Portal de Periódicos da CAPES

Novel HBsAg mutations correlate with hepatocellular carcinoma, hamper HBsAg secretion and promote cell proliferation in vitro

2017; Impact Journals LLC; Volume: 8; Issue: 9 Linguagem: Inglês

10.18632/oncotarget.14944

1949-2553

Romina Salpini, Matteo Surdo, Nadia Warner, María Francesca Cortese, Danny Colledge, Sally Soppe, Maria Concetta Bellocchi, Daniele Armenia, Luca Carioti, Fabio Continenza, Domenico Di Carlo, Patrizia Saccomandi, Carmen Mirabelli, Michela Pollicita, Roberta Longo, Sara Romano-Bertrand, Giuseppina Cappiello, Alberto Spanò, Pascale Trimoulet, Hervé Fleury, Jacopo Vecchiet, N. Iapadre, A. Barlattani, Ada Bertoli, T. Mari, C. Pasquazzi, Gabriele Missale, C. Sarrecchia, Elisa Orecchini, Alessandro Michienzi, Massimo Andreoni, S. Francioso, M. Angélico, Jens Verheyen, Francesca Ceccherini‐Silberstein, Stephen Locarnini, Carlo Federico Perno, Valentina Svicher,

Liver Disease Diagnosis and Treatment

// Romina Salpini 1, * , Matteo Surdo 1, * , Nadia Warner 2 , Maria Francesca Cortese 1 , Danny Colledge 2 , Sally Soppe 2 , Maria Concetta Bellocchi 1 , Daniele Armenia 1 , Luca Carioti 1 , Fabio Continenza 3 , Domenico Di Carlo 1 , Patrizia Saccomandi 1 , Carmen Mirabelli 1, 4 , Michela Pollicita 1 , Roberta Longo 5 , Sara Romano 5 , Giuseppina Cappiello 5 , Alberto Spanò 5 , Pascale Trimoulet 6 , Herve Fleury 6 , Jacopo Vecchiet 7 , Nerio Iapadre 8 , Angelo Barlattani 9 , Ada Bertoli 1 , Terenzio Mari 10 , Caterina Pasquazzi 11 , Gabriele Missale 12 , Cesare Sarrecchia 13 , Elisa Orecchini 14 , Alessandro Michienzi 14 , Massimo Andreoni 13 , Simona Francioso 15 , Mario Angelico 15 , Jens Verheyen 16 , Francesca Ceccherini-Silberstein 1 , Stephen Locarnini 2 , Carlo Federico Perno 1 , Valentina Svicher 1  1 Department of Experimental Medicine and Surgery, University of Rome “Tor Vergata” Rome, Italy  2 Research and Molecular Development, Victorian Infectious Diseases Reference Laboratory, North Melbourne, Victoria, Australia  3 Laboratory of Monitoring Antiviral Drugs, National Institute for Infectious Diseases (INMI) “Lazzaro Spallanzani” Rome, Italy  4 Institut Pasteur, Unité de Biologie des Virus Entériques, Paris, France  5 Unit of Microbiology, “S. Pertini Hospital”, Rome, Italy  6 Laboratoire de Microbiologie Fondamentale et Pathogénicité, Hôpital Pellegrin Tripode, Bordeaux, France  7 Department of Medicine and Aging Sciences, “SS Annunziata” Hospital, Chieti, Italy  8 Infectious Diseases Unit, “S Salvatore” Hospital, L'Aquila, Italy  9 Hepatology Unit, “S Giacomo” Hospital, Rome, Italy 10 Hepatology Unit, “Regina Margherita” Hospital, Rome, Italy 11 Hepato-Infectivology Unit, “S Andrea” Hospital, Rome, Italy 12 Hospital of Parma, Parma, Italy 13 Tor Vergata University Hospital, Infectious Diseases Unit, Rome, Italy 14 Department of Biomedicine and Prevention, University of Rome “Tor Vergata” Rome, Italy 15 Tor Vergata University Hospital, Hepatology Unit, Rome, Italy 16 Institute of Virology, University Hospital, University of Duisburg-Essen, Essen, Germany * These authors have contributed equally to this work Correspondence to: Carlo Federico Perno, email: cf.perno@uniroma2.it Valentina Svicher, email: valentina.svicher@uniroma2.it Keywords: hepatitis B, hepatocellular carcinoma, hepatitis B surface antigen, HBsAg mutations, cell proliferation Received: November 25, 2016 Accepted: December 27, 2016 Published: February 01, 2017 ABSTRACT Background: An impaired HBsAg-secretion can increase HBV oncogenic-properties. Here, we investigate genetic-determinants in HBsAg correlated with HBV-induced hepatocellular carcinoma (HCC), and their impact on HBsAg-secretion and cell-proliferation. Methods: This study included 128 chronically HBV-infected patients: 23 with HCC (73.9% D; 26.1% A HBV-genotype), and 105 without cirrhosis/HCC (72.4% D, 27.6% A) as reference-group. The impact of mutations on HBsAg-secretion was assessed by measuring the ratio [secreted/intracellular HBsAg] until day 5 post-transfection. The impact of mutations on cell-cycle advancement was assessed by flow-cytometry. Results: Two HBsAg mutations significantly correlated with HCC: P203Q (17.4% [4/23] in HCC vs 1.0% [1/105] in non-HCC, P=0.004); S210R (34.8% [8/23] in HCC vs 3.8% [4/105] in non-HCC, P <0.001); P203Q+S210R (17.4% [4/23] in HCC vs 0% [0/110] in non-HCC, P=0.001). Both mutations reside in trans-membrane C-terminal domain critical for HBsAg-secretion. In in-vitro experiments, P203Q, S210R and P203Q+S210R significantly reduced the ratio [secreted/intracellular HBsAg] compared to wt at each time-point analysed (P <0.05), supporting an impaired HBsAg-secretion. Furthermore, P203Q and P203Q+S210R increased the percentage of cells in S-phase compared to wt, indicating cell-cycle progression (P203Q:26±13%; P203Q+S210R:29±14%; wt:18%±9, P <0.01. Additionally, S210R increased the percentage of cells in G2/M-phase (26±8% for wt versus 33±6% for S210R, P <0.001). Conclusions: Specific mutations in HBsAg C-terminus significantly correlate with HBV-induced HCC. They hamper HBsAg-secretion and are associated with increased cellular proliferation, supporting their involvement in HCC-development. The identification of viral genetic markers associated with HCC is critical to identify patients at higher HCC-risk that may deserve intensive liver monitoring, and/or early anti-HBV therapy.