title: "Fibrous nests" in human hepatocellular carcinoma express a Wnt-induced gene signature associated with poor clinical outcome.
abstract: Hepatocellular carcinoma (HCC) is the 3rd cause of cancer-related death worldwide. Most cases arise in a background of chronic inflammation, extracellular matrix (ECM) remodeling, severe fibrosis and stem/progenitor cell amplification. Although HCCs are soft cellular tumors, they may contain fibrous nests within the tumor mass. Thus, the aim of this study was to explore cancer cell phenotypes in fibrous nests. Combined anatomic pathology, tissue microarray and real-time PCR analyses revealed that HCCs (n=82) containing fibrous nests were poorly differentiated, expressed Wnt pathway components and target genes, as well as markers of stem/progenitor cells, such as CD44, LGR5 and SOX9. Consistently, in severe liver fibroses (n=66) and in HCCs containing fibrous nests, weighted correlation analysis revealed a gene network including the myofibroblast marker ACTA2, the basement membrane components COL4A1 and LAMC1, the Wnt pathway members FZD1; FZD7; WNT2; LEF1; DKK1 and the Secreted Frizzled Related Proteins (SFRPs) 1; 2 and 5. Moreover, unbiased random survival forest analysis of a transcriptomic dataset of 247 HCC patients revealed high DKK1, COL4A1, SFRP1 and LAMC1 to be associated with advanced tumor staging as well as with bad overall and disease-free survival. In vitro, these genes were upregulated in liver cancer stem/progenitor cells upon Wnt-induced mesenchymal commitment and myofibroblast differentiation. In conclusion, fibrous nests express Wnt target genes, as well as markers of cancer stem cells and mesenchymal commitment. Fibrous nests embody the specific microenvironment of the cancer stem cell niche and can be detected by routine anatomic pathology analyses.
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France. Electronic address: rdesert87@gmail.com.
authors.forename: Romain
authors.initials: R
authors.lastname: Désert
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France. Electronic address: sihem.mebarki@gmail.com.
authors.forename: Sihem
authors.initials: S
authors.lastname: Mebarki
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France; CHU Rennes, Centre de Ressources Biologiques Santé BB-0033-00056, Rennes, France. Electronic address: mireille.desille-dugast@chu-rennes.fr.
authors.forename: Mireille
authors.initials: M
authors.lastname: Desille
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France. Electronic address: sicard.marie.i@gmail.com.
authors.forename: Marie
authors.initials: M
authors.lastname: Sicard
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France. Electronic address: elise.dessauge@gmail.com.
authors.forename: Elise
authors.initials: E
authors.lastname: Lavergne
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France. Electronic address: stephanie.renaud39@gmail.com.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Renaud
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France; CHU de Rennes, Dept. of Gastrointestinal and Hepatobiliary Surgery, Rennes, France. Electronic address: damien.bergeat@chu-rennes.fr.
authors.forename: Damien
authors.initials: D
authors.lastname: Bergeat
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France; CHU de Rennes, Dept. of Gastrointestinal and Hepatobiliary Surgery, Rennes, France. Electronic address: Laurent.Sulpice@chu-rennes.fr.
authors.forename: Laurent
authors.initials: L
authors.lastname: Sulpice
authors.affiliation: Inserm, U1016, Institut Cochin, Paris, France; Cnrs, UMR8104, Paris, France; Université Paris Descartes, Sorbonne Paris Cité, Paris, France. Electronic address: christine.perret@inserm.fr.
authors.forename: Christine
authors.initials: C
authors.lastname: Perret
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France; CHU Rennes, Centre de Ressources Biologiques Santé BB-0033-00056, Rennes, France. Electronic address: bruno.turlin@chu-rennes.fr.
authors.forename: Bruno
authors.initials: B
authors.lastname: Turlin
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France. Electronic address: bruno.clement@inserm.fr.
authors.forename: Bruno
authors.initials: B
authors.lastname: Clément
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France; Université de Rennes 1, F-35043 Rennes, France. Electronic address: orlando.musso@inserm.fr.
authors.forename: Orlando
authors.initials: O
authors.lastname: Musso
mesh_terms: D000199:Actins / Q000378:metabolism; D001485:Basement Membrane / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000150:complications / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology*; D002454:Cell Differentiation; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007797:Laminin / Q000378:metabolism; D008103:Liver Cirrhosis / Q000150:complications; D008113:Liver Neoplasms / Q000150:complications / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology*; D008648:Mesoderm / Q000473:pathology; D058628:Myofibroblasts / Q000473:pathology; D009361:Neoplasm Invasiveness; D014411:Neoplastic Stem Cells / Q000473:pathology; D011379:Prognosis; D012008:Recurrence; D059016:Tumor Microenvironment; D051153:Wnt Proteins / Q000378:metabolism*; D060449:Wnt Signaling Pathway / Q000235:genetics
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D014408:Biomarkers, Tumor; D007797:Laminin; D051153:Wnt Proteins; C089141:laminin gamma 1
keywords: CD44; Extracellular matrix remodeling; Fibrogenesis; Hepatocellular carcinoma; LGR5; SFRP; Stem cells; Transit-amplifying liver progenitor cells; Tumor microenvironment; Wnt; β-catenin