Involvement of CD44 in induction of matrix metalloproteinases by a COOH-terminal heparin-binding fragment of fibronectin in human articular cartilage in culture.
OBJECTIVE
To investigate the mechanism of induction of matrix metalloproteinases (MMPs) by a 40-kd COOH-terminal heparin-binding fibronectin fragment (HBFN-f) containing III12-14 and IIICS domains in human articular cartilage in culture.


METHODS
Human articular cartilage was removed from macroscopically normal femoral heads and cultured with HBFN-f. MMP secretion into conditioned media was analyzed by immunoblotting (MMPs 1 and 13) and by gelatin zymography (MMPs 2 and 9). Type II collagen cleavage by collagenase was monitored in culture by immunoassay. Involvement of specific peptide-binding domains in HBFN-f and the involvement of CD44 were assessed with synthetic peptides and an anti-CD44 antibody. Immunofluorescence histochemistry was performed using fluorescein isothiocyanate-conjugated anti-CD44 antibody.


RESULTS
HBFN-f stimulated production of MMPs 1, 2, 9, and 13 in association with type II collagen cleavage by collagenase in human articular cartilage. Peptide V (WQPPRARI) of HBFN-f, which can bind cell surface heparan sulfate proteoglycan (HSPG), blocked MMP induction by HBFN-f, while the scrambled peptide V (RPQIPWAR) had no effect. Peptide CS-1 of 25 amino acids in IIICS of HBFN-f caused no significant effect. Treatment of cartilage with anti-CD44 antibody or HSPG resulted in significant inhibition of HBFN-f-stimulated MMP production. Preincubation with peptide V blocked binding of the anti-CD44 antibody to chondrocytes in cartilage.


CONCLUSIONS
Interaction of the peptide V sequence in HBFN-f with glycosaminoglycans, such as those in CD44, plays an important role in HBFN-f-stimulated MMP production in articular cartilage. Because CD44 is up-regulated in osteoarthritic and rheumatoid arthritic cartilage, the role of the interaction between CD44 and HBFN-f in these pathologies should be of relevance and should be studied further.
Kyoto University Graduate School of Medicine, Kyoto, Japan. tadyasu@kuhp.kyoto-u.ac.jp
Tadashi
T
Yasuda
A Robin
AR
Poole
Makoto
M
Shimizu
Takefumi
T
Nakagawa
Sohel M
SM
Julovi
Hirokazu
H
Tamamura
Nobutaka
N
Fujii
Takashi
T
Nakamura
D002352:Carrier Proteins / Q000494:pharmacology; D002358:Cartilage, Articular / Q000187:drug effects* / Q000201:enzymology; D002478:Cells, Cultured; D024043:Collagen Type II / Q000378:metabolism; D017364:Collagenases / Q000096:biosynthesis*; D004247:DNA / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction / Q000187:drug effects; D005353:Fibronectins / Q000378:metabolism / Q000494:pharmacology*; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology; D019085:Fluorescent Antibody Technique, Direct; D019812:Heparan Sulfate Proteoglycans / Q000494:pharmacology; D006493:Heparin / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000276:immunology / Q000378:metabolism*; D015151:Immunoblotting; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis*; D053509:Matrix Metalloproteinase 13; D009479:Neuropeptides / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology
D002352:Carrier Proteins; D024043:Collagen Type II; D005353:Fibronectins; D019812:Heparan Sulfate Proteoglycans; D018960:Hyaluronan Receptors; D009479:Neuropeptides; D009842:Oligopeptides; D010446:Peptide Fragments; C094697:glutamyl-isoleucyl-leucyl-aspartyl-valine; C090783:peptide V; D006493:Heparin; D004247:DNA; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D020781:Matrix Metalloproteinase 1; D016650:Fluorescein-5-isothiocyanate