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Inhibition of cartilage destruction by double gene transfer of IL-1Ra and IL-10 involves the activin pathway

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Abstract

The objective of the study was to determine the effects and the molecular background of interleukin-1 receptor antagonist (IL-1Ra) and vIL-10 double gene transfer into human synovial fibroblasts from patients with rheumatoid arthritis (RA) using the SCID mouse model for cartilage erosion in RA. RA synovial fibroblasts were transduced with retro- or adenoviruses encoding IL-1Ra and/or viral IL-10 (vIL-10). SCID mice were engrafted subcutaneously with IL-1Ra and vIL-10 transduced human rheumatoid synovial fibroblasts and normal cartilage. In parallel, gene expression analysis before and after gene transfer using RNA arbitrarily primed PCR in combination with cDNA array was performed. vIL-10 and IL-1Ra double gene transfer resulted in inhibition of cartilage invasion and degradation by RA synovial fibroblasts when compared with control transduced and non-transduced implants. Expression of key genes that were altered after double gene transfer were related to the activin pathway. The results demonstrate not only that virus-based gene transfer using a combination of two joint-protective genes is a feasible approach to inhibit cartilage degradation by activated RA synovial fibroblasts, but also that the underlying molecular effects include modulation of the activin pathway.

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References

  1. Gay S, Gay RE, Koopman WJ . Molecular and cellular mechanisms of joint destruction in rheumatoid arthritis: two cellular mechanisms explain joint destruction? Ann Rheum Dis 1993 52: S39–S47

    Article  PubMed  PubMed Central  Google Scholar 

  2. Firestein GS . Invasive fibroblast-like synoviocytes in rheumatoid arthritis. Passive responders or transformed aggressors? Arthritis Rheum 1996 39: 1781–1790

    Article  CAS  PubMed  Google Scholar 

  3. Pap T, Müller-Ladner U, Gay RE, Gay S . Fibroblast biology. Role of synovial fibroblasts in the pathogenesis of rheumatoid arthritis Arthritis Res 2000 2: 361–367

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Müller-Ladner U, Gay RE, Gay S . Molecular biology of cartilage and bone destruction Curr Opin Rheumatol 1998 10: 212–219

    Article  PubMed  Google Scholar 

  5. Müller-Ladner U et al. Synovial fibroblasts of patients with rheumatoid arthritis attach to and invade normal human cartilage when engrafted into SCID mice Am J Pathol 1996 149: 1607–1615

    PubMed  PubMed Central  Google Scholar 

  6. Müller-Ladner U, Kriegsmann J, Gay RE, Gay S . Oncogenes in rheumatoid arthritis Rheum Dis Clin North Am 1995 21: 675–690

    Article  PubMed  Google Scholar 

  7. Keyszer GM et al. Comparative analysis of cathepsin L, cathepsin D, and collagenase messenger RNA expression in synovial tissues of patients with rheumatoid arthritis and osteoarthritis, by in situ hybridization Arthritis Rheum 1995 38: 976–984

    Article  CAS  PubMed  Google Scholar 

  8. Pap T et al. Activation of synovial fibroblasts in rheumatoid arthritis: lack of expression of the tumour suppressor PTEN at sites of invasive growth and destruction Arthritis Res 2000 2: 59–64

    Article  CAS  PubMed  Google Scholar 

  9. Grimbacher B, Aicher WK, Peter HH, Eibel H . Measurement of transcription factor c-fos and EGR-1 mRNA transcription levels in synovial tissue by quantitative RT-PCR Rheumatol Int 1997 17: 109–112

    Article  CAS  PubMed  Google Scholar 

  10. Arend WP, Dayer JM . Inhibition of the production and effects of interleukin-1 and tumor necrosis factor alpha in rheumatoid arthritis Arthritis Rheum 1995 38: 151–160

    Article  CAS  PubMed  Google Scholar 

  11. Probert L, Plows D, Kontogeorgos G, Kollias G . The type I interleukin-1 receptor acts in series with tumor necrosis factor (TNF) to induce arthritis in TNF-transgenic mice Eur J Immunol 1995 25: 1794–1797

    Article  CAS  PubMed  Google Scholar 

  12. van de Loo FA et al. Role of interleukin-1, tumor necrosis factor alpha, and interleukin-6 in cartilage proteoglycan metabolism and destruction. Effect of in situ blocking in murine ant Arthritis Rheum 1995 38: 164–172

