conflict of interest
We declare no conflict of interest.
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Leclerc, E., Hamon, J., Claude, I. et al. Investigation of acetaminophen toxicity in HepG2/C3a microscale cultures using a system biology model of glutathione depletion. Cell Biol Toxicol 31, 173–185 (2015). https://doi.org/10.1007/s10565-015-9302-0