Advertisement

Mixed hepatocellular cholangiocarcinoma tumors: Cholangiolocellular carcinoma is a distinct molecular entity

  • Agrin Moeini
    Affiliations
    Liver Cancer Translational Research Laboratory, Liver Unit, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, CIBERehd, Universitat de Barcelona, Barcelona, Catalonia, Spain

    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Daniela Sia
    Affiliations
    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Zhongyang Zhang
    Affiliations
    Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, USA

    Icahn Institute for Genomics and Multiscale Biology, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Genis Camprecios
    Affiliations
    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Ashley Stueck
    Affiliations
    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Hui Dong
    Affiliations
    Liver Cancer Translational Research Laboratory, Liver Unit, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, CIBERehd, Universitat de Barcelona, Barcelona, Catalonia, Spain
    Search for articles by this author
  • Robert Montal
    Affiliations
    Liver Cancer Translational Research Laboratory, Liver Unit, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, CIBERehd, Universitat de Barcelona, Barcelona, Catalonia, Spain
    Search for articles by this author
  • Laura Torrens
    Affiliations
    Liver Cancer Translational Research Laboratory, Liver Unit, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, CIBERehd, Universitat de Barcelona, Barcelona, Catalonia, Spain
    Search for articles by this author
  • Iris Martinez-Quetglas
    Affiliations
    Liver Cancer Translational Research Laboratory, Liver Unit, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, CIBERehd, Universitat de Barcelona, Barcelona, Catalonia, Spain
    Search for articles by this author
  • M. Isabel Fiel
    Affiliations
    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Ke Hao
    Affiliations
    Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, USA

    Icahn Institute for Genomics and Multiscale Biology, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Augusto Villanueva
    Affiliations
    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Swan N. Thung
    Affiliations
    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Myron E. Schwartz
    Affiliations
    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA
    Search for articles by this author
  • Josep M. Llovet
    Correspondence
    Corresponding author. Address: Director, Mount Sinai Liver Cancer Program, Division of Liver Diseases, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, Madison Ave 1425, 11F-70, Box: 1123, New York, NY 10029, USA. Tel.: +1 212 6599503; fax: +1 212 849 2574.
    Affiliations
    Liver Cancer Translational Research Laboratory, Liver Unit, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, CIBERehd, Universitat de Barcelona, Barcelona, Catalonia, Spain

    Mount Sinai Liver Cancer Program, (Divisions of Liver Diseases, Hematology and Medical Oncology, Department of Medicine, Department of Pathology, Recanati Miller Transplantation Institute), Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, USA

    Institució Catalana de Recerca i Estudis Avançats, Barcelona, Catalonia, Spain
    Search for articles by this author
Published:January 23, 2017DOI:https://doi.org/10.1016/j.jhep.2017.01.010

      Background & Aims

      Mixed hepatocellular cholangiocarcinoma (HCC-CCA) is a rare and poorly understood type of primary liver cancer. We aimed to perform a comprehensive molecular characterization of this malignancy.

      Methods

      Gene expression profiling, DNA copy number detection, and exome sequencing using formalin-fixed samples from 18 patients with mixed HCC-CCA were performed, encompassing the whole histological spectrum of the disease. Comparative genomic analysis was carried out, using independent datasets of HCC (n = 164) and intrahepatic cholangiocarcinoma (iCCA) (n = 149).

      Results

      Integrative genomic analysis of HCC-CCAs revealed that cholangiolocellular carcinoma (CLC) represents a distinct biliary-derived entity compared with the stem-cell and classical types. CLC tumors were neural cell adhesion molecule (NCAM) positive (6/6 vs. 1/12, p <0.001), chromosomally stable (mean chromosomal aberrations 5.7 vs. 14.1, p = 0.008), showed significant upregulation of transforming growth factor (TGF)-β signaling and enrichment of inflammation-related and immune response signatures (p <0.001). Stem-cell tumors were characterized by spalt-like transcription factor 4 (SALL4) positivity (6/8 vs. 0/10, p <0.001), enrichment of progenitor-like signatures, activation of specific oncogenic pathways (i.e., MYC and insulin-like growth factor [IGF]), and signatures related to poor clinical outcome. In the classical type, there was a significant correlation in the copy number variation of the iCCA and HCC components, suggesting a clonal origin. Exome sequencing revealed an average of 63 non-synonymous mutations per tumor (2 mean driver mutations per tumor). Among those, TP53 was the most frequently mutated gene (6/21, 29%) in HCC-CCAs.