    Article  CAS  PubMed  Google Scholar 

  13. Kalden JR . How do the biologics fit into the current DMARD armamentarium? J Rheumatol 2001 62: 27–35

    CAS  Google Scholar 

  14. Elliott MJ et al. Randomised double-blind comparison of chimeric monoclonal antibody to tumour necrosis factor alpha (cA2) versus placebo in rheumatoid arthritis Lancet 1994 344: 1105–1110

    Article  CAS  PubMed  Google Scholar 

  15. Lipsky PE et al. Infliximab and methotrexate in the treatment of rheumatoid arthritis. Anti-tumor necrosis factor trial in rheumatoid arthritis with concomitant therapy study group N Engl J Med 2000 343: 1594–1602

    Article  CAS  PubMed  Google Scholar 

  16. Arend WP et al. Binding of IL-1 alpha, IL-1 beta and IL-1 receptor antagonist by soluble IL-1 receptors and levels of soluble IL-1 receptors in synovial fluids J Immunol 1994 153: 4766–4774

    CAS  PubMed  Google Scholar 

  17. Moreland LW et al. Recombinant soluble tumor necrosis factor receptor (p80) fusion protein: toxicity and dose finding trial in refractory rheumatoid arthritis J Rheumatol 1996 23: 1849–1855

    CAS  PubMed  Google Scholar 

  18. Arend WP . Interleukin-1 receptor antagonist Adv Immunol 1993 54: 167–227

    Article  CAS  PubMed  Google Scholar 

  19. Campion GV et al. Dose-range and dose-frequency study of recombinant human interleukin-1 receptor antagonist in patients with rheumatoid arthritis. The IL-1Ra Arthritis Study Group Arthritis Rheum 1996 39: 1092–1101

    Article  CAS  PubMed  Google Scholar 

  20. Bresnihan B et al. Treatment of rheumatoid arthritis with recombinant human interleukin-1 receptor antagonist Arthritis Rheum 1998 41: 2196–2204

    Article  CAS  PubMed  Google Scholar 

  21. Eisenberg SP et al. Primary structure and functional expression from complementary DNA of a human interleukin-1 receptor antagonist Nature 1990 343: 341–346

    Article  CAS  PubMed  Google Scholar 

  22. Bresnihan B . The safety and efficacy of interleukin-1 receptor antagonist in the treatment of rheumatoid arthritis Semin Arthritis Rheum 2001 30: 17–20

    Article  CAS  PubMed  Google Scholar 

  23. Roux-Lombard P et al. Cytokines, metalloproteinases, their inhibitors and cartilage oligomeric matrix protein: relationship to radiological progression and inflammation in early rheumatoid arthritis. A prospective 5-year study Rheumatology 2001 40: 544–551

    Article  CAS  PubMed  Google Scholar 

  24. Watt I, Cobby M . Treatment of rheumatoid arthritis patients with interleukin-1 receptor antagonist: radiologic assessment Semin Arthritis Rheum 2001 30: 21–25

    Article  CAS  PubMed  Google Scholar 

  25. Isomaki P, Luukkainen R, Toivanen P, Punnonen J . The presence of interleukin-13 in rheumatoid synovium and its antiinflammatory effects on synovial fluid macrophages from patients with rheumatoid arthritis Arthritis Rheum 1996 39: 1693–1702

    Article  CAS  PubMed  Google Scholar 

  26. Katsikis PD et al. Immunoregulatory role of interleukin 10 in rheumatoid arthritis J Exp Med 1994 179: 1517–1527

    Article  CAS  PubMed  Google Scholar 

  27. Miossec P et al. Low levels of interleukin-4 and high levels of transforming growth factor beta in rheumatoid synovitis Arthritis Rheum 1990 33: 1180–1187

    Article  CAS  PubMed  Google Scholar 

  28. Moore KW et al. Homology of cytokine synthesis inhibitory factor (IL-10) to the Epstein-Barr virus gene BCRFI Science 1990 248: 1230–1234

    Article  CAS  PubMed  Google Scholar 

  29. Suzuki T et al. Viral interleukin 10 (IL-10), the human herpes virus 4 cellular IL-10 homologue, induces local anergy to allogeneic and syngeneic tumors J Exp Med 1995 182: 477–486

    Article  CAS  PubMed  Google Scholar 

  30. MacNeil IA et al. IL-10, a novel growth cofactor for mature and immature T cells J Immunol 1990 145: 4167–4173

    CAS  PubMed  Google Scholar 

  31. Go NF et al. Interleukin 10, a novel B cell stimulatory factor: unresponsiveness of X chromosome-linked immunodeficiency B cells J Exp Med 1990 172: 1625–1631

