title: De novo HAPLN1 expression hallmarks Wnt-induced stem cell and fibrogenic networks leading to aggressive human hepatocellular carcinomas.
abstract: About 20% hepatocellular carcinomas (HCCs) display wild-type β-catenin, enhanced Wnt signaling, hepatocyte dedifferentiation and bad outcome, suggesting a specific impact of Wnt signals on HCC stem/progenitor cells. To study Wnt-specific molecular pathways, cell fates and clinical outcome, we fine-tuned Wnt/β-catenin signaling in liver progenitor cells, using the prototypical Wnt ligand Wnt3a. Cell biology assays and transcriptomic profiling were performed in HepaRG hepatic progenitors exposed to Wnt3a after β-catenin knockdown or Wnt inhibition with FZD8_CRD. Gene expression network, molecular pathology and survival analyses were performed on HCCs and matching non-tumor livers from 70 patients by real-time PCR and tissue micro-array-based immunohistochemistry. Wnt3a reprogrammed liver progenitors to replicating fibrogenic myofibroblast-like cells displaying stem and invasive features. Invasion was inhibited by 30 nM FZD7 and FZD8 CRDs. Translation of these data to human HCCs revealed two tight gene networks associating cell surface Wnt signaling, stem/progenitor markers and mesenchymal commitment. Both networks were linked by Hyaluronan And Proteoglycan Link Protein 1 (HAPLN1), that appeared de novo in aggressive HCCs expressing cytoplasmic β-catenin and stem cell markers. HAPLN1 was independently associated with bad overall and disease-free outcome. In vitro, HAPLN1 was expressed de novo in EPCAM¯/NCAM+ mesoderm-committed progenitors, upon spontaneous epithelial-mesenchymal transition and de-differentiation of hepatocyte-like cells to liver progenitors. In these cells, HAPLN1 knockdown downregulated key markers of mesenchymal cells, such as Snail, LGR5, collagen IV and α-SMA. In conclusion, HAPLN1 reflects a signaling network leading to stemness, mesenchymal commitment and HCC progression.
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Sihem
authors.initials: S
authors.lastname: Mebarki
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Romain
authors.initials: R
authors.lastname: Désert
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.|CHU de Rennes, Department of Gastrointestinal and Hepatobiliary Surgery, Rennes, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Sulpice
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Sicard
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.|CHU de Rennes, Centre de Ressources Biologiques Santé, Rennes, France.
authors.forename: Mireille
authors.initials: M
authors.lastname: Desille
authors.affiliation: Inserm, Institut Cochin, Paris, France.|Cnrs, UMR8104, Paris, France.|Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Canal
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Dubois-Pot Schneider
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.|CHU de Rennes, Department of Gastrointestinal and Hepatobiliary Surgery, Rennes, France.
authors.forename: Damien
authors.initials: D
authors.lastname: Bergeat
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.|CHU de Rennes, Centre de Ressources Biologiques Santé, Rennes, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Turlin
authors.affiliation: Université de Rennes 1, UMS 18 Biosit, Biogenouest, Rennes, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Bellaud
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Elise
authors.initials: E
authors.lastname: Lavergne
authors.affiliation: Université de Rennes 1, UMS 18 Biosit, Biogenouest, ImPACcellCore Facility, Rennes, France.
authors.forename: Rémy
authors.initials: R
authors.lastname: Le Guével
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.|Université de Rennes 1, UMS 18 Biosit, Biogenouest, ImPACcellCore Facility, Rennes, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Corlu
authors.affiliation: Inserm, Institut Cochin, Paris, France.|Cnrs, UMR8104, Paris, France.|Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Perret
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Coulouarn
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Clément
authors.affiliation: Inserm, UMR991, Liver Metabolisms and Cancer, Rennes, France.|Université de Rennes 1, Rennes, France.
authors.forename: Orlando
authors.initials: O
authors.lastname: Musso
mesh_terms: D000368:Aged; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005500:Follow-Up Studies; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008024:Ligands; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008113:Liver Neoplasms / Q000378:metabolism*; D008297:Male; D008648:Mesoderm / Q000378:metabolism*; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D046888:Tissue Array Analysis; D051153:Wnt Proteins / Q000378:metabolism*; D000099249:Proteoglycan Link Protein
chemical_list: D016326:Extracellular Matrix Proteins; D008024:Ligands; D011509:Proteoglycans; D051153:Wnt Proteins; D000099249:Proteoglycan Link Protein
keywords: CD44; Gerotarget; LGR5; Wnt; epithelial-mesenchymal transition; β-catenin