title: An Automated Multiplexed Hepatotoxicity and CYP Induction Assay Using HepaRG Cells in 2D and 3D.
abstract: Drug-induced liver injury (DILI) and drug-drug interactions (DDIs) are concerns when developing safe and efficacious compounds. We have developed an automated multiplex assay to detect hepatotoxicity (i.e., ATP depletion) and metabolism (i.e., cytochrome P450 1A [CYP1A] and cytochrome P450 3A4 [CYP3A4] enzyme activity) in two-dimensional (2D) and three-dimensional (3D) cell cultures. HepaRG cells were cultured in our proprietary micromold plates and produced spheroids. HepaRG cells, in 2D or 3D, expressed liver-specific proteins throughout the culture period, although 3D cultures consistently exhibited higher albumin secretion and CYP1A/CYP3A4 enzyme activity than 2D cultures. Once the spheroid hepatic quality was assessed, 2D and 3D HepaRGs were challenged to a panel of DILI- and CYP-inducing compounds for 7 days. The 3D HepaRG model had a 70% sensitivity to liver toxins at 7 days, while the 2D model had a 60% sensitivity. In both the 2D and 3D HepaRG models, 83% of compounds were predicted to be CYP inducers after 7 days of compound exposure. Combined, our results demonstrate that an automated multiplexed liver spheroid system is a promising cell-based method to evaluate DILI and DDI for early-stage drug discovery.
authors.affiliation: 1 Likarda, LLC, Kansas City, KS, USA.
authors.forename: Lindsey M
authors.initials: LM
authors.lastname: Ott
authors.affiliation: 1 Likarda, LLC, Kansas City, KS, USA.
authors.forename: Karthik
authors.initials: K
authors.lastname: Ramachandran
authors.affiliation: 1 Likarda, LLC, Kansas City, KS, USA.|2 University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Lisa
authors.initials: L
authors.lastname: Stehno-Bittel
mesh_terms: D001681:Biological Assay / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods*; D056486:Chemical and Drug Induced Liver Injury / Q000517:prevention & control; D065693:Cytochrome P-450 Enzyme Inducers / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004347:Drug Interactions / Q000502:physiology; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000187:drug effects*; D018874:Spheroids, Cellular / Q000187:drug effects / Q000378:metabolism
chemical_list: D065693:Cytochrome P-450 Enzyme Inducers; D003577:Cytochrome P-450 Enzyme System
keywords: drug discovery; drug-induced liver injury; drug–drug interaction; scaffold-free; spheroid