Abstract
The use of gene transfer techniques has become of utmost importance both for the analysis of molecular pathways of rheumatic joint destruction and for the evaluation of novel therapeutic concepts to treat rheumatic diseases. However, gene transfer into synovial fibroblasts faces several challenges, which result mainly from the lack of specific surface markers and the low-proliferation rate of these cells. This chapter describes both nonviral and viral strategies of transferring gene constructs into synovial fibroblasts. It focuses on the use of lipofection for the gene transfer of siRNA to synovial fibroblasts and the use of AMAXA-nucleofection for the nonviral transfer of gene expression constructs. In addition, retro- and lentiviral strategies of gene transfer are introduced. Finally, the SCID mouse in vivo model of rheumatoid joint destruction is described as a means of evaluating the effects of gene transfer on the invasiveness of synovial fibroblasts.
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Kinne, R. W., Emmrich, F., Bail, H., et al. (1995) Expression of activation markers of the rheumatoid arthritis synovial membrane: comment on the article by Qu et al. Arthritis Rheum. 38, 1346–1348.
Mohr, W., Hummler, N., Pelster, B., and Wessinghage, D. (1986) Proliferation of pannus tissue cells in rheumatoid arthritis. Rheumatol. Int. 6, 127–132.
Nykanen, P., Bergroth, V., Raunio, P., Nordstrom, D., and Konttinen, Y. T. (1986) Phenotypic characterization of 3H-thymidine incorporating cells in rheumatoid arthritis synovial membrane. Rheumatol. Int. 6, 269–271.
Baier, A., Meineckel, I., Gay, S., and Pap, T. (2003) Apoptosis in rheumatoid arthritis. Curr. Opin. Rheumatol. 15, 274–279.
Franz, J. K., Pap, T., Hummel, K. M., et al. (2000) Expression of sentrin, a novel antiapoptotic molecule, at sites of synovial invasion in rheumatoid arthritis. Arthritis Rheum. 43, 599–607.
Geiler, T., Kriegsmann, J., Keyszer, G. M., Gay, R. E., and Gay, S. (1994) A new model for rheumatoid arthritis generated by engraftment of rheumatoid synovial tissue and normal human cartilage into SCID mice. Arthritis Rheum. 37, 1664–1671.
Muller-Ladner, U., Kriegsmann, J., Franklin, B. N., et al. (1996) Synovial fibroblasts of patients with rheumatoid arthritis attach to and invade normal human cartilage when engrafted into SCID mice. Am. J. Pathol. 149, 1607–1615.
Pap, T., Muller-Ladner, U., Gay, R. E., and Gay, S. (2000) Fibroblast biology. Role of synovial fibroblasts in the pathogenesis of rheumatoid arthritis. Arthritis Res. 2, 361–367.
Pap, T., Aupperle, K. R., Gay, S., Firestein, G. S., and Gay, R. E. (2001) Invasiveness of synovial fibroblasts is regulated by p53 in the SCID mouse in vivo model of cartilage invasion. Arthritis Rheum. 44, 676–681.
Lenz, P., Bacot, S. M., Frazier-Jessen, M. R., and Feldman, G. M. (2003) Nucleoporation of dendritic cells: efficient gene transfer by electroporation into human monocyte-derived dendritic cells. FEBS Lett. 538, 149–154.
Hamm, A., Krott, N., Breibach, I., Blindt, R., and Bosserhoff, A. K. (2002) Efficient transfection method for primary cells. Tissue Eng 8, 235–245.
Maasho, K., Marusina, A., Reynolds, N. M., Coligan, J. E., and Borrego, F. (2004) Efficient gene transfer into the human natural killer cell line, NKL, using the Amaxa nucleofection system. J. Immunol. Methods 284, 133–140.
Gouze, J. N., Stoddart, M. J., Gouze, E., Palmer, G. D., Ghivizzani, S. C., Grodzinsky, A. J., and Evans, C. H. (2004) In vitro gene transfer to chondrocytes and synovial fibroblasts by adenoviral vectors. Methods Mol. Med. 100, 147–164.
Tonini, T., Claudio, P. P., Giordano, A., and Romano, G. (2004) Transient production of retroviral-and lentiviral-based vectors for the transduction of Mammalian cells. Methods Mol. Biol. 285, 141–148.
Aicher, W. K., Heer, A. H., Trabandt, A., et al. (1994) Over-expression of zinc-finger transcription factor Z-225/Egr-1 in synoviocytes from rheumatoid arthritis patients. J. Immunol. 152, 5940–5948.
Naldini, L. and Verma, I. M. (2000) Lentiviral vectors. Adv. Virus Res. 55, 599–609.
Naldini, L. (1998) Lentiviruses as gene transfer agents for delivery to non-dividing cells. Curr. Opin. Biotechnol. 9, 457–463.
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© 2007 Humana Press Inc., Totowa, NJ
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Meinecke, I., Rutkauskaite, E., Cinski, A., Müller-Ladner, U., Gay, S., Pap, T. (2007). Gene Transfer to Synovial Fibroblast. In: Cope, A.P. (eds) Arthritis Research. Methods in Molecular Medicine, vol 135. Humana Press. https://doi.org/10.1007/978-1-59745-401-8_26
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DOI: https://doi.org/10.1007/978-1-59745-401-8_26
Publisher Name: Humana Press
Print ISBN: 978-1-58829-344-2
Online ISBN: 978-1-59745-401-8
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