    Article  CAS  PubMed  Google Scholar 

  32. Lacraz S et al. IL-10 inhibits metalloproteinase and stimulates TIMP-1 production in human mononuclear phagocytes J Clin Invest 1995 96: 2304–2310

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. de Waal MR et al. Interleukin 10 (IL-10) and viral IL-10 strongly reduce antigen-specific human T cell proliferation by diminishing the antigen-presenting capacity of monocytes via downregulation of class II major histocompatibility complex expression J Exp Med 1991 174: 915–924

    Article  Google Scholar 

  34. de Waal MR et al. Interleukin 10(IL-10) inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes J Exp Med 1991 174: 1209–1220

    Article  Google Scholar 

  35. Evans CH et al. Clinical trial to assess the safety, feasibility, and efficacy of transferring a potentially anti-arthritic cytokine gene to human joints with rheumatoid arthritis Hum Gene Ther 1996 7: 1261–1280

    Article  CAS  PubMed  Google Scholar 

  36. Müller-Ladner U et al. Human IL-1Ra gene transfer into human synovial fibroblasts is chondroprotective J Immunol 1997 158: 3492–3498

    PubMed  Google Scholar 

  37. Müller-Ladner U et al. Gene transfer of cytokine inhibitors into human synovial fibroblasts in the SCID mouse model Arthritis Rheum 1999 42: 490–497

    Article  PubMed  Google Scholar 

  38. Van Den Berg WB, Joosten LA, Helsen M, Van De Loo FA . Amelioration of established murine collagen-induced arthritis with anti- IL-1 treatment Clin Exp Immunol 1994 95: 237–243

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Williams RO, Feldmann M, Maini RN . Anti-tumor necrosis factor ameliorates joint disease in murine collagen-induced arthritis Proc Natl Acad Sci USA 1992 89: 9784–9788

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Joosten LA et al. Role of interleukin-4 and interleukin-10 in murine collagen-induced arthritis. Protective effect of interleukin-4 and interleukin-10 treatment on cartilage destruction Arthritis Rheum 1997 40: 249–260

    Article  CAS  PubMed  Google Scholar 

  41. Evans CH et al. Transfer and intra-articular expression of the IL-1Ra cDNA in human rheumatoid joints Arthritis Res 2001 3: P33

    Article  PubMed Central  Google Scholar 

  42. Neumann E et al. Additive cartilage protection by retrovirus-based double-gene transfer with IL-1ra and IL-10 in the SCID mouse model for RA Arthritis Rheum 2001 43: S170

    Google Scholar 

  43. Gabay C et al. Increased production of intracellular interleukin-1 receptor antagonist type I in the synovium of mice with collagen-induced arthritis: a possible role in the resolution of arthritis Arthritis Rheum 2001 44: 451–462

    Article  CAS  PubMed  Google Scholar 

  44. Seitz M et al. Interleukin-10 differentially regulates cytokine inhibitor and chemokine release from blood mononuclear cells and fibroblasts Eur J Immunol 1995 25: 1129–1132

    Article  CAS  PubMed  Google Scholar 

  45. Yu J, Dolter KE . Production of activin A and its roles in inflammation and hematopoiesis Cytokines Cell Mol Ther 1997 3: 169–177

    CAS  PubMed  Google Scholar 

  46. Mathews LS, Vale WW . Molecular and functional characterization of activin receptors Receptor 1993 3: 173–181

    CAS  PubMed  Google Scholar 

  47. Mather JP . Follistatins and alpha 2-macroglobulin are soluble binding proteins for inhibin and activin Horm Res 1996 45: 207–210

    Article  CAS  PubMed  Google Scholar 

  48. Ying SY . Inhibins, activins, and follistatins: gonadal proteins modulating the secretion of follicle-stimulating hormone Endocr Rev 1988 9: 267–293

    Article  CAS  PubMed  Google Scholar 

  49. de Kretser DM, Hedger MP, Phillips DJ . Activin A and follistatin: their role in the acute phase reaction and inflammation J Endocrinol 1999 161: 195–198

    Article  CAS  PubMed  Google Scholar 

  50. Danila DC et al. Activin effects on neoplastic proliferation of human pituitary tumors J Clin Endocrinol Metab 2000 85: 1009–1015

    CAS  PubMed  Google Scholar 

  51. Schubert D et al. Activin is a nerve cell survival molecule Nature 1990 344: 868–870

    Article  CAS  PubMed  Google Scholar 

  52. Hashimoto M et al. Activin/EDF as an inhibitor of neural differentiation Biochem Biophys Res Commun 1990 173: 193–200

