New three-dimensional preclinical models to understand and treat liver cancers activated for the β-catenin pathway

Background & aims

Constitutive activation of the β-catenin pathway is a determining feature in the pathogenesis of two primary liver cancers, namely HCC and hepatoblastoma (HB). Activating alterations in CTNNB1 gene and, to a lesser extent, inhibiting alterations in APC gene are observed in 30 to 40% of HCC cases and 80 to 90% of HB cases. For both tumours, therapeutic management is far from optimal. Therefore, relevant experimental models are needed to increase our knowledge and test new therapeutic approaches.

Methods

Organoids and tumouroids were established from APC ^Δhep and βcat ^Δex3 mouse models, which are clinically relevant models for β-catenin-activated HCC and mesenchymal HB. We developed a new methodological approach based on a dynamic suspension culture in a rotating bioreactor. Morphological and molecular characteristics and sensitivity to WNTinib, a treatment already successfully tested on human HCC and HB tumouroids, were evaluated by histology, immunohistochemistry, immunofluorescence, and RT-qPCR.

Results

This easy-to-implement methodology allows for the rapid generation of a large number of organoids and tumouroids that are uniform in size and show no signs of cell death in their core. The robustness of the methodology is illustrated by the maintenance of the histological architecture, cell diversity and gene expression in organoids and tumouroids in comparison with the native liver tissues. In addition, the value of the HCC-derived tumouroids for evaluating cancer treatment was assessed based on their responsiveness to the β-catenin antagonist WNTinib.

Conclusions

The organoids and tumouroids that we present here are new reliable in vitro cancer models, recapitulating the main features of β-catenin-driven HCC and mesenchymal HB. They can be integrated into an appropriate platform for drug screening and could enable the development of “à la carte” therapies that are urgently needed for these indications.

Impact and implications

This study addresses the critical need for representative in vitro models to investigate β-catenin-driven liver cancers. The organoids and tumouroids developed here are particularly valuable for researchers seeking robust, reproducible models that accurately reflect the cellular diversity and gene expression profiles of native liver tumours. These findings have practical applications in exploring cancer mechanisms, screening new drugs, optimizing personalized treatment strategies, and reducing reliance on animal models, which ultimately benefits patients.

Highlights

• Easy and rapid generation of mouse liver organoids and tumouroids from β-catenin activated tumours using culture in a bioreactor

• Tumouroids preserve histology, cell diversity, and gene expression of native tissue

• HCC-derived tumouroids respond to β-catenin inhibitor WNTinib

• These reliable 3D models reduce reliance on animal experiments for drug testing