High frequency of telomerase reverse transcriptase (tert) promoter somatic mutations in hepatocellular carcinoma and pre-neoplastic lesions
Jean-Charles Nault1, Maxime Mallet1, Camilla Pilati1, Julien Calderaro2, Paulette Bioulac-Sage3, Christophe Laurent4, Alexis Laurent5, Daniel Cherqui6, Charles Balabaud7, Jessica Zucman-Rossi1;
1Inserm U674, Paris, France; 2InsermU674, Pathology department, Henri mondior, Creteil, France; 3Pathology department CHU, Bordeaux, France; 4Surgery department CHU, Bordeaux, France; 5Surgery department CHU Henri Mondor, Creteil, France; 6Surgery department Hepatobiliary center Paul Brousse, Villejuif, France; 7INSERM U1053, Bordeaux, France
Introduction: TERT is involved in the maintenance of telomere length and its reactivation is required in liver tumorigenesis. Recently, somatic mutations of the TERT promoter leading to TERT activation were identified in melanoma. We aim to search for TERT promoter mutations in hepatocellular tumors and study its role in cirrhotic (macronodules without and with dysplasia-hepatocellular carcinoma (HCC) sequence) and non-cirrhotic (hepatocellular adenoma (HCA)-HCC sequence) multistep car-cinogenesis. Methods: We sequenced the TERT promoter in 305 HCC, 20 cirrhotic macronodules, 60 classical HCA and 16 HCA with signs of malignant transformation. These tumors were characterized at clinical and histological level and for the classical genes involved in liver tumorogenesis. We assessed TERT expression using quantitative RT-PCR in 309 hepatocellular tumors and non-tumor liver tissues. Results: we identified somatic mutations of the TERT promoter in 179 (59%) among 305 human HCC. These mutations were localized at the two hot spot described in melanoma (-124G>A and -146G>A from the ATG site). TERT promoter mutations were significantly associated with CTNNB1 mutations (P<0.0001) and were more frequent in small HCC (< 5cm), with low serum AFP level and non-related to chronic HBV infection. TERT expression was significantly higher in HCC with TERT promoter mutations compared to normal liver and cirrhosis (P=0.0007). Additionally, we identified TERT promoter mutations in 25% (5/20) of cirrhotic macronodules with or without dysplasia. In contrast, we didn't found any mutations among 15 genes (CTNNB1, TP53...) classically mutated in HCC and screened in these pre-neoplastic lesions. Interestingly, cirrhotic macronodules mutated for the TERT promoter exhibited an increase TERT expression compared to macronodules without TERT promoter mutations (P=0.004). Among 60 classical HCA of different molecular subtypes we didn't identified any mutations of the TERT promoter. In contrast, 7 among 16 HCA with malignant transformation (44 %) harbored TERT promoter mutations that were systematically associated with CTNNB1 mutations. Conclusion: TERT promoter mutations are the most frequent somatic genetic alterations observed in HCC. It is also the first recurrent somatic genetic alterations identified in cirrhotic preneoplastic macronodules suggesting that TERT promoter mutations is an early genetic event in the multistep process of cirrhotic carcinogene-sis. In contrast, TERT promoter mutations is not useful to promote initially benign liver tumorogenesis on normal liver but is required at the last step of malignant transformation of HCA in association with CTNNB1 mutations.
Jessica Zucman-Rossi - Consulting: pfizer; Grant/Research Support: Integragen; Speaking and Teaching: bayer, lilly
The following people have nothing to disclose: Jean-Charles Nault, Maxime Mallet, Camilla Pilati, Julien Calderaro, Paulette Bioulac-Sage, Christophe Laurent, Alexis Laurent, Daniel Cherqui, Charles Balabaud