title: Identification of opticin, a member of the small leucine-rich repeat proteoglycan family, in human articular tissues: a novel target for MMP-13 in osteoarthritis.
abstract: OBJECTIVE
  One of the proteoglycan families is the small leucine-rich proteoglycans (SLRPs) that are characterized by their association with collagen fibrils and/or some glycosaminoglycans. Opticin is a glycoprotein and class III member of the SLRP family, which was initially identified in the vitreous humour of the eye. In this study, we first investigated whether opticin is expressed and produced in normal and OA human articular tissues/cells. Further, we investigated the ability of the key metalloprotease involved in cartilage pathology, MMP-13, to cleave human cartilage opticin.
  
  
  METHODS
  Opticin gene expression was investigated in normal and OA human chondrocytes, synovial fibroblasts, and subchondral bone osteoblasts by reverse transcriptase-polymerase chain reaction (RT-PCR). Opticin protein production was determined in normal and OA synovial membrane and cartilage by immunohistochemistry. Opticin was isolated from human cartilage using guanidinium chloride extraction, and human MMP-13-induced opticin degradation analyzed by Western blotting. Finally, the opticin MMP-13 cleavage site was determined.
  
  
  RESULTS
  Opticin was expressed in human chondrocytes, synovial fibroblasts and subchondral osteoblasts, and the protein identified in synovial membrane and cartilage. At the protein level, OA cartilage showed a slightly higher level of opticin positive stained chondrocytes than normal cartilage; this did not reach statistical significance. However, in contrast with OA, normal cartilage demonstrated a high level of matrix staining in the superficial zone of the tissue, suggesting that in the OA cartilage matrix, opticin is degraded. Data also showed that cartilage opticin could be cleaved by MMP-13 after only 2h of incubation, indicating a preferential substrate compared to other SLRPs for this enzyme. Microsequencing revealed a major cleavage site at the G(104)/L(105)LAAP and a minor at P(109)/A(110)NHPG upon MMP-13 exposure.
  
  
  CONCLUSIONS
  We demonstrated, for the first time, that opticin is expressed and produced in human articular tissues. Our data also showed that opticin in OA cartilage is degraded in a process that could be mediated by MMP-13. As opticin may contribute towards the structural stability of cartilage, its cleavage by MMP-13 may predispose cartilage to degeneration, particularly at the surface.
authors.affiliation: Department of Rheumatology, Universitat Autonoma de Barcelona, Hospital del Mar, Passeig del Mar, Barcelona 08003, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Monfort
authors.forename: G
authors.initials: G
authors.lastname: Tardif
authors.forename: P
authors.initials: P
authors.lastname: Roughley
authors.forename: P
authors.initials: P
authors.lastname: Reboul
authors.forename: C
authors.initials: C
authors.lastname: Boileau
authors.forename: P N
authors.initials: PN
authors.lastname: Bishop
authors.forename: J-P
authors.initials: JP
authors.lastname: Pelletier
authors.forename: J
authors.initials: J
authors.lastname: Martel-Pelletier
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D019645:Arthroplasty, Replacement, Knee; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism; D019902:Chondrocytes / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007719:Knee Joint / Q000378:metabolism*; D053509:Matrix Metalloproteinase 13 / Q000494:pharmacology*; D008875:Middle Aged; D020370:Osteoarthritis, Knee / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D011509:Proteoglycans / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D013583:Synovial Membrane / Q000378:metabolism
chemical_list: D016326:Extracellular Matrix Proteins; C404108:OPTC protein, human; D011509:Proteoglycans; D012333:RNA, Messenger; D053509:Matrix Metalloproteinase 13