Activation of the Discoidin Domain Receptor 2 Induces Expression of Matrix Metalloproteinase 13 Associated with Osteoarthritis in Mice*♦

Human genetic studies indicate that mutations in type IX and XI collagens result in early-onset osteoarthritis (OA) with a wide spectrum of osteochondrodysplasia. However, a convincing causal chain of events underlying the role of these collagen mutations in the pathogenesis of OA has not been elucidated. Here we show that the expression of a cell surface collagen receptor, discoidin domain receptor 2 (DDR2), is increased in chondrocytes of the articular cartilage of knee joints in mice that develop OA as a result of a heterozygous mutation in type XI collagen. At the same time point, 6 months, we also found increased expression and activity of matrix metalloproteinase 13 (MMP-13) in the mutant mouse knee cartilage. The expression of both DDR2 and MMP-13 was increased in chondrocytes cultured on plates coated with native type II collagen but not on gelatin, and overexpression of DDR2, but not of a truncated form, was found to induce the expression of MMP-13 when chondrocytes were cultured on type II collagen but not on plastic. The DDR2-induced expression of MMP-13 appears to be specific, since we did not observe induction of MMP-1, MMP-3, MMP-8, ADAMTS-4, ADAMTS-5, and IL-1 transcripts in human chondrocytes or Mmp-3, Mmp-8, Adamts-4, Adamts-5, and Il-1 in mouse chondrocytes. Our data suggest that the defect in the cartilage matrix of mice that are heterozygous for a type XI collagen mutation (cho/+) permits activation and up-regulation of DDR2 in chondrocytes. This could be due to increased exposure of chondrocytes to type II collagen as a result of the decreased amount of type XI collagen in the mutant cartilage. The specific induction of MMP-13 by DDR2 in response to its cartilage-specific ligand, type II collagen, may contribute to cartilage damage in hereditary OA. Human genetic studies indicate that mutations in type IX and XI collagens result in early-onset osteoarthritis (OA) with a wide spectrum of osteochondrodysplasia. However, a convincing causal chain of events underlying the role of these collagen mutations in the pathogenesis of OA has not been elucidated. Here we show that the expression of a cell surface collagen receptor, discoidin domain receptor 2 (DDR2), is increased in chondrocytes of the articular cartilage of knee joints in mice that develop OA as a result of a heterozygous mutation in type XI collagen. At the same time point, 6 months, we also found increased expression and activity of matrix metalloproteinase 13 (MMP-13) in the mutant mouse knee cartilage. The expression of both DDR2 and MMP-13 was increased in chondrocytes cultured on plates coated with native type II collagen but not on gelatin, and overexpression of DDR2, but not of a truncated form, was found to induce the expression of MMP-13 when chondrocytes were cultured on type II collagen but not on plastic. The DDR2-induced expression of MMP-13 appears to be specific, since we did not observe induction of MMP-1, MMP-3, MMP-8, ADAMTS-4, ADAMTS-5, and IL-1 transcripts in human chondrocytes or Mmp-3, Mmp-8, Adamts-4, Adamts-5, and Il-1 in mouse chondrocytes. Our data suggest that the defect in the cartilage matrix of mice that are heterozygous for a type XI collagen mutation (cho/+) permits activation and up-regulation of DDR2 in chondrocytes. This could be due to increased exposure of chondrocytes to type II collagen as a result of the decreased amount of type XI collagen in the mutant cartilage. The specific induction of MMP-13 by DDR2 in response to its cartilage-specific ligand, type II collagen, may contribute to cartilage damage in hereditary OA. Osteoarthritis (OA), 1The abbreviations used are: OA, osteoarthritis; MMP, metalloproteinase; DDR, discoidin domain receptor; PBS, phosphate-buffered saline; DMEM, Dulbecco's modified Eagle's medium. 