title: Fibroblast growth factor and canonical WNT/β-catenin signaling cooperate in suppression of chondrocyte differentiation in experimental models of FGFR signaling in cartilage.
abstract: Aberrant fibroblast growth factor (FGF) signaling disturbs chondrocyte differentiation in skeletal dysplasia, but the mechanisms underlying this process remain unclear. Recently, FGF was found to activate canonical WNT/β-catenin pathway in chondrocytes via Erk MAP kinase-mediated phosphorylation of WNT co-receptor Lrp6. Here, we explore the cellular consequences of such a signaling interaction. WNT enhanced the FGF-mediated suppression of chondrocyte differentiation in mouse limb bud micromass and limb organ cultures, leading to inhibition of cartilage nodule formation in micromass cultures, and suppression of growth in cultured limbs. Simultaneous activation of the FGF and WNT/β-catenin pathways resulted in loss of chondrocyte extracellular matrix, expression of genes typical for mineralized tissues and alteration of cellular shape. WNT enhanced the FGF-mediated downregulation of chondrocyte proteoglycan and collagen extracellular matrix via inhibition of matrix synthesis and induction of proteinases involved in matrix degradation. Expression of genes regulating RhoA GTPase pathway was induced by FGF in cooperation with WNT, and inhibition of the RhoA signaling rescued the FGF/WNT-mediated changes in chondrocyte cellular shape. Our results suggest that aberrant FGF signaling cooperates with WNT/β-catenin in suppression of chondrocyte differentiation.
authors.affiliation: Department of Anatomy, Histology and Embryology, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic; Institute of Animal Physiology and Genetics AS CR, v.v.i., Brno, Czech Republic.
authors.forename: Marcela
authors.initials: M
authors.lastname: Buchtova
authors.affiliation: Institute of Animal Physiology and Genetics AS CR, v.v.i., Brno, Czech Republic.
authors.forename: Veronika
authors.initials: V
authors.lastname: Oralova
authors.affiliation: Medical Genetics Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
authors.forename: Anie
authors.initials: A
authors.lastname: Aklian
authors.affiliation: Institute of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Masek
authors.affiliation: Department of Anatomy, Histology and Embryology, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Vesela
authors.affiliation: Department of Orthopaedics, Case Western Reserve University, Cleveland, OH, USA.
authors.forename: Zhufeng
authors.initials: Z
authors.lastname: Ouyang
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Tereza
authors.initials: T
authors.lastname: Obadalova
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Zaneta
authors.initials: Z
authors.lastname: Konecna
authors.affiliation: Institute of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic.
authors.forename: Tereza
authors.initials: T
authors.lastname: Spoustova
authors.affiliation: Institute of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic.
authors.forename: Tereza
authors.initials: T
authors.lastname: Pospisilova
authors.affiliation: Centre for Biomedical Image Analysis, Faculty of Informatics, Masaryk University, Brno, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Matula
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Varecha
authors.affiliation: Institute of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic.
authors.forename: Lukas
authors.initials: L
authors.lastname: Balek
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Gudernova
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Jelinkova
authors.affiliation: Department of Orthopaedic Surgery, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
authors.forename: Ivan
authors.initials: I
authors.lastname: Duran
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Iveta
authors.initials: I
authors.lastname: Cervenkova
authors.affiliation: Department of Orthopaedics, Case Western Reserve University, Cleveland, OH, USA.
authors.forename: Shunichi
authors.initials: S
authors.lastname: Murakami
authors.affiliation: Institute of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic; Department of Cytokinetics, Institute of Biophysics AS CR, v.v.i., Brno, Czech Republic.
authors.forename: Alois
authors.initials: A
authors.lastname: Kozubik
authors.affiliation: Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Dvorak
authors.affiliation: Institute of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic; Department of Cytokinetics, Institute of Biophysics AS CR, v.v.i., Brno, Czech Republic.
authors.forename: Vitezslav
authors.initials: V
authors.lastname: Bryja
authors.affiliation: Institute of Experimental Biology, Faculty of Sciences, Masaryk University, Brno, Czech Republic; Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic; International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic. Electronic address: krejcip@med.muni.cz.
authors.forename: Pavel
authors.initials: P
authors.lastname: Krejci
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002356:Cartilage / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism; D004357:Drug Synergism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005346:Fibroblast Growth Factors / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D018878:Limb Buds / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D060488:Low Density Lipoprotein Receptor-Related Protein-6 / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D051381:Rats; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D060509:Wnt3A Protein / Q000494:pharmacology; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism*
chemical_list: D060488:Low Density Lipoprotein Receptor-Related Protein-6; C555476:Lrp6 protein, mouse; D017468:Receptors, Fibroblast Growth Factor; D051153:Wnt Proteins; D060509:Wnt3A Protein; D051176:beta Catenin; D016222:Fibroblast Growth Factor 2; D005346:Fibroblast Growth Factors
keywords: Cartilage; Chondrocyte; Differentiation; FGFR3; Fibroblast growth factor receptor; WNT