Derivation of Genetically-Defined Murine Hepatoblastoma Cell Lines with Angiogenic Potential

Most hepatoblastomas (HB) are associated with aberrant expression of ß-catenin (B), YAP (Y) and/or NRF2 (N) transcription factors and can be modeled in mice by over-expressing pairwise of triple combination of these. Virtually no human or murine HB cell lines exist that bear these mutations. We describe here an efficient way to generate cell lines from primary BN and YN tumors. Moreover, one of the BN lines shows a remarkable ability to trans-differentiate into endothelial cells under hypoxic conditions that may facilitate angiogenesis. These cell lines along with previousl-derived BN and BYN lines showed similar sensitivities to drugs commonly used to treat HB. Because the approach for cell line derivation we describe is quite general, it should allow for the generation of additional lines driven by less common factors. A collection of such permanent and well-characterized cell lines will facilitate studies that are difficult or impractical to perform in vivo Overall design: Total RNAs were isolated as previously described using RNeasy columns (Qiagen). Only samples with RIN values >8.0 were processed further. Library preparation and sequencing were performed at the Pittsburgh Liver Research Center's Core Sequencing laboratory (https://livercenter.pitt.edu/gsbc/#Sequencing) using an AVITI/Illumina instrument. Analyses were performed using nf-core/rnaseq version 3.12.0 as previously described. Genes with low expression (average counts among all groups < 1) were filtered out. Normalized counts were generated by DESeq2 and then used for statistical quantification and heat map generation.