1The abbreviations used are: OA, osteoarthritis; MMP, metalloproteinase; DDR, discoidin domain receptor; PBS, phosphate-buffered saline; DMEM, Dulbecco's modified Eagle's medium. the most common form of human arthritis (1Felson D.T. Lawrence R.C. Dieppe P.A. Hirsch R. Helmick C.G. Jordan J.M. Kington R.S. Lane N.E. Nevitt M.C. Zhang Y. Sowers M.F. McAlindon T. Spector T.D. Poole A.R. Yanovski S.Z. Ateshian G. Sharma L. Buckwalter J.A. Brandt K.D. Fries J.F. Ann. Intern. Med. 2000; 133: 635-646Crossref PubMed Scopus (1849) Google Scholar, 2Scott J.C. Lethbridge-Cejku M. Hochberg M.C. Reginster J.Y. Pelletier J.P. Martel-Pelletier J. Henrotin Y. Osteoarthritis: Clinical and Experimental Aspects. Springer, Milan, Italy1999: 20-52Google Scholar, 3Peyron J.G. Altman R.D. Moskowitz R.W. Howell D.S. Goldberg V.M. Mankin H.J. Ostoarthritis. W. B. Saunders Co., Philadelphia1992: 15-37Google Scholar), is considered a group of overlapping distinct diseases with different etiologies but with a similar clinical outcome. Although the causes of OA are diverse, mutations in cartilage specific collagen genes have been identified in human familial OA associated with a wide range of osteochondrodysplasia, from mild to lethal forms (4Spector T.D. MacGregor A.J. Osteoarthritis Cartilage. 2004; 12: S39-SS44Abstract Full Text Full Text PDF PubMed Scopus (361) Google Scholar, 5Simonet W.S. Mol. Genet. Metab. 2002; 77: 31-34Crossref PubMed Scopus (14) Google Scholar, 6Loughlin J. Rheum. Dis. Clin. North Am. 2002; 28: 95-109Abstract Full Text Full Text PDF PubMed Scopus (56) Google Scholar, 7Jimenez S.A. Dharmavaram R.M. Ann. Rheum. Dis. 1994; 53: 789-797Crossref PubMed Scopus (20) Google Scholar). In particular, mutations in type IX or XI collagens are associated with early-onset OA (8Diab M. Orthop. Rev. 1993; 22: 165-170PubMed Google Scholar, 9Reginato A.M. Olsen B.R. Arthritis Res. 2002; 4: 337-345Crossref PubMed Scopus (71) Google Scholar, 10Vikkula M. Olsen B.R. Ann. Med. 1996; 28: 301-304Crossref PubMed Scopus (17) Google Scholar). Mutations in genes encoding type IX collagen, COL9A1, COL9A2, and COL9A3, cause multiple epiphyseal dysplasia and intervertebral disc disease in humans. Multiple epiphyseal dysplasia is a milder dysplasia of the epiphyses of peripheral joints that is characterized by cartilage degeneration resulting in early-onset OA. One of the clinical features of intervertebral disc disease is the degeneration of the articular cartilage of the discs. Interestingly, the amount of type IX collagen is reduced in aged people (11Duance V.C. Wotton S.F. Biochem. Soc. Trans. 1991; 19: 376SCrossref PubMed Scopus (11) Google Scholar), in whom there is high incidence of OA. In humans, mutations in genes for type XI collagen, COL11A1 and COL11A2, have also been identified in several forms of familial OA (12Spranger J. Pediatr. Radiol. 1998; 28: 745-750Crossref PubMed Scopus (48) Google Scholar). A reduced amount of type XI collagen is also found in aged people (11Duance V.C. Wotton S.F. Biochem. Soc. Trans. 1991; 19: 376SCrossref PubMed Scopus (11) Google Scholar). The pathogenetic mechanisms underlying the OA due to these collagen mutations are largely unknown. We have used a mouse model of chondrodysplasia (cho) to investigate how mutations in type XI collagen cause an OA-like pathology. We found previously that the cho phenotype is due to a single nucleotide deletion leading to frameshift and premature termination of translation of the α1 chain of type XI collagen (13Li Y. Lacerda D.A. Warman M.L. Beier D.R. Yoshioka H. Ninomiya Y. Oxford J.T. Morris N.P. Andrikopoulos K. Ramirez F. Wardell B.B. Lifferth G.D. Teuscher C. Woodward S.R. Taylor B.A. Seegmiller R.E. Olsen B.R. Cell. 1995; 80: 423-430Abstract Full Text PDF PubMed Scopus (301) Google Scholar). Electron microscopy of the articular cartilage of knee joints in heterozygous mutant (cho/+) mice shows thick collagen fibrils (14Xu L. Flahiff C.M. Waldman B.A. Wu D. Olsen B.R. Setton L.A. Li Y. Arthritis Rheum. 2003; 48: 2509-2518Crossref PubMed Scopus (116) Google Scholar), which is one of the early clinical characteristics of human OA (15Weiss C. Fed. Proc. 1973; 32: 1459-1466PubMed Google Scholar). The cho/+ mice (homozygous cho/cho mice die at birth) reveal OA-like changes in their knee and temporomandibular joints, starting at the age of 3 months, without other apparent skeletal abnormalities (14Xu L. Flahiff C.M. Waldman B.A. Wu D. Olsen B.R. Setton L.A. Li Y. Arthritis Rheum. 