Interferon- Overexpression in Adipose Tissue-Derived Stem Cells Induces HepG2 and Macrophage Cell Death in Liver Tumor Organoids via Induction of TNF-Related Apoptosis-Inducing Ligand Expression

Liver tumor organoids derived from liver tumor tissues and pluripotent stem cells are used for liver tumor research but have several challenges in primary cell isolation and stem cell differentiation. Here, we investigated the potential of HepG2-based liver tumor organoids for screening anticancer drugs by evaluating their responsiveness to IFN-beta produced by mesenchymal stem cells (MSCs). Liver tumor organoids were prepared in three days on Matrigel using HepG2, primary liver sinusoidal epithelial cells (LSECs), LX-2 human hepatic stellate cells, and THP-1-derived macrophages at a ratio of 4:4:1:1, with 105 total cells. Hepatocyte-related and M2 macrophage-associated genes increased in liver tumor organoids. IFN-beta treatment decreased the viability of liver tumor organoids and increased M1 macrophage marker expression (i.e., TNF-alpha and iNOS) and TRAIL. TRAIL expression was increased in all four cell types exposed to IFN-beta, but cell death was only observed in HepG2 cells and macrophages. Further, MSCs overexpressing IFN-beta (ASC-IFN-beta) also expressed TRAIL, contributing to the reduced viability of liver tumor organoids. In summary, IFN-beta or ASC-IFN-beta can induce TRAIL-dependent HepG2 and macrophage cell death in HepG2-based liver tumor organoids, highlighting these liver tumor organoids as suitable for anticancer drug screening and mechanistic studies.