      Conclusions

      Mixed HCC-CCA represents a heterogeneous group of tumors, with the stem-cell type characterized by features of poor prognosis, and the classical type with common lineage for HCC and iCCA components. CLC stands alone as a distinct biliary-derived entity associated with chromosomal stability and active TGF-β signaling.

      Lay summary

      Molecular analysis of mixed hepatocellular cholangiocarcinoma (HCC-CCA) showed that cholangiolocellular carcinoma (CLC) is distinct and biliary in origin. It has none of the traits of hepatocellular carcinoma (HCC). However, within mixed HCC-CCA, stem-cell type tumors shared an aggressive nature and poor outcome, whereas the classic type showed a common cell lineage for both the HCC and the intrahepatic CCA component. The pathological classification of mixed HCC-CCA should be redefined because of the new molecular data provided.

      Graphical abstract

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic and Personal
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Hepatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Llovet J.M.
        • Zucman-Rossi J.
        • Pikarsky E.
        • Sangro B.
        • Schwartz M.
        • Sherman M.
        • et al.
        Hepatocellular carcinoma.
        Nat Rev Dis Prim. 2016; 2: 16018
        • Theise N.D.
        • Nakashima O.
        • Park Y.N.
        • Nakanuma Y.
        Combined hepatocellular-cholangiocarcinoma.
        in: Bosman F.T. Carneiro F. Hruban R.H. Theise N.D. WHO classification of tumours of the digestive system. 4th ed. IARC, Lyons2010: 225-227
        • Jarnagin W.R.
        • Weber S.
        • Tickoo S.K.
        • Koea J.B.
        • Obiekwe S.
        • Fong Y.
        • et al.
        Combined hepatocellular and cholangiocarcinoma: demographic, clinical, and prognostic factors.
        Cancer. 2002; 94: 2040-2046
        • Wachtel M.S.
        • Zhang Y.
        • Xu T.
        • Chiriva-Internati M.
        • Frezza E.E.
        Combined hepatocellular cholangiocarcinomas; analysis of a large database.
        Clin Med Pathol. 2008; 1: 43-47
        • Sempoux C.
        • Fan C.
        • Singh P.
        • Obeidat K.
        • Roayaie S.
        • Schwartz M.
        • et al.
        Cholangiolocellular carcinoma: an innocent-looking malignant liver tumor mimicking ductular reaction.
        Semin Liver Dis. 2011; 31: 104-110
        • Lee J.-H.
        • Chung G.E.
        • Yu S.J.
        • Hwang S.Y.
        • Kim J.S.
        • Kim H.Y.
        • et al.
        Long-term prognosis of combined hepatocellular and cholangiocarcinoma after curative resection comparison with hepatocellular carcinoma and cholangiocarcinoma.
        J Clin Gastroenterol. 2011; 45: 69-75
        • Yin X.
        • Zhang B.-H.
        • Qiu S.-J.
        • Ren Z.-G.
        • Zhou J.
        • Chen X.-H.
        • et al.
        Combined hepatocellular carcinoma and cholangiocarcinoma: clinical features, treatment modalities, and prognosis.
        Ann Surg Oncol. 2012; 19: 2869-2876
        • Lee W.-S.
        • Lee K.-W.
        • Heo J.-S.
        • Kim S.-J.
        • Choi S.-H.
        • Kim Y.-I.
        • et al.
        Comparison of combined hepatocellular and cholangiocarcinoma with hepatocellular carcinoma and intrahepatic cholangiocarcinoma.
        Surg Today. 2006; 36: 892-897
        • Sasaki M.
        • Sato H.
        • Kakuda Y.
        • Sato Y.
        • Choi J.H.
        • Nakanuma Y.
        Clinicopathological significance of “subtypes with stem-cell feature” in combined hepatocellular-cholangiocarcinoma.
        Liver Int. 2015; 35: 1024-1035
        • Ikeda H.
        • Harada K.