2003; 48: 2509-2518Crossref PubMed Scopus (116) Google Scholar). We have also found increased protein expression of matrix metalloproteinase-3 (MMP-3, stromelysin) and MMP-13 (collagenase 3) in knee articular cartilage of cho/+ which is with a of human OA Arthritis Rheum. PubMed Scopus Google Scholar). Although a hereditary defect in type XI collagen is not for most forms of human OA, of matrix in cho/+ mice in the of and associated that human OA. of the cho/+ mouse is that the of with changes the of the a group of that are in of in most and as as and MMP-13 cartilage and to type II collagen and in articular cartilage Rev. PubMed Scopus Google Scholar, H. J.F. J. Full Text Full Text PDF PubMed Scopus Google Scholar). and are also in the cartilage matrix J.M. R. S.A. R. F. K. H. R. H. Y. H. K. M.C. R.C. J.M. PubMed Scopus Google Scholar, J. Biochem. PubMed Scopus Google Scholar). of these are at in articular cartilage. However, a of studies have that expression of these is increased in of human OA articular cartilage and that MMP-13 may be a in the of type II collagen in articular cartilage C. B. G. G. J. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, S.A. J. Clin. 1996; PubMed Scopus Google Scholar). This is with the OA-like changes in knee joints of mice that MMP-13 in cartilage L.A. L. W. L. J. J. Wu W. C. T. Poole A.R. J. Clin. PubMed Scopus Google Scholar). the of these to of cartilage is that their expression in chondrocytes is The discoidin domain and 2 were identified in and mice as cell surface receptor L. R. M. J. 1993; Full Text PDF PubMed Google Scholar, C. M. G. 1994; Google Scholar, T. B. G. H. K. 1993; Google Scholar, F. W. K. B. H. 1995; Google Scholar). The transcripts of the are in several human and mouse as skeletal and in that native collagens were the C. L. M. G. G.D. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, W. G.D. F. T. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). Interestingly, a of was not to of mice suggest that a role in and W. F. T. Mol. Cell. PubMed Scopus Google Scholar, G. W. J. Clin. PubMed Scopus Google Scholar). mice and a in the of J.P. J. C. K. J. G. G. C. R. PubMed Scopus Google Scholar). of DDR2 by type collagen the of in the human cell C. L. M. G. G.D. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). of DDR2 is associated with overexpression of in from arthritis J. H. Y. T. Li F. L. J. 2002; 19: PubMed Scopus Google Scholar). type XI collagen is an of the type II collagen fibrils in we that the cho mutation may result in a of the collagen in cartilage matrix and the chondrocytes and type II collagen We that the expression of a cell surface receptor with native collagen with other matrix as or cartilage matrix protein is increased in the articular cartilage of knee joints in cho/+ in indicate that up-regulation and activation of DDR2 by native type II collagen induce the expression of MMP-13 by chondrocytes. We also show that the increased expression of and is associated with type II collagen in cho/+ Our data suggest a and increased expression of receptor for of the collagen in articular cartilage leading to the and OA-like in cho/+ mouse of in of of cho/+ and cartilage were from the knee joints of cho/+ and mice at the age of 3 or 6 We used an to for metalloproteinase activity D.A. M.F. 2002; PubMed Scopus Google Scholar). The articular cartilage were from from cho/+ or mice at age and in a of the of and of were to of a for the a of of the and for a of different at and of the were was in was the with at and at activity was the of of in of cho/+ articular from cho/+ or mice at the of 3 or 6 were and the from the cho/+ or were for the of the The was with the were for by the of of the Mmp-3, and Mmp-8, and and Adamts-4, and Adamts-5, and and and different of were and The of the for Mmp-3, Mmp-8, Adamts-4, Adamts-5, and was of 3 at or of and of was the and the was at for 3 by of for for and for with a at for At the of the a a range and with was to the of the A in was in and was We used as The of by a was also by the amount of the amount of a and In when the of the is the of is considered In these the or of II in of cho/+ from cho/+ or mice at the of 3 and 6 were for The were in at by in for of were and with which the in type II collagen by The were with for the A without was also Electron has been in the (14Xu L. Flahiff C.M. Waldman B.A. Wu D. Olsen B.R. Setton L.A. Li Y. Arthritis Rheum. 