Research Article
Proline-rich tyrosine kinase 2 and Src kinase signaling transduce monosodium urate crystal–induced nitric oxide production and matrix metalloproteinase 3 expression in chondrocytes
Ru Liu,
Frédéric Lioté,
David M. Rose,
Denise Merz,
Robert Terkeltaub,
Ru Liu
VA Medical Center and University of California, San Diego
Drs. Liu and Lioté contributed equally to this work.
Search for more papers by this authorFrédéric Lioté
Hôpital Lariboisière, Paris 7 University, Paris, France
Drs. Liu and Lioté contributed equally to this work.
Search for more papers by this authorDavid M. Rose
VA Medical Center and University of California, San Diego
Search for more papers by this authorDenise Merz
VA Medical Center and University of California, San Diego
Search for more papers by this authorCorresponding Author
Robert Terkeltaub
VA Medical Center and University of California, San Diego
VA Medical Center, MC111K, 3350 La Jolla Village Drive, San Diego, CA 92161Search for more papers by this authorRu Liu,
Frédéric Lioté,
David M. Rose,
Denise Merz,
Robert Terkeltaub,
Ru Liu
VA Medical Center and University of California, San Diego
Drs. Liu and Lioté contributed equally to this work.
Search for more papers by this authorFrédéric Lioté
Hôpital Lariboisière, Paris 7 University, Paris, France
Drs. Liu and Lioté contributed equally to this work.
Search for more papers by this authorDavid M. Rose
VA Medical Center and University of California, San Diego
Search for more papers by this authorDenise Merz
VA Medical Center and University of California, San Diego
Search for more papers by this authorCorresponding Author
Robert Terkeltaub
VA Medical Center and University of California, San Diego
VA Medical Center, MC111K, 3350 La Jolla Village Drive, San Diego, CA 92161Search for more papers by this authorAbstract
Objective
Articular deposition of monosodium urate monohydrate (MSU) crystals may promote cartilage and bone erosion. Therefore, the aim of this study was to determine how MSU crystals stimulate chondrocytes.
Methods
Nitric oxide (NO) release, and expression of inducible nitric oxide synthase (iNOS) and matrix metalloproteinase 3 (MMP-3) were assessed in cultured chondrocytes treated with MSU. MSU-induced functional signaling by specific protein kinases (p38, Src, and the focal adhesion kinase [FAK] family members proline-rich tyrosine kinase 2 [Pyk-2] and FAK) was also examined using selective pharmacologic inhibitors and transfection of kinase mutants.
Results
MSU induced MMP-3 and iNOS expression and NO release in chondrocytes in a p38-dependent manner that did not require interleukin-1 (IL-1), as demonstrated by using IL-1 receptor antagonist. MSU induced rapid tyrosine phosphorylation of Pyk-2 and FAK, their adaptor protein paxillin, and interacting kinase c-Src. Pyk-2 and c-Src signaling both mediated p38 MAPK activation in response to MSU. Pyk-2 and c-Src signaling played a major role in transducing MSU-induced NO production and MMP-3 expression. But, despite the observed FAK phosphorylation, a selective pharmacologic FAK inhibitor and a FAK dominant-negative mutant both failed to block MSU-induced NO release or MMP-3 expression in parallel experiments.
Conclusion
In chondrocytes, MSU crystals activate a signaling kinase cascade typically employed by adhesion receptors that involves upstream Src and FAK family activation and downstream p38 activation. In this cascade, Pyk-2, Src, and p38 kinases transduce MSU-induced NO production and MMP-3 expression. Our results identify Pyk-2 and c-Src as novel sites for potential therapeutic intervention in cartilage degradation in chronic gout.
REFERENCES
- 1 Dieppe P, Bacon P, Bamji A, Watt I. Gout. In: JH Klippel and P Dieppe, editors. Slide Atlas of Rheumatology. London: Gower Medical Publishing; 1985.
- 2 Hasselbacher P. Stimulation of synovial fibroblasts by calcium oxalate and monosodium urate monohydrate: a mechanism of connective tissue degradation in oxalosis and gout. J Lab Clin Med 1982; 100: 977–85.
- 3 Schiltz C, Liote F, Prudhommeaux F, Meunier A, Champy R, Callebert J, et al. Monosodium urate monohydrate crystal-induced inflammation in vivo: quantitative histomorphometric analysis of cellular events. Arthritis Rheum 2002; 46: 1643–50.
- 4 Yagnik DR, Hillyer P, Marshall D, Smythe CD, Krausz T, Haskard DO, et al. Noninflammatory phagocytosis of monosodium urate monohydrate crystals by mouse macrophages: implications for the control of joint inflammation in gout. Arthritis Rheum 2000; 43: 1779–89.
