NF-κB Mediates the Stimulation of Cytokine and Chemokine Expression by Human Articular Chondrocytes in Response to Fibronectin Fragments1

Fibronectin fragments (FN-f) that bind to the α5β1 integrin stimulate chondrocyte-mediated cartilage destruction and could play an important role in the progression of arthritis. The objective of this study was to identify potential cytokine mediators of cartilage inflammation and destruction induced by FN-f and to investigate the mechanism of their stimulation. Human articular chondrocytes, isolated from normal ankle cartilage obtained from tissue donors, were treated with a 110-kDa FN-f in serum-free culture, and expression of various cytokine genes was analyzed by cDNA microarray and by a cytokine protein array. Compared with untreated control cultures, stimulation by FN-f resulted in a >2-fold increase in IL-6, IL-8, MCP-1, and growth-related oncogene β (GRO-β). Constitutive and FN-f-inducible expression of GRO-α and GRO-γ were also noted by RT-PCR and confirmed by immunoblotting. Previous reports of IL-1β expression induced by FN-f were also confirmed, while TNF expression was found to be very low. Inhibitor studies revealed that FN-f-induced stimulation of chondrocyte chemokine expression was dependent on NF-κB activity, but independent of IL-1 autocrine signaling. The ability of FN-f to stimulate chondrocyte expression of multiple proinflammatory cytokines and chemokines suggests that damage to the cartilage matrix is capable of inducing a proinflammatory state responsible for further progressive matrix destruction, which also includes the chemoattraction of inflammatory cells. Targeting the signaling pathways activated by FN-f may be an effective means of inhibiting production of multiple mediators of cartilage destruction.

[1]  M. Feldmann,et al.  Heterogeneous requirement of IkappaB kinase 2 for inflammatory cytokine and matrix metalloproteinase production in rheumatoid arthritis: implications for therapy. , 2003, Arthritis and rheumatism.

[2]  T. Hughes,et al.  Signaling and circuitry of multiple MAPK pathways revealed by a matrix of global gene expression profiles. , 2000, Science.

[3]  Z. Werb,et al.  Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression , 1989, The Journal of cell biology.

[4]  A. Samarel,et al.  Fibronectin Fragment Activation of Proline-rich Tyrosine Kinase PYK2 Mediates Integrin Signals Regulating Collagenase-3 Expression by Human Chondrocytes through a Protein Kinase C-dependent Pathway* , 2003, Journal of Biological Chemistry.

[5]  R. Terkeltaub,et al.  IL-8/CXCL8 and Growth-Related Oncogene α/CXCL1 Induce Chondrocyte Hypertrophic Differentiation 1 , 2003, The Journal of Immunology.

[6]  M. Lotz,et al.  Mechanisms of chondrocyte apoptosis. , 1999, Osteoarthritis and cartilage.

[7]  S. Abramson,et al.  Functional Genomic Analysis in Arthritis-Affected Cartilage: Yin-Yang Regulation of Inflammatory Mediators by α5β1 and αVβ3 Integrins1 , 2000, The Journal of Immunology.

[8]  A. Facchini,et al.  Chemokine production by human chondrocytes. , 1999, The Journal of rheumatology.

[9]  P. Guerne,et al.  IL-6 production by human articular chondrocytes. Modulation of its synthesis by cytokines, growth factors, and hormones in vitro. , 1990, Journal of immunology.

[10]  P. Kapahi,et al.  Hypoestoxide, a novel anti-inflammatory natural diterpene, inhibits the activity of IkappaB kinase. , 2001, Cellular immunology.

[11]  R. Borzì,et al.  Down-modulation of chemokine receptor cartilage expression in inflammatory arthritis. , 2003, Rheumatology.

[12]  R. Borzì,et al.  Flow cytometric analysis of intracellular chemokines in chondrocytes in vivo: constitutive expression and enhancement in osteoarthritis and rheumatoid arthritis , 1999, FEBS letters.

[13]  T. Aigner,et al.  Subtyping of osteoarthritic synoviopathy. , 2002, Clinical and experimental rheumatology.

[14]  M. Baggiolini,et al.  Human chondrocytes express functional chemokine receptors and release matrix-degrading enzymes in response to C-X-C and C-C chemokines. , 2000, Arthritis and rheumatism.

[15]  E. Ruoslahti,et al.  Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor , 1985, Cell.

[16]  R. Loeser,et al.  The α5β1 integrin provides matrix survival signals for normal and osteoarthritic human articular chondrocytes in vitro , 2002 .