2003; 48: 2509-2518Crossref PubMed Scopus (116) Google Scholar). knee joints from cho/+ mice and knee joints from their were The articular cartilage of the joints was and were in with at The were for from were and multiple in were for for were to the expression of and in the articular cartilage of knee joints from cho/+ and knee joints from different cho/+ mice and knee joints from different at the of 3 or 6 were The were in in at and in of were from the to the of were for The were for and with One of the was with an in at human human DDR2 human and human were from and used to The were with by with was with with a of and without was as a of in was as the for and at the of chondrocytes were from mice and cultured as in a L. L. Olsen B.R. Li Y. PubMed Scopus Google Scholar). The human cell of chondrocytes with a encoding in and R. J. J.F. J. Clin. 1994; PubMed Scopus Google were cultured in Dulbecco's modified Eagle's The were cultured in at and at a of at to 6 of type II collagen from cartilage was in at a of and used to We also plates coated with type II collagen and type II collagen as The type II collagen was at for and of the type II collagen was used to The other of the collagen was at to type II collagen and used for of A of native and type II was to to the of the plates coated with the collagen and plates were with mouse or human chondrocytes. The were cultured in for or from 3 for were and for and The was with were by the of of the for human for mouse DDR2, and MMP-1, and MMP-3, and MMP-8, and and ADAMTS-4, and ADAMTS-5, and and and The of the for the genes DDR2, MMP-1, MMP-3, MMP-8, ADAMTS-4, ADAMTS-5, and The was as of and of human DDR2, was from the human cell The was the The for the were to the human DDR2 The of the was with a and the of the was with an the was the expression The truncated of DDR2, the and was the which were with at the and The of the were by the or the truncated human DDR2 were and for chondrocytes the were in human were to in plates at in of 6 of and of were and for at in of was to and was for an at the was with of and the was to the cell in for at the was with an of and was for to expression of the and truncated DDR2 The were and to plates without or with with type II collagen and were for The of MMP-1, MMP-3, and MMP-13 were by and the were the same as was used as of with were cultured in type II plates for the to were as at at and 3) at protein was to were for and the were with and for in in of in cho/+ the articular cartilage that we previously in cho/+ mice was associated with increased of we metalloproteinase activity in of articular cartilage from knee a to we found that metalloproteinase activity was increased at the age of 3 in cho/+ mouse articular cartilage with cartilage from At the age of 6 months, metalloproteinase activity was increased in articular cartilage of cho/+ mice with indicate that the articular cartilage in cho/+ mice is associated with increased metalloproteinase of in of cho/+ which are in the articular cartilage of knee joints in cho/+ we the of Mmp-3, Mmp-8, Adamts-4, and We also since Il-1 may be associated with increased cartilage in of cartilage that the of was increased in cho/+ mice at the age of 6 mutant and mice was at 3 In the of the other were not increased or in articular cartilage from cho/+ mice with at the age of 6 suggest that may be a to the articular cartilage in cho/+ knee of II in of cho/+ the expression of in articular cartilage of cho/+ mice in the increased activity of the we the type II collagen in the articular cartilage by an that the of the by was in type II collagen at the age of 3 in cho/+ mice with mice not However, type II collagen were in cho/+ mice at the age of 6 We that type II collagen in the data suggest that the activity of is increased in cho/+ of and of a in of cho/+ type II collagen fibrils and a collagen receptor on chondrocytes may a role in in OA at in the of hereditary OA by type IX and type XI collagen In the articular cartilage