Human In-vitro Inflammatory Liver Model Recapitulates Immune-Associated Drug Efects with High Predictivity

Immune-mediated drug-induced liver injury is often undermined or not well-studied due to lack of appropriate in vitro models comprised of immune cells and suitable endpoints. Current advances in co-cultures deploying primary human Kupffer cells (PHKCs) or alternative cells such as THP-1 macrophages are limited by their inability of recapitulating drug-induced cytokine responses which are important immune-mediated drug effects. Since human iPSC-derived Kupffer cells (iKCs, PMID: 30497023) functionally resemble PHKCs, we investigated the use of iKCs to mimic drug-specific cytokine responses in vitro. iKCs were co-cultured with hiPSC-derived hepatocytes, which were then subjected to treatment of drugs with known immune-mediated effects. Out of the cytokine measurements including interleukin-6 and tumor necrosis factor α obtained from 19 compounds, 17 showed good in vitro-in vivo correlation (89.5%). Furthermore, co-cultures using THP-1 macrophages instead of iKCs could not recapitulate IL-6 responses upon treatment with paradigm compounds. RNA-Seq was used to explore the interaction between cytokines and drug-induced hepatotoxicity in greater detail for one pair of drugs - trovafloxacin and its non-hepatotoxic analogue, levofloxacin. Our study demonstrates for the first time, the use of iKCs in a physiologically relevant model that captures cytokine responses upon treatment with a wide range of immune-mediated drugs and hence can be very useful in warranting further investigation of new drugs that elicit drug-induced cytokine imbalance. Human pluripotent stem cells were differentiated into hepatocytes (iHeps) and Kupffer cells (iKCs). On the first day, iKCs were seeded, followed by iHeps 24h later. On the third day, the co-culture was subjected to drug treatment for 48h. RNAseq was performed on 12 sample types with 3 biological replicates each (except mTVX+ which only has 2 replicates). The sample types are named as . E.g. mTVX+ indicates an iHep monoculture treated with trovafloxacin and LPS.