The development of hepatocellular carcinoma models in precision-cut tissue slices for therapeutic screening and precision medicine

Abstract

Liver cancer is the third most common cause of cancer-related death worldwide, and hepatocellular carcinoma (HCC) accounts for approximately 90% of cases. Recent therapeutic advances extend overall survival by a few months, but only for a minority of patients. Realistic HCC models are necessary in order to provide valuable insight into disease pathogenesis and drug discovery. Here I describe the development of two HCC models in precision-cut liver slices (PCLS): a murine precision-cut tumour slice (PCTS) model and a spheroid-engrafted human PCLS model. An orthotopic mouse model of HCC was generated using the Hep-53.4 cell line, and PCTS were subsequently generated from the tumours. Histological characterisation determined that Hep-53.4 PCTS retain the histological characteristics of in vivo tumours, and mimic the in vivo therapeutic responses to sorafenib and lenvatinib. Intervention with anti-PD1 immunotherapy significantly increased CD3 T cell numbers and apoptosis – a response that was lost upon lipid-loading of the PCTS. After initial optimisation using murine PCLS and Hep-53.4 cells, a human HCC model was developed by engrafting spheroids generated from HuH7 cells that express a secreted luciferase onto human PCLS. Complete invasion of the spheroids into the PCLS was confirmed via multiphoton imaging, and measurement of luciferase secreted following treatment with tyrosine kinase inhibitors indicated a significant and dose-dependent reduction in cancer growth, whilst the PCLS remained viable. A patient-derived HCC cell line library was generated to tailor the spheroidPCLS model for precision medicine. Hyperion imaging mass cytometry revealed that both the HuH7 and patient-derived spheroid-PCLS systems stimulated a response from immune components in the surrounding PCLS, alongside potential matrix formation within the HCC spheroid. Thus, the ex vivo models developed potentially present as useful tools for discovery biology and precision medicine, where therapies can be tested on patient-derived HCC cells in the context of the tumour microenvironment.