HBV-related HCC development in mice is STAT3 dependent and indicates an oncogenic effect of HBx
Marc Ringelhan,
Svenja Schuehle,
Maarten van de Klundert,
Elena Kotsiliti,
Marie-Laure Plissonnier,
Suzanne Faure-Dupuy,
Tobias Riedl,
Sebastian Lange,
Karin Wisskirchen,
Frank Thiele,
Cho-Chin Cheng,
Detian Yuan,
Valentina Leone,
Ronny Schmidt,
Juliana Hünergard,
Fabian Geisler,
Kristian Unger,
Hana Algül,
Roland M. Schmid,
Roland Rad,
Heiner Wedemeyer,
Massimo Levrero,
Ulrike Protzer,
Mathias Heikenwalder
Affiliations
Marc Ringelhan
Second Medical Department, University Hospital Rechts der Isar, Technical University of Munich, School of Medicine & Health, Munich, Germany; German Centre for Infection Research (DZIF), Munich Partner Site, Munich, Germany
Svenja Schuehle
German Cancer Research Center (DKFZ), Heidelberg, Germany; Faculty of Biosciences, Heidelberg University, Heidelberg, Germany
Maarten van de Klundert
Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany
Elena Kotsiliti
German Cancer Research Center (DKFZ), Heidelberg, Germany; Faculty of Biosciences, Heidelberg University, Heidelberg, Germany
Marie-Laure Plissonnier
University of Lyon, Lyon, France; Inserm U1052, Cancer Research Centre of Lyon, Lyon, France
Suzanne Faure-Dupuy
German Cancer Research Center (DKFZ), Heidelberg, Germany
Tobias Riedl
German Cancer Research Center (DKFZ), Heidelberg, Germany
Sebastian Lange
Second Medical Department, University Hospital Rechts der Isar, Technical University of Munich, School of Medicine & Health, Munich, Germany; Institute of Molecular Oncology and Functional Genomics, School of Medicine, Technical University of Munich, Munich, Germany; Center for Translational Cancer Research (TranslaTUM), School of Medicine & Health, Technical University of Munich, Munich, Germany; German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany
Karin Wisskirchen
Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany
Frank Thiele
Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany
Cho-Chin Cheng
Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany
Detian Yuan
Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany
Valentina Leone
German Cancer Research Center (DKFZ), Heidelberg, Germany; Research Unit for Radiation Cytogenetics, Helmholtz Munich, Neuherberg, Germany
Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany
Fabian Geisler
Second Medical Department, University Hospital Rechts der Isar, Technical University of Munich, School of Medicine & Health, Munich, Germany
Kristian Unger
Research Unit for Radiation Cytogenetics, Helmholtz Munich, Neuherberg, Germany; Department of Radiation Oncology, University Hospital, LMU Munich, Munich, Germany
Hana Algül
Second Medical Department, University Hospital Rechts der Isar, Technical University of Munich, School of Medicine & Health, Munich, Germany; Comprehensive Cancer Center TUM (CCCMTUM), University Hospital rechts der Isar, Technical University of Munich, School of Medicine & Health, Munich, Germany
Roland M. Schmid
Second Medical Department, University Hospital Rechts der Isar, Technical University of Munich, School of Medicine & Health, Munich, Germany
Roland Rad
Second Medical Department, University Hospital Rechts der Isar, Technical University of Munich, School of Medicine & Health, Munich, Germany; Institute of Molecular Oncology and Functional Genomics, School of Medicine, Technical University of Munich, Munich, Germany; Center for Translational Cancer Research (TranslaTUM), School of Medicine & Health, Technical University of Munich, Munich, Germany; German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany
Heiner Wedemeyer
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany; German Center for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Germany
Massimo Levrero
INSERM Unit 1052, Cancer Research Center of Lyon, Lyon, France; Hepatology Department, Hospices Civils de Lyon, Lyon, France; Department of Internal Medicine - DMISM, Sapienza University, Rome, Italy; Istituto Italiano di Tecnologia (IIT), Rome, Italy
Ulrike Protzer
German Centre for Infection Research (DZIF), Munich Partner Site, Munich, Germany; Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany; Corresponding authors. Addresses: Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany (U. Protzer), The M3 Research Center, Medical Faculty, University Tubingen, Tubingen, Germany (M. Heikenwalder).
Mathias Heikenwalder
German Cancer Research Center (DKFZ), Heidelberg, Germany; Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany; The M3 Research Center, Medical Faculty, University Tübingen, Tübingen, Germany; Cluster of Excellence iFIT (EXC 2180) “Image-Guided and Functionally Instructed Tumor Therapies”, University of Tübingen, Tübingen, Germany; Department of Infectious Diseases, Molecular Virology, Heidelberg University, Heidelberg, Germany; Corresponding authors. Addresses: Institute of Virology, Technical University of Munich, School of Medicine & Health/Helmholtz Munich, Munich, Germany (U. Protzer), The M3 Research Center, Medical Faculty, University Tubingen, Tubingen, Germany (M. Heikenwalder).
Background & Aims: Although most hepatocellular carcinoma (HCC) cases are driven by hepatitis and cirrhosis, a subset of patients with chronic hepatitis B develop HCC in the absence of advanced liver disease, indicating the oncogenic potential of hepatitis B virus (HBV). We investigated the role of HBV transcripts and proteins on HCC development in the absence of inflammation in HBV-transgenic mice. Methods: HBV-transgenic mice replicating HBV and expressing all HBV proteins from a single integrated 1.3-fold HBV genome in the presence or absence of wild-type HBx (HBV1.3/HBVxfs) were analyzed. Flow cytometry, molecular, histological and in vitro analyses using human cell lines were performed. Hepatocyte-specific Stat3- and Socs3-knockout was analyzed in HBV1.3 mice. Results: Approximately 38% of HBV1.3 mice developed liver tumors. Protein expression patterns, histology, and mutational landscape analyses indicated that tumors resembled human HCC. HBV1.3 mice showed no signs of active hepatitis, except STAT3 activation, up to the time point of HCC development. HBV-RNAs covering HBx sequence, 3.5-kb HBV RNA and HBx-protein were detected in HCC tissue. Interestingly, HBVxfs mice expressing all HBV proteins except a C-terminally truncated HBx (without the ability to bind DNA damage binding protein 1) showed reduced signs of DNA damage response and had a significantly reduced HCC incidence. Importantly, intercrossing HBV1.3 mice with a hepatocyte-specific STAT3-knockout abrogated HCC development. Conclusions: Expression of HBV-proteins is sufficient to cause HCC in the absence of detectable inflammation. This indicates the oncogenic potential of HBV and in particular HBx. In our model, HBV-driven HCC was STAT3 dependent. Our study highlights the immediate oncogenic potential of HBV, challenging the idea of a benign highly replicative phase of HBV infection and indicating the necessity for an HBV ‘cure’. Impact and implications: Although most HCC cases in patients with chronic HBV infection occur after a sequence of liver damage and fibrosis, a subset of patients develops HCC without any signs of advanced liver damage. We demonstrate that the expression of all viral transcripts in HBV-transgenic mice suffices to induce HCC development independent of inflammation and fibrosis. These data indicate the direct oncogenic effects of HBV and emphasize the idea of early antiviral treatment in the ‘immune-tolerant’ phase (HBeAg-positive chronic HBV infection).