    Article  CAS  PubMed  Google Scholar 

  53. Hübner G, Alzheimer C, Werner S . Activin: a novel player in tissue repair processes Histol Histopathol 1999 14: 295–304

    PubMed  Google Scholar 

  54. Ohga E, Matsuse T, Teramoto S, Ouchi Y . Activin receptors are expressed on human lung fibroblast and activin A facilitates fibroblast-mediated collagen gel contraction Life Sci 2000 66: 1603–1613

    Article  CAS  PubMed  Google Scholar 

  55. Kozian DH, Ziche M, Augustin HG . The activin-binding protein follistatin regulates autocrine endothelial cell activity and induces angiogenesis Lab Invest 1997 76: 267–276

    CAS  PubMed  Google Scholar 

  56. Kaneko H et al. Direct stimulation of osteoclastic bone resorption by bone morphogenetic protein (BMP)-2 and expression of BMP receptors in mature osteoclasts Bone 2000 27: 479–486

    Article  CAS  PubMed  Google Scholar 

  57. Hummel KM et al. Cysteine proteinase cathepsin K mRNA is expressed in synovium of patients with rheumatoid arthritis and is detected at sites of synovial bone destruction J Rheumatol 1998 25: 1887–1894

    CAS  PubMed  Google Scholar 

  58. Funaba M et al. Follistatin and activin in bone: expression and localization during endochondral bone development Endocrinology 1996 137: 4250–4259

    Article  CAS  PubMed  Google Scholar 

  59. Yonemori K et al. Bone morphogenetic protein receptors and activin receptors are highly expressed in ossified ligament tissues of patients with ossification of the posterior longitudinal ligament Am J Pathol 1997 150: 1335–1347

    CAS  PubMed  PubMed Central  Google Scholar 

  60. Miyazono K, Kusanagi K, Inoue H . Divergence and convergence of TGF-beta/BMP signaling J Cell Physiol 2001 187: 265–276

    Article  CAS  PubMed  Google Scholar 

  61. Gribi R, Tanaka T, Harper-Summers R, Yu J . Expression of activin A in inflammatory arthropathies Mol Cell Endocrinol 2001 180: 163–167

    Article  CAS  PubMed  Google Scholar 

  62. Yu EW, Dolter KE, Shao LE, Yu J . Suppression of IL-6 biological activities by activin A and implications for inflammatory arthropathies Clin Exp Immunol 1998 112: 126–132

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Sugiyama M et al. Expression of activin A is increased in cirrhotic and fibrotic rat livers Gastroenterology 1998 114: 550–558

    Article  CAS  PubMed  Google Scholar 

  64. Nonomura Y et al. Suppression of arthritis by forced expression of cyclin-dependent kinase inhibitor p21(Cip1) gene into the joints Int Immunol 2001 13: 723–731

    Article  CAS  PubMed  Google Scholar 

  65. Taniguchi K et al. Induction of the p16INK4a senescence gene as a new therapeutic strategy for the treatment of rheumatoid arthritis Nat Med 1999 5: 760–767

    Article  CAS  PubMed  Google Scholar 

  66. Nasu K et al. Adenoviral transfer of cyclin-dependent kinase inhibitor genes suppresses collagen-induced arthritis in mice J Immunol 2000 165: 7246–7252

    Article  CAS  PubMed  Google Scholar 

  67. Müller-Ladner U et al. Activation of the IL-4 STAT pathway in rheumatoid synovium J Immunol 2000 164: 3894–3901

    Article  PubMed  Google Scholar 

  68. Li WQ, Dehnade F, Zafarullah M . Oncostatin M-induced matrix metalloproteinase and tissue inhibitor of metalloproteinase-3 genes expression in chondrocytes requires Janus kinase/STAT signaling pathway J Immunol 2001 166: 3491–3498

    Article  CAS  PubMed  Google Scholar 

  69. Zauberman A, Oren M, Zipori D . Involvement of p21(WAF1/Cip1), CDK4 and Rb in activin A mediated signaling leading to hepatoma cell growth inhibition Oncogene 1997 15: 1705–1711

    Article  CAS  PubMed  Google Scholar 

  70. Wang M et al. Growth of HPV-18 immortalized human prostatic intraepithelial neoplasia cell lines. Influence of IL-10, follistatin, activin-A and DHT Int J Oncol 1999 14: 1185–1195

    CAS  PubMed  Google Scholar 

  71. Garkavtsev I et al. The candidate tumour suppressor p33ING1 cooperates with p53 in cell growth control Nature 1998 391: 295–298