of mouse knee joints, we type II collagen fibrils the but fibrils are in the In the increased of thick type II collagen fibrils were in the in cho/+ mice suggest the that the exposure of chondrocytes to type II collagen fibrils in the matrix is in cho/+ the expression of collagen on chondrocytes is by the cho/+ we the protein expression of different of collagen and and in the articular cartilage of knee joints at the of 3 and 6 in increased of were in the articular cartilage of cho/+ mice at the age of 6 with could be at 3 not was in the of when cho/+ and mice were at age and were by at both and could be cho/+ and type mice not the increased protein expression of was due to increased we the of in mice at the of 3 and 6 by in the of in cho/+ knee joints was not increased at the age of 3 months, but was increased with mice at the age of 6 data indicate that the expression of in the articular cartilage of cho/+ knee joints at 6 of age is increased at both the and protein DDR2 and MMP-13 in on II collagens are to DDR2 and induce expression of a metalloproteinase C. L. M. G. G.D. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, W. G.D. F. T. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar), we in to the of chondrocytes on type II plates on the expression of with characteristics of and matrix by were from cartilage. was in cultured mouse chondrocytes by a for chondrocytes Olsen B.R. 1993; PubMed Scopus Google Scholar), was that the in the were chondrocytes. The human cell was also used as a model to to We plates coated with native type II collagen and used plates as that the of MMP-13 was in human chondrocytes cultured on type II collagen for However, the of was not increased the The for MMP-3, MMP-8, ADAMTS-4, ADAMTS-5, and IL-1 were not we found that the of DDR2 in human chondrocytes was also increased on collagen for In mouse and were increased and in cultured in type II for with cultured in The of was not The for Mmp-8, Adamts-4, Adamts-5, and Il-1 were not in cultured mouse chondrocytes. indicate that chondrocytes to native type II collagen for are to increased of DDR2 and MMP-13 The of induction at that the response early events as activation of expression and protein We also the of the human chondrocytes to and type II collagen. on collagen on of expression of and DDR2 on type II collagen increased the of DDR2 and MMP-13 by and indicate that chondrocytes in a specific to type II collagen. MMP-13 in Human in to of DDR2 the DDR2 and we DDR2 human chondrocytes and cultured the on type II plates for in the of DDR2 was increased and MMP-13 was increased This result that overexpression of DDR2 and its activation by type II collagen in induction of MMP-13 in chondrocytes. that the MMP-13 response is to activation of DDR2, we a truncated DDR2 the human chondrocytes. the were cultured on type II plates for the of DDR2 and MMP-13 were reduced by with with In we also found that there was in the of MMP-13 in human chondrocytes with the or truncated DDR2 and cultured on plates for that activation of DDR2 MMP-13 and that the chondrocytes and type II collagen is for the of MMP-13 and DDR2 in chondrocytes. The the of MMP-13 in Human on II the in the up-regulation of MMP-13 by type II DDR2 we the of with protein and in the DDR2 in were but the of MMP-13 in chondrocytes cultured on type II plates with the was reduced to of the in cultured on type II plates in the of In chondrocytes with the other and the MMP-13 were the same as in the indicate that the not with the in up-regulation of DDR2 but of DDR2, resulting in decreased In we found that metalloproteinase activity was increased in articular cartilage of knee joints of cho/+ mice at the age of 3 and increased with This is with that the of articular cartilage degeneration in knee and temporomandibular joints of cho/+ mice is at the age of 3 we found that of several Adamts-4, and were not in the articular cartilage of knee joints in cho/+ mice at the age of 3 is that the increased activity at 3 is due to of protein time and to the and activation of the previously In we found that the protein expression of is increased in cho/+ mice at the age of 3 (14Xu L. Flahiff C.M. Waldman B.A. Wu D. Olsen B.R. Setton L.A. Li Y. Arthritis Rheum. 