        • Sato Y.
        • Sasaki M.
        • Yoneda N.
        • Kitamura S.
        • et al.
        Clinicopathologic significance of combined hepatocellular-cholangiocarcinoma with stem cell subtype components with reference to the expression of putative stem cell markers.
        Am J Clin Pathol. 2013; 140: 329-340
        • Komuta M.
        • Spee B.
        • Vander Borght S.
        • De Vos R.
        • Verslype C.
        • Aerts R.
        • et al.
        Clinicopathological study on cholangiolocellular carcinoma suggesting hepatic progenitor cell origin.
        Hepatology. 2008; 47: 1544-1556
        • Oishi N.
        • Kumar M.R.
        • Roessler S.
        • Ji J.
        • Forgues M.
        • Budhu A.
        • et al.
        Transcriptomic profiling reveals hepatic stem-like gene signatures and interplay of mir-200c and EMT in intrahepatic cholangiocarcinoma.
        Hepatology. 2012; 56: 1792-1803
        • Coulouarn C.
        • Cavard C.
        • Rubbia-Brandt L.
        • Audebourg A.
        • Dumont F.
        • Jacques S.
        • et al.
        Combined hepatocellular-cholangiocarcinomas exhibit progenitor features and activation of Wnt and TGFβ signaling pathways.
        Carcinogenesis. 2012; 33: 1791-1796
        • Fujimoto A.
        • Furuta M.
        • Shiraishi Y.
        • Gotoh K.
        • Kawakami Y.
        • Arihiro K.
        • et al.
        Whole-genome mutational landscape of liver cancers displaying biliary phenotype reveals hepatitis impact and molecular diversity.
        Nat Commun. 2015; 6: 6120
        • Woo H.G.
        • Lee J.H.
        • Yoon J.H.
        • Kim C.Y.
        • Lee H.S.
        • Jang J.J.
        • et al.
        Identification of a cholangiocarcinoma-like gene expression trait in hepatocellular carcinoma.
        Cancer Res. 2010; 70: 3034-3041
        • Cazals-Hatem D.
        • Rebouissou S.
        • Bioulac-Sage P.
        • Bluteau O.
        • Blanché H.
        • Franco D.
        • et al.
        Clinical and molecular analysis of combined hepatocellular-cholangiocarcinomas.
        J Hepatol. 2004; 41: 292-298
        • Villanueva A.
        • Hoshida Y.
        • Battiston C.
        • Tovar V.
        • Sia D.
        • Alsinet C.
        • et al.
        Combining clinical, pathology, and gene expression data to predict recurrence of hepatocellular carcinoma.
        Gastroenterology. 2011; 140: 1501-1512e2
        • Sia D.
        • Hoshida Y.
        • Villanueva A.
        • Roayaie S.
        • Ferrer J.
        • Tabak B.
        • et al.
        Integrative molecular analysis of intrahepatic cholangiocarcinoma reveals 2 classes that have different outcomes.
        Gastroenterology. 2013; 144: 829-840
        • Saha S.K.
        • Parachoniak C.A.
        • Ghanta K.S.
        • Fitamant J.
        • Ross K.N.
        • Najem M.S.
        • et al.
        Mutant IDH inhibits HNF-4α to block hepatocyte differentiation and promote biliary cancer.
        Nature. 2014; 513: 110-114
        • Hoshida Y.
        • Nijman S.M.B.
        • Kobayashi M.
        • Chan J.A.
        • Brunet J.-P.P.
        • Chiang D.Y.
        • et al.
        Integrative transcriptome analysis reveals common molecular subclasses of human hepatocellular carcinoma.
        Cancer Res. 2009; 69: 7385-7392
        • Coulouarn C.
        • Factor V.M.
        • Thorgeirsson S.S.
        Transforming growth factor-beta gene expression signature in mouse hepatocytes predicts clinical outcome in human cancer.
        Hepatology. 2008; 47: 2059-2067
        • Bindea G.
        • Mlecnik B.
        • Tosolini M.
        • Kirilovsky A.
        • Waldner M.
