Stimulation of collagenase 3 expression in synovial fibroblasts of patients with rheumatoid arthritis by contact with a three-dimensional collagen matrix or with normal cartilage when coimplanted in NOD/SCID mice.
OBJECTIVE
To study the expression of collagenase 3 (matrix metalloproteinase 13 [MMP-13]) and collagenase 1 (MMP-1) in synovial fibroblasts from patients with rheumatoid arthritis (RA) when cultured within 3-dimensional collagen gels or coimplanted with normal cartilage in immunodeficient NOD/SCID mice.


METHODS
Messenger RNA (mRNA) and protein expression of collagenase 3 and collagenase 1 were characterized in synovial and skin fibroblasts by Northern blot and Western blot analysis. The mRNA expression of both collagenases in cell-cartilage implants in NOD/SCID mice was investigated by in situ hybridization in combination with immunohistochemistry of human fibroblasts.


RESULTS
Synovial fibroblasts coimplanted with normal cartilage in NOD/SCID mice deeply invaded adjacent cartilage tissue. In this in vivo system of cartilage destruction, collagenase 3 mRNA was induced in synovial fibroblasts at sites of cartilage erosion, while the expression of collagenase 1 mRNA could not be detected. Culture of synovial fibroblasts within 3-dimensional collagen gels was associated with a marked increase in collagenase 3 mRNA expression and proenzyme production. This stimulatory effect was 1 order of magnitude higher in comparison with a 2-4-fold increase upon treatment with interleukin-1beta or tumor necrosis factor a. In contrast, mRNA expression and proenzyme production of collagenase 1 were increased strongly, and to a similar extent, either by contact with 3-dimensional collagen or by proinflammatory cytokines.


CONCLUSIONS
The expression of collagenase 3, in contrast to that of collagenase 1, is preferentially stimulated in synovial fibroblasts by 3-dimensional collagen rather than by proinflammatory cytokines. The induction of collagenase 3 by cell-matrix interactions represents a potential mechanism contributing to the invasive phenotype of synovial fibroblasts at sites of synovial invasion into cartilage in RA.
Wernicke
Dirk
D
Max-Delbrück-Center for Molecular Medicine, and Clinic of Rheumatology, Berlin-Buch, Germany. dwernic@mdc-berlin.de
Schulze-Westhoff
Claudia
C
Bräuer
Rolf
R
Petrow
Peter
P
Zacher
Josef
J
Gay
Steffen
S
Gromnica-Ihle
Erika
E
D000818:Animals; D001172:Arthritis, Rheumatoid / Q000201:enzymology / Q000503:physiopathology*; D002356:Cartilage / Q000166:cytology* / Q000637:transplantation; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D017364:Collagenases / Q000032:analysis / Q000235:genetics*; D005347:Fibroblasts / Q000201:enzymology / Q000637:transplantation; D005782:Gels; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D016867:Immunocompromised Host; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D053509:Matrix Metalloproteinase 13; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D012333:RNA, Messenger / Q000032:analysis; D013583:Synovial Membrane / Q000166:cytology* / Q000637:transplantation
D005782:Gels; D012333:RNA, Messenger; D003094:Collagen; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; C507081:Mmp13 protein, mouse; C417249:collagenase 1