2003; 48: 2509-2518Crossref PubMed Scopus (116) Google Scholar). However, the increased protein expression is not in in cho/+ mice in by at a single time One is that the of the from cho/+ mice for may the of is that the expression and activity of in cho/+ mice at the age of 3 are we not observe the in (14Xu L. Flahiff C.M. Waldman B.A. Wu D. Olsen B.R. Setton L.A. Li Y. Arthritis Rheum. 2003; 48: 2509-2518Crossref PubMed Scopus (116) Google or in the of type II collagen and cho/+ mice at the age of 3 However, the in metalloproteinase activity with age in cho/+ mice at the age of 6 is associated with the in and protein In type II collagen in cho/+ articular as the of a high activity of of encoding other with and were not increased or were in the articular cartilage of knee joints of cho/+ suggest that and activity of an that as as collagen A.J. K. G. 1996; PubMed Scopus Google Scholar), may be a in the of OA in knee joints of cho/+ This is with that MMP-13 is in the articular cartilage of human OA joints Arthritis Rheum. PubMed Scopus Google Scholar, C. B. G. G. J. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, S.A. J. Clin. 1996; PubMed Scopus Google Scholar). a result of the type XI collagen mutation in cho thick type II collagen are and the of the type II collagen is (14Xu L. Flahiff C.M. Waldman B.A. Wu D. Olsen B.R. Setton L.A. Li Y. Arthritis Rheum. 2003; 48: 2509-2518Crossref PubMed Scopus (116) Google Scholar). Although the amount of type II collagen in the articular cartilage of mutant knee joints may not be the of type II collagen in the articular cartilage is the increased of thick type II collagen in with chondrocytes. This of type II collagen may the chondrocytes and type II collagen resulting in the expression of a collagen Our data that the protein and of are increased in the articular cartilage of cho/+ mice at the age of 6 We did not an expression of other collagen and on chondrocytes. This that the up-regulation of and protein may result from the receptor to its ligand, type II collagen, in cho/+ The the increased of and protein in the articular cartilage of cho/+ mice to investigate a role of in the expression and of other in chondrocytes. from in that the of MMP-13 is increased in human and mouse chondrocytes when the are cultured on type II plates for and that the of other MMP-1, MMP-3, MMP-8, ADAMTS-4, and are not increased or Our data also that the of DDR2 is increased in cultured on type II collagen. we that of a DDR2 in human chondrocytes in of MMP-13 In the of MMP-13 is reduced of a truncated DDR2, the human chondrocytes cultured on type II collagen. the of these we that DDR2 in up-regulation of MMP-13 in chondrocytes cultured on type II In we that chondrocytes and type II collagen is since the MMP-13 is not in when are cultured on with or truncated DDR2 The data indicate that the activation of DDR2 by type II collagen, not expression of but also the expression of DDR2 in chondrocytes. there is on the by DDR2, we to these are in the up-regulation of MMP-13 by DDR2 on we that the is in the increased expression of MMP-13 by DDR2 activation but not in the up-regulation of DDR2 has been that IL-1 induce MMP-13 expression in chondrocytes and that by the and are for of MMP-13 J.A. Res. PubMed Scopus Google Scholar, J.A. Arthritis Rheum. 2000; PubMed Scopus Google Scholar). IL-1 is an in the of MMP-13 expression by activation of DDR2, we IL-1 in mouse and human chondrocytes cultured on type II collagen for or IL-1 was in the chondrocytes cultured on type II is that IL-1 a role in the of MMP-13 expression by DDR2 activation in The activation of native collagens as type and collagens C. L. M. G. G.D. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, W. G.D. F. T. Mol. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). Interestingly, we found that the increased of thick type II collagen in in cho/+ articular cartilage. We that the and of type II collagen fibrils in the of cho/+ mice may the chondrocytes and type II collagen a DDR2 is to a of events leading to and activation of as MMP-13 resulting in OA. The was a by Poole at for

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.