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

Romina Salpini, Matteo Surdo, Nadia Warner, Maria 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, Giuseppina Cappiello, Alberto Spanò, Pascale Trimoulet, Herve FleuryJacopo Vecchiet, Nerio Iapadre, Angelo Barlattani, Ada Bertoli, Terenzio Mari, Caterina Pasquazzi, Gabriele Missale, Cesare Sarrecchia, Elisa Orecchini, Alessandro Michienzi, Massimo Andreoni, Simona Francioso, Mario Angelico, Jens Verheyen, Francesca Ceccherini-Silberstein, Stephen Locarnini, Carlo Federico Perno, Valentina Svicher

Research output: Contribution to journalArticle

Abstract

Background: An impaired HBsAg-secretion can increase HBV oncogenicproperties. Here, we investigate genetic-determinants in HBsAg correlated with HBVinduced 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.

Original languageEnglish
Pages (from-to)15704-15715
Number of pages12
JournalOncotarget
Volume8
Issue number9
DOIs
Publication statusPublished - Jan 1 2017

Keywords

  • Cell proliferation
  • HBsAg mutations
  • Hepatitis B
  • Hepatitis B surface antigen
  • Hepatocellular carcinoma

ASJC Scopus subject areas

  • Oncology

Fingerprint Dive into the research topics of 'Novel HBsAg mutations correlate with hepatocellular carcinoma, hamper HBsAg secretion and promote cell proliferation in vitro'. Together they form a unique fingerprint.

  • Cite this

    Salpini, R., Surdo, M., Warner, N., Cortese, M. F., Colledge, D., Soppe, S., Bellocchi, M. C., Armenia, D., Carioti, L., Continenza, F., Di Carlo, D., Saccomandi, P., Mirabelli, C., Pollicita, M., Longo, R., Romano, S., Cappiello, G., Spanò, A., Trimoulet, P., ... Svicher, V. (2017). Novel HBsAg mutations correlate with hepatocellular carcinoma, hamper HBsAg secretion and promote cell proliferation in vitro. Oncotarget, 8(9), 15704-15715. https://doi.org/10.18632/oncotarget.14944