Open AccessInvolvement of CD44 in induction of matrix metalloproteinases by a COOH‐terminal heparin‐binding fragment of fibronectin in human articular cartilage in culture
Author(s) -
Yasuda Tadashi,
Poole A. Robin,
Shimizu Makoto,
Nakagawa Takefumi,
Julovi Sohel M.,
Tamamura Hirokazu,
Fujii Nobutaka,
Nakamura Takashi
Publication year - 2003
Publication title -
arthritis & rheumatism
Language(s) - English
Resource type - Journals
eISSN - 1529-0131
pISSN - 0004-3591
DOI - 10.1002/art.10951
Subject(s) - collagenase , cartilage , matrix metalloproteinase , chemistry , proteoglycan , microbiology and biotechnology , fibronectin , peptide , type ii collagen , cd44 , interstitial collagenase , biochemistry , biology , extracellular matrix , in vitro , enzyme , anatomy
Abstract 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. Conclusion 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.