Predictive toxicology using systemic biology and liver microfluidic “on chip” approaches: Application to acetaminophen injury

Abstract

Snouber, et al.. Predictive toxicology using systemic biology and liver microfluidic "on chip" approaches: Application to acetaminophen injury. Toxicology and Applied Pharmacology, Elsevier, 2012, 259 (3) ABSTRACTWe -analyzed transcriptomic, proteomic and metabolomic profiles of hepatoma cells cultivated inside a microfluidic biochip with or without acetaminophen (APAP).Without APAP, the results show an adaptive cellular response to the microfluidic environment, leading to the induction of anti-oxidative stress and cytoprotective pathways. In presence of APAP, calcium homeostasis perturbation, lipid peroxidation and cell death are observed. These effects can be attributed to APAP metabolism into its highly reactive metabolite, N-acetyl-p-benzoquinone imine (NAPQI). . That toxicity pathway was confirmed by the detection of GSH-APAP, the large production of 2-hydroxybutyrate and 3-hydroxybutyrate, and methionine, cystine, and histidine consumption in the treated biochips. Those metabolites have been reported as specific biomarkers of hepatotoxicity and glutathione depletion in the literature. In addition, the integration of the metabolomic, transcriptomic and proteomic profiles collected allowed a more complete reconstruction of the APAP injury pathways. To our knowledge, this work is the first example of a global integration of microfluidic biochip data in toxicity assessment. Our results demonstrate the potential of that new approach to predictive toxicology.

Investigation of acetaminophen toxicity in HepG2/C3a microscale cultures using a system biology model of glutathione depletion

“…It also predicted GSH synthesis to detoxify the ROS production. This GSH production appeared consistent with our metabolomic profiling in which we detected in control biochip (when compared to Petri) a production of pyroglutamate and of 2-hydroxybutyrate (Prot et al 2012;Choucha Snouber et al 2012b). Both compounds (not initially inside the culture medium) are reported to be cell stress biomarkers occurring during GSH synthesis (Lu 1999;Ghauri et al 1993;Lord and Bralley 2008).…”

“…The experiments used to calibrate the system biology model and to analyze its behavior during acetaminophen (APAP) exposures resulted from our transcriptomic and proteomic works in microfluidic biochips (Prot et al 2011(Prot et al , 2012. Detailed protocols are given in Supplement File 1.…”

“…When the glutathione is depleted, the NAPQI binds to proteins and leads to necrosis (Reid et al 2005). We previously reported higher acetaminophen toxicity in HepG2/C3a cultivated in liver microfluidic biochip when compared to Petri culture (Prot et al 2011(Prot et al , 2012. In present paper, we have integrated the in vitro information (ROS kinetics, transcriptomic fold changes, viability analysis resulting from Petri and biochips) in the PK-SB model to understand the HepG2/C3a response to acetaminophen exposure in biochips.…”

Judging the value of ‘liver-on-a-chip’ devices for prediction of toxicity

“…Perfusion of the liver chips not only mimics the dynamic flow environment in the liver in vivo and stimulates hepatocyte functions, but it also may improve toxicity prediction of drugs [3,4] and provide control over the exposure to the toxin and removal of waste products. The application of flow to the chip in singlepass mode allows controlling the exposure to the toxin and to test time-dependent variations in the concentration of the toxic compound mimicking a repeated oral dose exposure, which is one of the most challenging issues in safety assessment using in vitro predictive toxicology.…”

The Current Status of Alternatives to Animal Testing and Predictive Toxicology Methods Using Liver Microfluidic Biochips

“…Prot et al 71 have analyzed the integration of the transcriptomics, proteomics and metabonomics profiles of a liver tissue cultivated inside a microfluidic biochip in the absence and presence of acetaminophen (APAP). Without APAP, the results show an adaptive cell response to the microfluidic environment leading to the induction of the anti-oxidative stress and cytoprotective pathways.…”