- 5 Wigley FM, Fine IT, Newcombe DS. The role of the human synovial fibroblast in monosodium urate crystal-induced synovitis. J Rheumatol 1983; 10: 602–11.
- 6 Pouliot M, James MJ, McColl SR, Naccache PH, Cleland LG. Monosodium urate microcrystals induce cyclooxygenase-2 in human monocytes. Blood 1998; 91: 1769–76.
- 7 Di Giovine FS, Malawista SE, Thornton E, Duff GW. Urate crystals stimulate production of tumor necrosis factor α from human blood monocytes and synovial cells: cytokine mRNA and protein kinetics, and cellular distribution. J Clin Invest 1991; 87: 1375–81.
- 8 Di Giovine FS, Malawista SE, Nuki G, Duff, GW. Interleukin 1 (IL 1) as a mediator of crystal arthritis: stimulation of T cell and synovial fibroblast mitogenesis by urate crystal-induced IL 1. J Immunol 1987; 138: 3213–8.
- 9 Guerne PA, Terkeltaub R, Zuraw B, Lotz M. Inflammatory microcrystals stimulate interleukin-6 production and secretion by human monocytes and synoviocytes. Arthritis Rheum 1989; 32: 1443–52.
- 10 Terkeltaub R, Zachariae C, Santoro D, Martin J, Peveri P, Matsushima K. Monocyte-derived neutrophil chemotactic factor/interleukin-8 is a potential mediator of crystal-induced inflammation. Arthritis Rheum 1991; 34: 894–903.
- 11 Barabe F, Gilbert C, Liao N, Bourgoin SG, Naccache PH. Crystal-induced neutrophil activation. VI. Involvement of FcγRIIIB (CD16) and CD11b in response to inflammatory microcrystals. FASEB J 1998; 12: 209–20.
- 12 Desaulniers P, Fernandes M, Gilbert C, Bourgoin SG, Naccache PH. Crystal-induced neutrophil activation. VII. Involvement of Syk in the responses to monosodium urate crystals. J Leukoc Biol 2001; 70: 659–68.
- 13 Liu R, Aupperle K, Terkeltaub R. Src family protein tyrosine kinase signaling mediates monosodium urate crystal-induced IL-8 expression by monocytic THP-1 cells. J Leukoc Biol 2001; 70: 961–8.
- 14 Liu R, O'Connell M, Johnson K, Pritzker K, Mackman N, Terkeltaub R. Extracellular signal-regulated kinase 1/extracellular signal-regulated kinase 2 mitogen-activated protein kinase signaling and activation of activator protein 1 and nuclear factor κB transcription factors play central roles in interleukin-8 expression stimulated by monosodium urate monohydrate and calcium pyrophosphate crystals in monocytic cells. Arthritis Rheum 2000; 43: 1145–55.
- 15 McCarty DJ, Cheung HS. Prostaglandin (PG) E2 generation by cultured canine synovial fibroblasts exposed to microcrystals containing calcium. Ann Rheum Dis 1985; 44: 316–20.
- 16 McMillan RM, Vater CA, Hasselbacher P, Hahn J, Harris ED Jr. Induction of collagenase and prostaglandin synthesis in synovial fibroblasts treated with monosodium urate crystals. J Pharm Pharmacol 1981; 33: 382–3.
- 17 Hasselbacher P, McMillan RM, Vater CA, Hahn J, Harris ED Jr. Stimulation of secretion of collagenase and prostaglandin E2 by synovial fibroblasts in response to crystals of monosodium urate monohydrate: a model for joint destruction in gout. Trans Assoc Am Physicians 1981; 94: 243–52.
- 18 Bouchard L, de Medicis R, Lussier A, Naccache PH, Poubelle PE. Inflammatory microcrystals alter the functional phenotype of human osteoblast-like cells in vitro: synergism with IL-1 to overexpress cyclooxygenase-2. J Immunol 2002; 168: 5310–7.
- 19 Goldring MB. 2000. The role of the chondrocyte in osteoarthritis. Arthritis Rheum 2000; 43: 1916–26.
- 20 Palmer RMJ, Hickery MS, Charles IG, Moncada S, Bayliss MT. Induction of nitric oxide synthase in human chondrocytes. Biochem Biophys Res Commun 1993; 193: 398–405.
- 21 Maier R, Bilbe G, Rediske J, Lotz M. Inducible nitric oxide synthase from human articular chondrocytes: cDNA cloning and analysis of mRNA expression. Biochim Biophys Acta 1994; 1208: 145–50.
- 22 Blanco FJ, Lotz M. IL-1-induced nitric oxide inhibits chondrocyte proliferation via PGE2. Exp Cell Res 1995; 218: 319–25.
- 23 Taskiran D, Stefanovic-Racic M, Georgescu H, Evans C. Nitric oxide mediates suppression of cartilage proteoglycan synthesis by interleukin-1. Biochem Biophys Res Commun 1994; 200: 142–8.