        • Obenauf A.C.
        • et al.
        Spatiotemporal dynamics of intratumoral immune cells reveal the immune landscape in human cancer.
        Immunity. 2013; 39: 782-795
        • Yamashita T.
        • Forgues M.
        • Wang W.
        • Kim J.W.
        • Ye Q.
        • Jia H.
        • et al.
        EpCAM and alpha-fetoprotein expression defines novel prognostic subtypes of hepatocellular carcinoma.
        Cancer Res. 2008; 68: 1451-1461
        • Chiang D.Y.Y.
        • Villanueva A.
        • Hoshida Y.
        • Peix J.
        • Newell P.
        • Minguez B.
        • et al.
        Focal gains of VEGFA and molecular classification of hepatocellular carcinoma.
        Cancer Res. 2008; 68: 6779-6788
        • Boyault S.
        • Rickman D.S.
        • de Reyniès A.
        • Balabaud C.
        • Rebouissou S.
        • Jeannot E.
        • et al.
        Transcriptome classification of HCC is related to gene alterations and to new therapeutic targets.
        Hepatology. 2007; 45: 42-52
        • Lee J.-S.
        • Chu I.-S.
        • Heo J.
        • Calvisi D.F.
        • Sun Z.
        • Roskams T.
        • et al.
        Classification and prediction of survival in hepatocellular carcinoma by gene expression profiling.
        Hepatology. 2004; 40: 667-676
        • Tovar V.
        • Alsinet C.
        • Villanueva A.
        • Hoshida Y.
        • Chiang D.Y.Y.
        • Solé M.
        • et al.
        IGF activation in a molecular subclass of hepatocellular carcinoma and pre-clinical efficacy of IGF-1R blockage.
        J Hepatol. 2010; 52: 550-559
        • Villanueva A.
        • Alsinet C.
        • Yanger K.
        • Hoshida Y.
        • Zong Y.
        • Toffanin S.
        • et al.
        Notch signaling is activated in human hepatocellular carcinoma and induces tumor formation in mice.
        Gastroenterology. 2012; 143: 1660-1669e7
        • Andersen J.B.
        • Spee B.
        • Blechacz B.R.
        • Avital I.
        • Komuta M.
        • Barbour A.
        • et al.
        Genomic and genetic characterization of cholangiocarcinoma identifies therapeutic targets for tyrosine kinase inhibitors.
        Gastroenterology. 2012; 142: 1021-1031e15
        • Sia D.
        • Losic B.
        • Moeini A.
        • Cabellos L.
        • Hao K.
        • Revill K.
        • et al.
        Massive parallel sequencing uncovers actionable FGFR2-PPHLN1 fusion and ARAF mutations in intrahepatic cholangiocarcinoma.
        Nat Commun. 2015; 6: 6087
        • Ong C.K.
        • Subimerb C.
        • Pairojkul C.
        • Wongkham S.
        • Cutcutache I.
        • Yu W.
        • et al.
        Exome sequencing of liver fluke-associated cholangiocarcinoma.
        Nat Genet. 2012; 44: 690-693
        • Schulze K.
        • Imbeaud S.
        • Letouzé E.
        • Alexandrov L.B.
        • Calderaro J.
        • Rebouissou S.
        • et al.
        Exome sequencing of hepatocellular carcinomas identifies new mutational signatures and potential therapeutic targets.
        Nat Genet. 2015; 47: 505-511
        • Totoki Y.
        • Tatsuno K.
        • Covington K.R.
        • Ueda H.
        • Creighton C.J.
        • Kato M.
        • et al.
        Trans-ancestry mutational landscape of hepatocellular carcinoma genomes.
        Nat Genet. 2014; 46: 1267-1273
        • Greenman C.
        • Stephens P.
        • Smith R.
        • Dalgliesh G.L.
        • Hunter C.
        • Bignell G.
        • et al.
        Patterns of somatic mutation in human cancer genomes.
        Nature. 2007; 446: 153-158
        • Garraway L.A.
        • Lander E.S.
        Lessons from the cancer genome.
