3D cellular models are now becoming commonplace within the pharmaceutical industry, especially for more complex issues such as the prediction of drug-induced toxicity. In 2017, Genentech authored a publication on the prediction of human DILI (drug-induced liver injury) using primary human hepatocyte 3D liver microtissues. The approach compared 2D vs 3D models and demonstrated the value of the 3D model in terms of improved predictive capability over the 2D approach. The study focused on a single ATP endpoint.
Research at Cyprotex has extended this approach significantly using a comprehensive panel of endpoints including mitochondrial membrane potential, reactive oxygen species formation and glutathione content in addition to cellular ATP. Two 3D cell-based models were assessed including a HepaRG monoculture liver model and a co-cultured human hepatocyte and human non-parenchymal cell model. The microtissues were formed using scaffold-free ultra low attachment round bottomed plates. Cytochrome P450 activity was evaluated in the 3D models using a range of different selective probe substrates.
By utilising multi-parametric high content screening (HCS) in addition to ATP, a significantly improved sensitivity was observed over ATP content alone while sensitivity remained similar. Furthermore, the HepaRG 3D microtissues appeared to provide an increased sensitivity to predict DILI-positive compounds compared to the co-culture 3D human hepatocyte model. This may be related to the increased level of cytochrome P450 activity in the HepaRG 3D microtissues compared to the 3D human hepatocyte model.
Our research was presented at SOT in San Antonio from March 11-15, 2018.