- 24 Blanco FJ, Ochs RL, Schwarz H, Lotz M. Chondrocyte apoptosis induced by nitric oxide. Am J Pathol 1995; 146: 75–85.
- 25 Murrell GA, Jang D, Williams RJ. Nitric oxide activates metalloprotease enzymes in articular cartilage. Biochem Biophys Res Commun 1995; 206: 15–21.
- 26 Gu Z, Kaul M, Yan B, Kridel SJ, Cui J, Strongin A, et al. S-nitosylation of matrix metalloproteinases: signaling pathway to neuronal cell death. Science 2002; 297: 1186–90.
- 27 Jaques BC, Ginsberg MH. The role of cell surface proteins in platelet stimulation by monosodium urate crystals. Arthritis Rheum 1982; 25: 508–21.
- 28 Loeser RF. Integrins and cell signaling in chondrocytes. Biorheology 2002; 39: 119–24.
- 29 Schlaepfer DD, Hauck CR, Sieg DJ. Signaling through focal adhesion kinase. Prog Biophys Mol Biol 1999; 71: 435–78.
- 30 Terkeltaub R, Zachariae C, Santoro D, Martin J, Peveri P, Matsushima K. Monocyte-derived neutrophil chemotactic factor/interleukin-8 is a potential mediator of crystal-induced inflammation. Arthritis Rheum 1991; 34: 894–903.
- 31 Reginato AM, Iozzo RV, Jimenez SA. Formation of nodular structures resembling mature articular cartilage in long-term primary cultures of human fetal epiphyseal chondrocytes on a hydrogel substrate. Arthritis Rheum 1994; 37: 1338–49.
- 32 Johnson K, Farley D, Hu SI, Terkeltaub R. One of two chondrocyte-expressed isoforms of cartilage intermediate-layer protein functions as an insulin-like growth factor 1 antagonist. Arthritis Rheum 2003; 48: 1302–14.
- 33 Johnson K, Vaingankar S, Chen Y, Moffa A, Goldring MB, Sano K, et al. Differential mechanisms of inorganic pyrophosphate production by plasma cell membrane glycoprotein-1 and B10 in chondrocytes. Arthritis Rheum 1999; 42: 1986–97.
- 34 Hashimoto S, Takahashi K, Ochs RL, Coutts RD, Amiel D, Lotz M. Nitric oxide production and apoptosis in cells of the meniscus during experimental osteoarthritis. Arthritis Rheum 1999; 42: 2123–31.
- 35 Huang R, Rosenbach M, Vaughn R, Provvedini D, Rebbe N, Hickman S, et al. Expression of the murine plasma cell nucleotide pyrophosphohydrolase PC-1 is shared by human liver, bone, and cartilage cells: regulation of PC-1 expression in osteosarcoma cells by transforming growth factor-β. J Clin Invest 1994; 94: 560–7.
- 36 Goomer RS, Johnson KA, Burton DW, Amiel D, Maris TM, Gujral A, et al. The tetrabasic KKKK147–150 motif determines intracrine regulatory effects of PTHrP 1–173 on chondrocyte PPi metabolism and matrix synthesis. Endocrinology 2000; 141: 4613–22.
- 37 Akahoshi T, Nagaoka T, Namai R, Sekiyama N, Kondo H. Prevention of neutrophil apoptosis by monosodium urate crystals. Rheumatol Int 1997; 16: 231–5.
- 38 Stadler J, Stefanovic-Racic M, Billiar TR, Curran RD, McIntyre LA, Georgescu HI, et al. Articular chondrocytes synthesize nitric oxide in response to cytokines and lipopolysaccharide. J Immunol 1991; 147: 3915–20.
- 39 Goldring MB, Birkhead JR, Suen LF, Yamin R, Mizuno S, Glowacki J, et al. Interleukin-1β-modulated gene expression in immortalized human chondrocytes. J Clin Invest 1994; 94: 2307–16.
- 40 Attur MG, Dave M, Cipolletta C, Kang P, Goldring MB, Patel IR, et al. Reversal of autocrine and paracrine effects of interleukin 1 (IL-1) in human arthritis by type II IL-1 decoy receptor: potential for pharmacological intervention. J Biol Chem 2000; 275: 40307–15.
- 41 Fuortes M, Melchior M, Han H, Lyon GJ, Nathan C. Role of the tyrosine kinase pyk2 in the integrin-dependent activation of human neutrophils by TNF. J Clin Invest 1999; 104: 327–35.
- 42 Tsuda T, Kusui T, Jensen RT. Neuromedin B receptor activation causes tyrosine phosphorylation of p125FAK by a phospholipase C independent mechanism which requires p21ρ and integrity of the actin cytoskeleton. Biochemistry 1997; 36: 16328–37.