    Article  CAS  PubMed  Google Scholar 

  72. Van Roon JA, Lafeber FP, Bijlsma JW . Synergistic activity of interleukin-4 and interleukin-10 in suppression of inflammation and joint destruction in rheumatoid arthritis Arthritis Rheum 2001 44: 3–12

    Article  CAS  PubMed  Google Scholar 

  73. Liu J et al. Anti-sense oligonucleotide of p21(waf1/cip1) prevents interleukin 4-mediated elevation of p27(kip1) in low grade astrocytoma cells Oncogene 2000 19: 661–669

    Article  CAS  PubMed  Google Scholar 

  74. O'Farrell AM et al. Stat3-dependent induction of p19INK4D by IL-10 contributes to inhibition of macrophage proliferation J Immunol 2000 164: 4607–4615

    Article  CAS  PubMed  Google Scholar 

  75. Tanaka M et al. Cloning of follistatin-related protein as a novel autoantigen in systemic rheumatic diseases Int Immunol 1998 10: 1305–1314

    Article  CAS  PubMed  Google Scholar 

  76. Zucker S et al. Tumorigenic potential of extracellular matrix metalloproteinase inducer Am J Pathol 2001 158: 1921–1928

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Pap T, Müller-Ladner U, Gay R, Gay S . Gene therapy in rheumatoid arthritis: how to target joint destruction? Arthritis Res 1999 1: 5–9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. Arnett FC et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis Arthritis Rheum 1988 31: 315–324

    Article  CAS  PubMed  Google Scholar 

  79. Lechman ER et al. Direct adenoviral gene transfer of viral IL-10 to rabbit knees with experimental arthritis ameliorates disease in both injected and contralateral control knees J Immunol 1999 163: 2202–2208

    CAS  PubMed  Google Scholar 

  80. Danos O, Mulligan RC . Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges Proc Natl Acad Sci USA 1988 85: 6460–6464

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Baragi VM et al. Technology evaluation: MFG-IRAP, University of Pittsburgh Curr Opin Mol Ther 2000 2: 216–220

    CAS  PubMed  Google Scholar 

  82. Makarov SS et al. Suppression of experimental arthritis by gene transfer of interleukin 1 receptor antagonist cDNA Proc Natl Acad Sci USA 1996 93: 402–406

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Kullmann F et al. Kinesin-like protein CENP-E is upregulated in rheumatoid synovial fibroblasts Arthritis Res 1999 1: 71–80

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. McClelland M, Welsh J . RNA fingerprinting by arbitrarily primed PCR PCR Meth Appl 1994 4: S66–S81

    Article  CAS  Google Scholar 

  85. Judex M et al. ‘Inverse wrap’ - an improved implantation technique for virus-transduced synovial fibroblasts in the SCID mouse model for rheumatoid arthritis (RA) Mod Rheum 2001 11: 145–150

    Article  CAS  Google Scholar 

  86. Morrison TB, Weis JJ, Wittwer CT . Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification Biotechniques 1998 24: 954–958

    CAS  PubMed  Google Scholar 

  87. Higuchi R, Fockler C, Dollinger G, Watson R . Kinetic PCR analysis: real-time monitoring of DNA amplification reactions Biotechnology (NY) 1993 11: 1026–1030

    CAS  Google Scholar 

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Acknowledgements

This study was supported by grants of the German Academic Research Society (DFG Mu 1383/3-1, 1383/3-3, and Ku 1024/6-1), as well as by the Swiss National Foundation (32-64142.00). The authors wish to thank Birgit Riepl and Wibke Ballhorn for excellent technical assistance.

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Authors and Affiliations

  1. Department of Internal Medicine I, University of Regensburg, Germany

    E Neumann, M Judex, F Kullmann, J Schölmerich & U Müller-Ladner

  2. Department of Orthopedics, University of Regensburg, Germany

    J Grifka

  3. Center for Molecular Orthopedics, Harvard Medical School, Boston, MA, USA

    P D Robbins

  4. Department of Rheumatology, Center for Experimental Rheumatology, University Hospital Zürich, Switzerland

    T Pap, R E Gay & S Gay

  5. University of Pittsburgh, School of Medicine, Pittsburgh, PA, USA

    C H Evans

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  1. E Neumann
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Neumann, E., Judex, M., Kullmann, F. et al. Inhibition of cartilage destruction by double gene transfer of IL-1Ra and IL-10 involves the activin pathway. Gene Ther 9, 1508–1519 (2002). https://doi.org/10.1038/sj.gt.3301811

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