Gene transfer of tissue inhibitor of metalloproteinases-3 reverses the inhibitory effects of TNF-alpha on Fas-induced apoptosis in rheumatoid arthritis synovial fibroblasts.
Apart from counteracting matrix metalloproteinases, tissue inhibitor of metalloproteinases-3 (TIMP-3) has proapoptotic properties. These features have been attributed to the inhibition of metalloproteinases involved in the shedding of cell surface receptors such as the TNFR. However, little is known about effects of TIMP-3 in cells that are not susceptible to apoptosis by TNF-alpha. In this study, we report that gene transfer of TIMP-3 into human rheumatoid arthritis synovial fibroblasts and MRC-5 human fetal lung fibroblasts facilitates apoptosis and completely reverses the apoptosis-inhibiting effects of TNF-alpha. Although TNF-alpha inhibits Fas/CD95-induced apoptosis in untransfected and mock-transfected cells, fibroblasts ectopically expressing TIMP-3 are sensitized most strongly to Fas/CD95-mediated cell death by TNF-alpha. Neither synthetic MMP inhibitors nor glycosylated bioactive TIMP-3 are able to achieve these effects. Gene transfer of TIMP-3 inhibits the TNF-alpha-induced activation of NF-kappaB in rheumatoid arthritis synovial fibroblasts and reduces the up-regulation of soluble Fas/CD95 by TNF-alpha, but has no effects on the cell surface expression of Fas. Collectively, our data demonstrate that intracellularly produced TIMP-3 not only induces apoptosis, but also modulates the apoptosis-inhibiting effects of TNF-alpha in human rheumatoid arthritis synovial fibroblast-like cells. Thus, our findings may stimulate further studies on the therapeutic potential of gene transfer strategies with TIMP-3.
Drynda
Andreas
A
Division of Experimental Rheumatology and Orthopedics, Institute of Microbiology, Otto-von-Guericke University, Magdeburg, Germany.
Quax
Paul H A
PH
Neumann
Manfred
M
van der Laan
Willemijn H
WH
Pap
Géza
G
Drynda
Susanne
S
Meinecke
Ingmar
I
Kekow
Joern
J
Neumann
Wolfram
W
Huizinga
Tom W J
TW
Naumann
Michael
M
König
Wolfgang
W
Pap
Thomas
T
D000260:Adenoviruses, Human / Q000235:genetics; D017209:Apoptosis / Q000235:genetics / Q000276:immunology*; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D002460:Cell Line; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D005347:Fibroblasts / Q000276:immunology* / Q000473:pathology; D018014:Gene Transfer Techniques; D006801:Humans; D007424:Intracellular Fluid / Q000201:enzymology / Q000276:immunology; D008297:Male; D008875:Middle Aged; D018716:Molecular Mimicry / Q000187:drug effects / Q000276:immunology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D011994:Recombinant Proteins / Q000138:chemical synthesis / Q000494:pharmacology; D012995:Solubility; D013583:Synovial Membrane / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D019717:Tissue Inhibitor of Metalloproteinase-3; D019714:Tissue Inhibitor of Metalloproteinases / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000502:physiology*; D019014:fas Receptor / Q000096:biosynthesis / Q000502:physiology*
D016328:NF-kappa B; D011994:Recombinant Proteins; C088390:TIMP3 protein, human; D019717:Tissue Inhibitor of Metalloproteinase-3; D019714:Tissue Inhibitor of Metalloproteinases; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor