Bone morphogenetic proteins 2 and 6, expressed in arthritic synovium, are regulated by proinflammatory cytokines and differentially modulate fibroblast-like synoviocyte apoptosis.

OBJECTIVE To examine the expression, regulation, and potential roles of bone morphogenetic proteins (BMPs) in arthritic synovium. METHODS Expression of BMPs in arthritic synovium from patients with rheumatoid arthritis (RA) or spondylarthropathy (SpA) and in noninflamed synovium from patients undergoing diagnostic or therapeutic arthroscopies was studied by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, immunohistochemistry, and 2-color immunofluorescence. In vitro regulation of gene expression in fibroblast-like synoviocytes (FLS) was determined by real-time quantitative RT-PCR and immunohistochemistry. We used (3)H-thymidine incorporation after serum deprivation-induced growth arrest to examine effects on FLS proliferation. FLS apoptosis was evaluated by flow cytometry cell cycle analysis. Apoptotic cells in synovial tissue were detected by TUNEL staining. RESULTS Transcripts of different BMPs, most strikingly BMP-2 and BMP-6, were detected in synovial tissues. By Western blot, BMP-2 and BMP-6 precursor protein was found in RA and SpA synovial tissue extracts, but not in extracts of noninflamed synovial tissue. By immunohistochemistry, BMP-2 and BMP-6 were detected in the hyperplastic lining and the sublining layer of synovium from RA and SpA patients, both in CD90+ cells (FLS) and in some CD68+ cells (macrophages). Proinflammatory cytokines, such as interleukin-1beta and tumor necrosis factor alpha, but not interferon-gamma, enhanced the expression of BMP-2 and BMP-6 transcripts in FLS in vitro. Neither BMP-2 nor BMP-6 affected FLS proliferation. BMP-2 promoted FLS apoptosis, whereas BMP-6 protected against nitric oxide-induced FLS apoptosis. BMP-2-positive apoptotic cells were found in arthritic synovium. CONCLUSION BMP-2 and BMP-6 are expressed in arthritic synovium and are strongly up-regulated by proinflammatory cytokines. Although BMP signaling has been proposed to be involved in cartilage and bone repair in arthritis, this pathway may be equally important in modulating FLS cell populations in inflamed synovium.

[1]  W. B. van den Berg,et al.  Inhibition of Endogenous TGF-β During Experimental Osteoarthritis Prevents Osteophyte Formation and Impairs Cartilage Repair , 2002, The Journal of Immunology.

[2]  H. Zeidler,et al.  Treatment of active ankylosing spondylitis with infliximab: a randomised controlled multicentre trial , 2002, The Lancet.

[3]  G. Firestein,et al.  How important are T cells in chronic rheumatoid synovitis?: II. T cell-independent mechanisms from beginning to end. , 2002, Arthritis and rheumatism.

[4]  F. Luyten,et al.  Skeletal tissue engineering: opportunities and challenges. , 2001, Best practice & research. Clinical rheumatology.

[5]  C. Y. Wang,et al.  Suppression of Tumor Necrosis Factor-mediated Apoptosis by Nuclear Factor κB-independent Bone Morphogenetic Protein/Smad Signaling* , 2001, The Journal of Biological Chemistry.

[6]  P. Klimiuk,et al.  T Cell Activation in Rheumatoid Synovium Is B Cell Dependent1 , 2001, The Journal of Immunology.

[7]  J. Cadet,et al.  Bone Morphogenetic Protein-6 Reduces Ischemia-Induced Brain Damage in Rats , 2001, Stroke.

[8]  Y. Fujio,et al.  Bone Morphogenetic Protein-2 Inhibits Serum Deprivation-induced Apoptosis of Neonatal Cardiac Myocytes through Activation of the Smad1 Pathway* , 2001, The Journal of Biological Chemistry.

[9]  F. Emmrich,et al.  Mosaic chromosomal aberrations in synovial fibroblasts of patients with rheumatoid arthritis, osteoarthritis, and other inflammatory joint diseases , 2001, Arthritis research.

[10]  J. Lemonnier,et al.  Bone Morphogenetic Protein-2 Promotes Osteoblast Apoptosis through a Smad-independent, Protein Kinase C-dependent Signaling Pathway* , 2001, The Journal of Biological Chemistry.

[11]  F. Luyten,et al.  Multipotent mesenchymal stem cells from adult human synovial membrane. , 2001, Arthritis and rheumatism.

[12]  G. Panayi,et al.  Pathogenesis of rheumatoid arthritis. The role of T cells and other beasts. , 2001, Rheumatic diseases clinics of North America.

[13]  C. Deng,et al.  TGF-β/Smad3 Signals Repress Chondrocyte Hypertrophic Differentiation and Are Required for Maintaining Articular Cartilage , 2001, The Journal of cell biology.

[14]  Xu Cao,et al.  Transcriptional Mechanisms of Bone Morphogenetic Protein-induced Osteoprotegrin Gene Expression* , 2001, The Journal of Biological Chemistry.

[15]  Frank Emmrich,et al.  Isolation and characterization of rheumatoid arthritis synovial fibroblasts from primary culture — primary culture cells markedly differ from fourth-passage cells , 2000, Arthritis research.

[16]  M. Kizaki,et al.  Bone morphogenetic protein-2 induces apoptosis in human myeloma cells with modulation of STAT3. , 2000, Blood.

[17]  N. Zvaifler,et al.  Mesenchymal cells expressing bone morphogenetic protein receptors are present in the rheumatoid arthritis joint. , 2000, Arthritis and rheumatism.

[18]  Hiroshi Shibuya,et al.  BMP2-induced Apoptosis Is Mediated by Activation of the TAK1-p38 Kinase Pathway That Is Negatively Regulated by Smad6* , 2000, The Journal of Biological Chemistry.

[19]  K. Miyazono,et al.  Characterization of bone morphogenetic protein-6 signaling pathways in osteoblast differentiation. , 1999, Journal of cell science.

[20]  K. Yamato,et al.  Bone morphogenetic protein-2 enhances osteoclast formation mediated by interleukin-1alpha through upregulation of osteoclast differentiation factor and cyclooxygenase-2. , 1999, Biochemical and biophysical research communications.

[21]  J. Dick,et al.  Bone Morphogenetic Proteins Regulate the Developmental Program of Human Hematopoietic Stem Cells , 1999, The Journal of experimental medicine.

[22]  D. Hu,et al.  Common Molecular Pathways in Skeletal Morphogenesis and Repair , 1998, Annals of the New York Academy of Sciences.

[23]  B. Riggs,et al.  Osteoprotegerin production by human osteoblast lineage cells is stimulated by vitamin D, bone morphogenetic protein-2, and cytokines. , 1998, Biochemical and biophysical research communications.

[24]  F. Breedveld,et al.  Therapeutic efficacy of multiple intravenous infusions of anti-tumor necrosis factor alpha monoclonal antibody combined with low-dose weekly methotrexate in rheumatoid arthritis. , 1998, Arthritis and rheumatism.

[25]  W. B. van den Berg,et al.  Differential effects of local application of BMP-2 or TGF-beta 1 on both articular cartilage composition and osteophyte formation. , 1998, Osteoarthritis and cartilage.

[26]  E. Pease,et al.  Induction of bone morphogenetic protein-2 by interleukin-1 in human fibroblasts. , 1998, Biochemical and biophysical research communications.

[27]  F. Luyten,et al.  Presence of cartilage-derived morphogenetic proteins in articular cartilage and enhancement of matrix replacement in vitro. , 1998, Arthritis and rheumatism.

[28]  Y. Gañán,et al.  Role of BMP-2 and OP-1 (BMP-7) in programmed cell death and skeletogenesis during chick limb development. , 1997, Development.

[29]  G. Firestein.  Invasive fibroblast-like synoviocytes in rheumatoid arthritis. Passive responders or transformed aggressors? , 1996, Arthritis and rheumatism.

[30]  B. Hogan.  Bone morphogenetic proteins in development. , 1996, Current opinion in genetics & development.

[31]  中瀬 尚長.  Transient and localized expression of bone morphogenetic protein-4 messenger RNA during fracture healing , 1995 .

[32]  A. Graham,et al.  The signalling molecule BMP4 mediates apoptosis in the rhombencephalic neural crest , 1994, Nature.

[33]  V. Paralkar,et al.  Osteogenin and recombinant bone morphogenetic protein 2B are chemotactic for human monocytes and stimulate transforming growth factor beta 1 mRNA expression. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[34]  M. Dougados,et al.  The European Spondylarthropathy Study Group preliminary criteria for the classification of spondylarthropathy. , 1991, Arthritis and rheumatism.

[35]  F. Arnett.  Revised criteria for the classification of rheumatoid arthritis. , 1990, Orthopedic nursing.

[36]  V. Rosen,et al.  Novel regulators of bone formation: molecular clones and activities. , 1988, Science.

[37]  M. Liang,et al.  The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. , 1988, Arthritis and rheumatism.

[38]  R. Moskowitz,et al.  Development of criteria for the classification and reporting of osteoarthritis. Classification of osteoarthritis of the knee. Diagnostic and Therapeutic Criteria Committee of the American Rheumatism Association. , 1986, Arthritis and rheumatism.

[39]  M. Urist,et al.  Bone: Formation by Autoinduction , 1965, Science.

[40]  F. Luyten,et al.  Bone morphogenetic proteins and the synovial joints , 2002 .

[41]  J. Massagué.  How cells read TGF-beta signals. , 2000, Nature reviews. Molecular cell biology.

[42]  D. Elewaut,et al.  Needle Arthroscopy of the Knee with Synovial Biopsy Sampling: Technical Experience in 150 Patients , 1999, Clinical Rheumatology.

[43]  J. Massagué.  TGF-beta signal transduction. , 1998, Annual review of biochemistry.

[44]  V. Rosen,et al.  Bone morphogenetic protein and bone morphogenetic protein gene family in bone formation and repair. , 1998, Clinical orthopaedics and related research.