Molecular characterisation of hepatocellular carcinoma in patients with non-alcoholic steatohepatitis

Roser Pinyol, Sara Torrecilla, Huan Wang, Carla Montironi, Marta Piqué-Gili, Miguel Torres-Martin, Leow Wei-Qiang, Catherine E. Willoughby, Pierluigi Ramadori, Carmen Andreu-Oller, Patricia Taik, Youngmin A. Lee, Agrin Moeini, Judit Peix, Suzanne Faure-Dupuy, Tobias Riedl, Svenja Schuehle, Claudia P. Oliveira, Venancio A. Alves, Paolo BoffettaAnja Lachenmayer, Stephanie Roessler, Beatriz Minguez, Peter Schirmacher, Jean François Dufour, Swan N. Thung, Helen L. Reeves, Flair J. Carrilho, Charissa Chang, Andrew V. Uzilov, Mathias Heikenwalder, Arun Sanyal, Scott L. Friedman, Daniela Sia, Josep M. Llovet

Research output: Contribution to journalArticlepeer-review

70 Scopus citations


Background and Aims: Non-alcoholic steatohepatitis (NASH)-related hepatocellular carcinoma (HCC) is increasing globally, but its molecular features are not well defined. We aimed to identify unique molecular traits characterising NASH-HCC compared to other HCC aetiologies. Methods: We collected 80 NASH-HCC and 125 NASH samples from 5 institutions. Expression array (n = 53 NASH-HCC; n = 74 NASH) and whole exome sequencing (n = 52 NASH-HCC) data were compared to HCCs of other aetiologies (n = 184). Three NASH-HCC mouse models were analysed by RNA-seq/expression-array (n = 20). Activin A receptor type 2A (ACVR2A) was silenced in HCC cells and proliferation assessed by colorimetric and colony formation assays. Results: Mutational profiling of NASH-HCC tumours revealed TERT promoter (56%), CTNNB1 (28%), TP53 (18%) and ACVR2A (10%) as the most frequently mutated genes. ACVR2A mutation rates were higher in NASH-HCC than in other HCC aetiologies (10% vs. 3%, p <0.05). In vitro, ACVR2A silencing prompted a significant increase in cell proliferation in HCC cells. We identified a novel mutational signature (MutSig-NASH-HCC) significantly associated with NASH-HCC (16% vs. 2% in viral/alcohol-HCC, p = 0.03). Tumour mutational burden was higher in non-cirrhotic than in cirrhotic NASH-HCCs (1.45 vs. 0.94 mutations/megabase; p <0.0017). Compared to other aetiologies of HCC, NASH-HCCs were enriched in bile and fatty acid signalling, oxidative stress and inflammation, and presented a higher fraction of Wnt/TGF-β proliferation subclass tumours (42% vs. 26%, p = 0.01) and a lower prevalence of the CTNNB1 subclass. Compared to other aetiologies, NASH-HCC showed a significantly higher prevalence of an immunosuppressive cancer field. In 3 murine models of NASH-HCC, key features of human NASH-HCC were preserved. Conclusions: NASH-HCCs display unique molecular features including higher rates of ACVR2A mutations and the presence of a newly identified mutational signature. Lay summary: The prevalence of hepatocellular carcinoma (HCC) associated with non-alcoholic steatohepatitis (NASH) is increasing globally, but its molecular traits are not well characterised. In this study, we uncovered higher rates of ACVR2A mutations (10%) – a potential tumour suppressor – and the presence of a novel mutational signature that characterises NASH-related HCC.

Original languageEnglish
Pages (from-to)865-878
Number of pages14
JournalJournal of Hepatology
Issue number4
StatePublished - Oct 2021


  • animal model
  • liver cancer
  • metabolic syndrome
  • molecular class
  • mutational signature
  • obesity


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