title: Integrated proteomic and transcriptomic investigation of the acetaminophen toxicity in liver microfluidic biochip.
abstract: Microfluidic bioartificial organs allow the reproduction of in vivo-like properties such as cell culture in a 3D dynamical micro environment. In this work, we established a method and a protocol for performing a toxicogenomic analysis of HepG2/C3A cultivated in a microfluidic biochip. Transcriptomic and proteomic analyses have shown the induction of the NRF2 pathway and the related drug metabolism pathways when the HepG2/C3A cells were cultivated in the biochip. The induction of those pathways in the biochip enhanced the metabolism of the N-acetyl-p-aminophenol drug (acetaminophen-APAP) when compared to Petri cultures. Thus, we observed 50% growth inhibition of cell proliferation at 1 mM in the biochip, which appeared similar to human plasmatic toxic concentrations reported at 2 mM. The metabolic signature of APAP toxicity in the biochip showed similar biomarkers as those reported in vivo, such as the calcium homeostasis, lipid metabolism and reorganization of the cytoskeleton, at the transcriptome and proteome levels (which was not the case in Petri dishes). These results demonstrate a specific molecular signature for acetaminophen at transcriptomic and proteomic levels closed to situations found in vivo. Interestingly, a common component of the signature of the APAP molecule was identified in Petri and biochip cultures via the perturbations of the DNA replication and cell cycle. These findings provide an important insight into the use of microfluidic biochips as new tools in biomarker research in pharmaceutical drug studies and predictive toxicity investigations.
authors.affiliation: CNRS UMR 6600, Laboratoire de Biomécanique et Bioingénierie, Université de Technologie de Compiègne, Compiègne, France.
authors.forename: Jean Matthieu
authors.initials: JM
authors.lastname: Prot
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Briffaut
authors.forename: Franck
authors.initials: F
authors.lastname: Letourneur
authors.forename: Philippe
authors.initials: P
authors.lastname: Chafey
authors.forename: Franck
authors.initials: F
authors.lastname: Merlier
authors.forename: Yves
authors.initials: Y
authors.lastname: Grandvalet
authors.forename: Cécile
authors.initials: C
authors.lastname: Legallais
authors.forename: Eric
authors.initials: E
authors.lastname: Leclerc
mesh_terms: D000082:Acetaminophen / Q000378:metabolism / Q000494:pharmacology*; D018712:Analgesics, Non-Narcotic / Q000378:metabolism / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods; D056945:Hep G2 Cells; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D025341:Principal Component Analysis; D040901:Proteomics / Q000379:methods*; D016196:S Phase / Q000187:drug effects; D059467:Transcriptome; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods
chemical_list: D018712:Analgesics, Non-Narcotic; D000082:Acetaminophen