- 43 Schaller MD, Borgman CA, Parsons JT. Autonomous expression of a noncatalytic domain of the focal adhesion-associated protein tyrosine kinase pp125FAK. Mol Cell Biol 1993; 13: 785–91.
- 44 Dikic I, Tokiwa G, Lev S, Courtneidge SA, Schelessinger J. A role for Pyk2 and Src in linking G-protein-coupled receptors with MAP kinase activation. Nature 1996; 383: 547–50.
- 45 Okada M, Nada S, Yamanashi Y, Yamamoto T, Nakagawa H. CSK: a protein-tyrosine kinase involved in regulation of src family kinases. J Biol Chem 1991; 266: 24249–52.
- 46 Tokiwa G, Dikic I, Lev S, Schlessinger J. Activation of Pyk2 by stress signals and coupling with JNK signaling pathway. Science 1996; 273: 792–4.
- 47 Pandey P, Avraham S, Kumar S, Nakazawa A, Place A, Ghanem L, et al. Activation of p38 mitogen-activated protein kinase by PYK2/related adhesion focal tyrosine kinase-dependent mechanism. J Biol Chem 1999; 274: 10140–4.
- 48 Liote F. Hyperuricemia and gout. Curr Rheumatol Rep 2003; 5: 227–34.
- 49 McCarthy GM, Barthelemy CR, Veum JA, Wortmann RL. Influence of antihyperuricemic therapy on the clinical and radiographic progression of gout. Arthritis Rheum 1991; 34: 1489–94.
- 50 Abramson SB, Attur M, Amin AR, Clancy R. Nitric oxide and inflammatory mediators in the perpetuation of osteoarthritis. Curr Rheumatol Rep 2001; 3: 535–41.
- 51 Smith RL. Degradative enzymes in osteoarthritis. Front Biosci 1999; 4: D704–12.
- 52 Mendes AF, Caramona MM, Carvalho AP, Lopes MC. Role of mitogen-activated protein kinases and tyrosine kinases on IL-1-induced NF-κB activation and iNOS expression in bovine articular chondrocytes. Nitric Oxide 2002; 6: 35–44.
- 53 Reuben PM, Brogley MA, Sun Y, Cheung HS. Molecular mechanism of the induction of metalloproteinases 1 and 3 in human fibroblasts by basic calcium phosphate crystals: role of calcium-dependent protein kinase Cα. J Biol Chem 2002; 277: 15190–8.
- 54 Avraham H, Park SY, Schinkmann K, Avraham S. RAFTK/Pyk2-mediated cellular signalling. Cell Signal 2000; 12: 123–33.
- 55 Schaller MD. Paxillin, a focal adhesion-associated adaptor protein. Oncogene 2001; 20: 6459–72.
- 56 Klingbeil CK, Hauck CR, Hsia DA, Jones KC, Reider SR, Schlaepfer DD. Targeting Pyk2 to β1-integrin–containing focal contacts rescues fibronectin-stimulated signaling and haptotactic motility defects of focal adhesion kinase–null cells. J Cell Biol 2001; 152: 97–110.
- 57 Dunty JM, Schaller MD. The N termini of focal adhesion kinase family members regulate substrate phosphorylation, localization, and cell morphology. J Biol Chem 2002; 277: 45644–54.
- 58 Burt HM, Jackson JK. Cytosolic Ca2+ concentration determinations in neutrophils stimulated by monosodium urate and calcium pyrophosphate crystals: effect of protein adsorption. J Rheumatol 1994; 21: 138–44.
- 59 Woods VL Jr, Schreck PJ, Gesink DS, Pacheco HO, Amiel D, Akeson WH, et al. Integrin expression by human articular chondrocytes. Arthritis Rheum 1994; 37: 537–44.
- 60 Loeser RF, Carlson CS, McGee MP. Expression of β1 integrins by cultured articular chondrocytes and in osteoarthritic cartilage. Exp Cell Res 1995; 217: 248–57.
- 61 Attur MG, Dave MN, Clancy RM, Patel IR, Abramson SB, Amin AR. Functional genomic analysis in arthritis-affected cartilage: yin-yang regulation of inflammatory mediators by α5β1 and βVβ3 integrins. J Immunol 2000; 164: 2684–91.
- 62 Forsyth CB, Pulai J, Loeser RF. Fibronectin fragments and blocking antibodies to α2β1 and α5β1 integrins stimulate mitogen-activated protein kinase signaling and increase collagenase 3 (matrix metalloproteinase 13) production by human articular chondrocytes. Arthritis Rheum 2002; 46: 2368–76.
- 63 Loeser RF Jr, Forsyth C, Im H-J. Fibronectin-fragment activation of Pyk-2 mediates integrin signals regulating collgenase-3 expression by chondrocytes. Arthritis Rheum 2002; 46(Suppl): S372.
