Model of fibrolamellar hepatocellular carcinomas reveals striking enrichment in cancer stem cells

Tsunekazu Oikawa; Eliane Wauthier; Timothy A Dinh; Sara R Selitsky; Andrea Reyna-Neyra; Guido Carpino; Ronald Levine; Vincenzo Cardinale; David Klimstra; Eugenio Gaudio; Domenico Alvaro; Nancy Carrasco; Praveen Sethupathy; Lola M Reid

doi:10.1038/ncomms9070

Model of fibrolamellar hepatocellular carcinomas reveals striking enrichment in cancer stem cells

Nat Commun. 2015 Oct 6:6:8070. doi: 10.1038/ncomms9070.

Authors

Tsunekazu Oikawa^{1

2

3}, Eliane Wauthier^{1

2

3}, Timothy A Dinh^{4

5

6}, Sara R Selitsky^{4

7}, Andrea Reyna-Neyra⁸, Guido Carpino⁹, Ronald Levine^{10

11}, Vincenzo Cardinale¹², David Klimstra¹³, Eugenio Gaudio¹⁴, Domenico Alvaro¹², Nancy Carrasco⁸, Praveen Sethupathy^{3

4

5

7}, Lola M Reid^{1

2

3}

Affiliations

¹ Department of Cell Biology and Physiology, Chapel Hill, North Carolina 27599, USA.
² Program in Molecular Biology and Biotechnology, Chapel Hill, North Carolina 27599, USA.
³ Lineberger Comprehensive Cancer Center, Chapel Hill, North Carolina 27599, USA.
⁴ Department of Genetics, Chapel Hill, North Carolina 27599, USA.
⁵ Curriculum in Genetics and Molecular Biology, Chapel Hill, North Carolina 27599, USA.
⁶ MD-PhD Program, UNC School of Medicine, Chapel Hill, North Carolina 27599, USA.
⁷ Curriculum in Bioinformatics and Computational Biology, UNC School of Medicine, Chapel Hill, North Carolina 27599, USA.
⁸ Department of Cellular and Molecular Physiology, Yale University School of Medicine, 333 Cedar Street, New Haven, Connecticut 06510, USA.
⁹ Department of Movement, Human and Health Sciences, Division of Health Sciences, University of Rome 'Foro Italico', Piazza Lauro De Bosis 6, 00151 Rome, Italy.
¹⁰ Greenwich Hospital, 5 Perryridge Road, Greenwich, Connecticut 06830, USA.
¹¹ Yale University School of Medicine, 333 Cedar Street, New Haven, Connecticut, 06510 USA.
¹² Division of Gastroenterology, Department of Scienze e Biotecnologie Medico-Chirurgiche, Fondazione Eleonora Lorillard Spencer Cenci, Polo Pontino, Viale dell'Universita 37, 00185 Rome, Italy.
¹³ Department of Pathology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, New York 10065, USA.
¹⁴ Department of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University of Rome, Via Borelli 50, 00161 Rome, Italy.

Abstract

The aetiology of human fibrolamellar hepatocellular carcinomas (hFL-HCCs), cancers occurring increasingly in children to young adults, is poorly understood. We present a transplantable tumour line, maintained in immune-compromised mice, and validate it as a bona fide model of hFL-HCCs by multiple methods. RNA-seq analysis confirms the presence of a fusion transcript (DNAJB1-PRKACA) characteristic of hFL-HCC tumours. The hFL-HCC tumour line is highly enriched for cancer stem cells as indicated by limited dilution tumourigenicity assays, spheroid formation and flow cytometry. Immunohistochemistry on the hFL-HCC model, with parallel studies on 27 primary hFL-HCC tumours, provides robust evidence for expression of endodermal stem cell traits. Transcriptomic analyses of the tumour line and of multiple, normal hepatic lineage stages reveal a gene signature for hFL-HCCs closely resembling that of biliary tree stem cells--newly discovered precursors for liver and pancreas. This model offers unprecedented opportunities to investigate mechanisms underlying hFL-HCCs pathogenesis and potential therapies.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Adolescent
Adult
Animals
Biomarkers, Tumor / metabolism*
Carcinoma, Hepatocellular / genetics*
Carcinoma, Hepatocellular / metabolism
Carcinoma, Hepatocellular / pathology
Cell Line, Tumor
Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / genetics
Female
Flow Cytometry
Gene Expression Profiling
Gene Expression Regulation, Neoplastic*
HSP40 Heat-Shock Proteins / genetics
Humans
Immunohistochemistry
Liver Neoplasms / genetics*
Liver Neoplasms / metabolism
Liver Neoplasms / pathology
Male
Mice
Mice, Nude
Mice, SCID
Microscopy, Electron, Transmission
Middle Aged
Mutant Chimeric Proteins / genetics*
Neoplasm Transplantation
Neoplastic Stem Cells / metabolism*
RNA, Messenger / metabolism*
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, RNA
Spheroids, Cellular
Tumor Cells, Cultured
Young Adult

Substances

Biomarkers, Tumor
DNAJB1 protein, human
HSP40 Heat-Shock Proteins
Mutant Chimeric Proteins
RNA, Messenger
Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
PRKACA protein, human

Model of fibrolamellar hepatocellular carcinomas reveals striking enrichment in cancer stem cells

Authors

Affiliations

Abstract

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Substances

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