[17]  M. Karin,et al.  c-Jun N-terminal kinase is required for metalloproteinase expression and joint destruction in inflammatory arthritis. , 2001, The Journal of clinical investigation.

[18]  D. Xie,et al.  Fibronectin fragments alter matrix protein synthesis in cartilage tissue cultured in vitro. , 1993, Archives of biochemistry and biophysics.

[19]  G. Homandberg,et al.  Cartilage chondrolysis by fibronectin fragments is associated with release of several proteinases: stromelysin plays a major role in chondrolysis. , 1994, Archives of biochemistry and biophysics.

[20]  Qian Chen,et al.  Stimulation of matrix metalloprotease 3 release from human chondrocytes by the interaction of stromal cell-derived factor 1 and CXC chemokine receptor 4. , 2002, Arthritis and rheumatism.

[21]  R. Gatti,et al.  Growth‐related oncogene α induction of apoptosis in osteoarthritis chondrocytes , 2002 .

[22]  J. Boehm,et al.  Disease-modifying activity of SB 242235, a selective inhibitor of p38 mitogen-activated protein kinase, in rat adjuvant-induced arthritis. , 2000, Arthritis and rheumatism.

[23]  M. Goldring,et al.  The role of the chondrocyte in osteoarthritis. , 2000, Arthritis and rheumatism.

[24]  D. Zukor,et al.  Sites of collagenase cleavage and denaturation of type II collagen in aging and osteoarthritic articular cartilage and their relationship to the distribution of matrix metalloproteinase 1 and matrix metalloproteinase 13. , 2002, Arthritis and rheumatism.

[25]  A. Recklies,et al.  Induction of synthesis and release of interleukin-8 from human articular chondrocytes and cartilage explants. , 1992, Arthritis and rheumatism.

[26]  R. Mason,et al.  Gene deletion of either interleukin-1beta, interleukin-1beta-converting enzyme, inducible nitric oxide synthase, or stromelysin 1 accelerates the development of knee osteoarthritis in mice after surgical transection of the medial collateral ligament and partial medial meniscectomy. , 2003, Arthritis and rheumatism.

[27]  M. Kagnoff,et al.  Differential and regulated expression of C-X-C, C-C, and C-chemokines by human colon epithelial cells. , 1997, Gastroenterology.

[28]  L. Zardi,et al.  Characterization of synovial fluid fibronectin from patients with rheumatic inflammatory diseases and healthy subjects. , 1984, Arthritis and rheumatism.

[29]  G. Homandberg,et al.  Fibronectin-fragment-induced cartilage chondrolysis is associated with release of catabolic cytokines. , 1997, The Biochemical journal.

[30]  G. Homandberg,et al.  Fibronectin fragments in osteoarthritic synovial fluid. , 1992, The Journal of rheumatology.

[31]  M. Tortorella,et al.  Signal transduction through chondrocyte integrin receptors induces matrix metalloproteinase synthesis and synergizes with interleukin-1. , 1995, Arthritis and rheumatism.

[32]  A. V. van Wijnen,et al.  Inhibitory Effects of Insulin-like Growth Factor-1 and Osteogenic Protein-1 on Fibronectin Fragment- and Interleukin-1β-stimulated Matrix Metalloproteinase-13 Expression in Human Chondrocytes* , 2003, Journal of Biological Chemistry.

[33]  J. Pelletier,et al.  The new collagenase, collagenase-3, is expressed and synthesized by human chondrocytes but not by synoviocytes. A role in osteoarthritis. , 1996, The Journal of clinical investigation.

[34]  G. Homandberg,et al.  Cartilage damaging activities of fibronectin fragments derived from cartilage and synovial fluid. , 1998, Osteoarthritis and cartilage.

[35]  G. Homandberg,et al.  Fibronectin fragments cause chondrolysis of bovine articular cartilage slices in culture. , 1992, The Journal of biological chemistry.

[36]  M. Burdick,et al.  Regulation of angiogenesis by the C-X-C chemokines interleukin-8 and epithelial neutrophil activating peptide 78 in the rheumatoid joint. , 2001, Arthritis and rheumatism.

[37]  R. Terkeltaub,et al.  Cartilage and joint inflammation. Regulation of IL-8 expression by human articular chondrocytes. , 1992, Journal of immunology.

[38]  M. Lotz,et al.  Production of the chemokine RANTES by articular chondrocytes and role in cartilage degradation. , 2001, Arthritis and rheumatism.

[39]  R. Loeser,et al.  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 , 2002 .

[40]  K. Nishioka,et al.  Characterization of cells from pannus-like tissue over articular cartilage of advanced osteoarthritis. , 2004, Osteoarthritis and cartilage.