        Cell. 2013; 153: 17-37
        • Vogelstein B.
        • Papadopoulos N.
        • Velculescu V.E.
        • Zhou S.
        • Diaz L.A.
        • Kinzler K.W.
        Cancer genome landscapes.
        Science. 2013; 339: 1546-1558
        • Hoshida Y.
        • Brunet J.-P.
        • Tamayo P.
        • Golub T.R.
        • Mesirov J.P.
        Subclass mapping: identifying common subtypes in independent disease data sets.
        PLoS One. 2007; 2e1195
        • Komuta M.
        • Govaere O.
        • Vandecaveye V.
        • Akiba J.
        • Van Steenbergen W.
        • Verslype C.
        • et al.
        Histological diversity in cholangiocellular carcinoma reflects the different cholangiocyte phenotypes.
        Hepatology. 2012; 55: 1876-1888
        • Akiba J.
        • Nakashima O.
        • Hattori S.
        • Tanikawa K.
        • Takenaka M.
        • Nakayama M.
        • et al.
        Clinicopathologic analysis of combined hepatocellular-cholangiocarcinoma according to the latest WHO classification.
        Am J Surg Pathol. 2013; 37: 496-505
        • Massagué J.
        TGFbeta in Cancer.
        Cell. 2008; 134: 215-230
        • Wu K.
        • Ding J.
        • Chen C.
        • Sun W.
        • Ning B.-F.
        • Wen W.
        • et al.
        Hepatic transforming growth factor beta gives rise to tumor-initiating cells and promotes liver cancer development.
        Hepatology. 2012; 56: 2255-2267
        • Karkampouna S.
        • Ten Dijke P.
        • Dooley S.
        • Julio M.K.
        TGFβ signaling in liver regeneration.
        Curr Pharm Des. 2012; 18: 4103-4113
        • Chen Y.
        • Huang Y.
        • Reiberger T.
        • Duyverman A.M.
        • Huang P.
        • Samuel R.
        • et al.
        Differential effects of sorafenib on liver vs. tumor fibrosis mediated by stromal-derived factor 1 alpha/C-X-C receptor type 4 axis and myeloid differentiation antigen-positive myeloid cell infiltration in mice.
        Hepatology. 2014; 59: 1435-1447
        • Chew V.
        • Chen J.
        • Lee D.
        • Loh E.
        • Lee J.
        • Lim K.H.
        • et al.
        Chemokine-driven lymphocyte infiltration: an early intratumoural event determining long-term survival in resectable hepatocellular carcinoma.
        Gut. 2012; 61: 427-438
        • Gao Q.
        • Qiu S.-J.
        • Fan J.
        • Zhou J.
        • Wang X.-Y.
        • Xiao Y.-S.
        • et al.
        Intratumoral balance of regulatory and cytotoxic T cells is associated with prognosis of hepatocellular carcinoma after resection.
        J Clin Oncol. 2007; 25: 2586-2593
        • Galon J.
        • Costes A.
        • Sanchez-Cabo F.
        • Kirilovsky A.
        • Mlecnik B.
        • Lagorce-Pagès C.
        • et al.
        Type, density, and location of immune cells within human colorectal tumors predict clinical outcome.
        Science. 2006; 313: 1960-1964
        • Yong K.J.
        • Gao C.
        • Lim J.S.J.
        • Yan B.
        • Yang H.
        • Dimitrov T.
        • et al.
        Oncofetal gene SALL4 in aggressive hepatocellular carcinoma.
        N Engl J Med. 2013; 368: 2266-2276
        • Fujii H.
        • Zhu X.G.
        • Matsumoto T.
        • Inagaki M.
        • Tokusashi Y.
        • Miyokawa N.
        • et al.
        Genetic classification of combined hepatocellular-cholangiocarcinoma.
        Hum Pathol. 2000; 31: 1011-1017
        • Moeini A.
        • Sia D.
        • Bardeesy N.
        • Mazzaferro V.
        • Llovet J.M.
        Molecular pathogenesis and targeted therapies for intrahepatic cholangiocarcinoma.
        Clin Cancer Res. 2016; 22: 291-300