Human-correlated genetic HCC models identify combination therapy for precision medicine

Abstract

Hepatocellular carcinoma (HCC), the most common form of primary liver cancer, is a leading cause of cancer related mortality worldwide. HCC occurs typically from a background of chronic liver disease, caused by a spectrum of predisposing conditions. Tumour development is driven by the expansion of clones that accumulated progressive driver mutations, with hepatocytes the most likely cell of origin. However, the landscape of driver mutations in HCC is independent of the underlying aetiologies. Despite an increasing range of systemic treatment options for advanced HCC outcomes remain heterogeneous and typically poor. Emerging data suggest that drug efficacies depend on disease aetiology and genetic alterations. Exploring subtypes in preclinical models with human relevance will therefore be essential to advance precision medicine in HCC. We generated over twenty-five new genetically-driven in vivo and in vitro HCC models. Our models represent multiple features of human HCC, including clonal origin, histopathological appearance, and metastasis to distant organs. We integrated transcriptomic data from the mouse models with human HCC data and identified four common human-mouse subtype clusters. The subtype clusters had distinct transcriptomic characteristics that aligned with histopathology. In a proof-of-principle analysis, we verified response to standard of care treatment and used a linked in vitro-in vivo pipeline to identify a promising therapeutic candidate, cladribine, that has not been linked to HCC treatment before. Cladribine acts in a highly effective subtype-specific manner in combination with standard of care therapy.

The eIF4A2 negative regulator of mRNA translation promotes extracellular matrix deposition to accelerate hepatocellular carcinoma initiation

“…To model initiation and progression of HCC in vivo, we used mice carrying floxed alleles of constitutively active β-catenin (Ctnnb1 ex3flox/WT ) and additional copies of the MYC oncogene (Rosa26 LSL-MYC/LSL-MYC ). Expression of these oncogenes was initiated by injecting mice with AAV8-TBG-Cre at a titer that achieves recombination in a small proportion (≈5%) of hepatocytes; the tumour cell-of-origin for HCC (18,19) (Fig. 1A).…”

Multi-scale in vivo imaging of tumour development using a germline conditional triple-reporter system

“…A critical advantage of developing an inducible multiscale imaging reporter system is to track the presence, expansion, and movement of cell populations over time. We therefore tested the utility of the R26 LSL − NRL reporter allele in the context of tumour formation in a conditional mouse model of hepatocellular carcinoma combining two gain-of-function Cre-inducible alleles Ctnnb1 ex3/wt and R26 LSL − MYC/wt (BM) [33,39]. We induced liver tumours by intravenous injection of AAV8-TBG-Cre into NRL-BM and BM (littermate control) mice (Fig.…”

“…NRL-CV: R26 wt/LSL-NRL + CMV-Cre NRL-KP: R26 LSL-NRL/wt + Kras LSL-G12D/wt + Trp53 / KP: Kras LSL-G12D/wt + Trp53 / NRL-BM: Ctnnb1 ex3/wt + R26 LSL-MYC/LSL-NRL BM: Ctnnb1 ex3/wt + R26 LSL-MYC/wt Tumour induction Liver tumours were induced in 8-10 week old, male NRL-BM or BM littermate control mice using AAV8.TBG.PI.Cre.rBG (AAV8-TBG-Cre; Addgene#107787-AAV8) viral vector (Adeno-associated virus 8 with payload under Thyroxin Binding Globulin promoter with hepatocyte tropism) [33]. Control NRL-BM animals were infected with AAV8.TBG.PI.Null.bGH virus (AAV8-TBG-Null; Addgene#105536-AAV8) serving as a genetic control.…”

Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer

“…Publisher's note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. 32 . Mice were randomly assigned to experimental groups based on their genotypes, and, with the exception of MRI quantification, the analyses were not performed blindly.…”