AUTHOR=Elcombe Matthew , Mathur Ayesha , Wallace Stephanie , Wright Jayne , Plummer Simon TITLE=Artificial intelligence driven image analysis identifies phenobarbital induced hepatocyte hypertrophy in liver microtissues across species JOURNAL=Frontiers in Drug Discovery VOLUME=4 YEAR=2024 URL=https://www.frontiersin.org/journals/drug-discovery/articles/10.3389/fddsv.2024.1472522 DOI=10.3389/fddsv.2024.1472522 ISSN=2674-0338 ABSTRACT=

In vivo treatment of rats with phenobarbital (PB) induces liver enzyme induction associated with hepatocyte hypertrophy. In the present study we used a novel microTMA technology coupled with artificial intelligence (AI) driven image analysis and proteomics analysis to test the hypothesis that PB treatment of rat and human liver microtissues could recapitulate hepatocyte hypertrophy in vitro. Human and rat liver microtissues were treated with PB over a range of concentrations (500 uM - 2000 uM). Fixed liver microtissues were embedded in paraffin in a microTMA mold, sectioned and stained on parallel microTMA sections with H&E and cell type specific markers, respectively. An AI algorithm was trained to identify and measure changes in hepatocyte cytoplasmic area on images of H&E stained microtissue sections. Image analysis with this algorithm showed that treatment of human and rat liver microtissues with PB (500 uM) for 96 h caused significant increases (p ≤ 0.08- p < 0.01) in hepatocyte cytoplasmic area, a hallmark of hypertrophy. Proteomics analysis of control and PB treated liver microtissue samples confirmed this treatment also caused phase1 and phase 2 enzyme induction in both human and rat samples. In conclusion AI driven image analysis of H&E stained liver microtissue FFPE sections shows that this model can recapitulate a PB-induced hypertrophy response.