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Regulated expression of the interferon-β gene in mice

Gene Therapy volume 15, pages 1–11 (2008)Cite this article

Abstract

A single plasmid regulated expression vector based upon a mifepristone-inducible two plasmid system, termed pBRES, has been constructed and tested in mice using murine interferon-b (mIFNb) as the transgene. The expression of mIFNb in the circulation was followed by measuring the systemic induction of IP-10, a validated biomarker for mIFNb in mice. Long-term, inducible expression of mIFNb was demonstrated following a single intramuscular (i.m.) injection of the pBRES mIFNb plasmid vector into the hind limb of mice. Induction of mIFNb expression was achieved by administration of the small molecule inducer, mifepristone (MFP). Plasmid DNA and mIFNb mRNA levels in the injected muscles correlated with mIFNb expression as monitored by IP-10 over a 3-month time period. Renewable transgene expression was achieved following repeat administration of the plasmid at 3 months following the first plasmid injection. A dose-dependent increase in expression was demonstrated by varying the amount of injected plasmid or the amount of the inducer administered to the mice. Finally, the pBRES plasmid expressing mIFNb under control of the inducer, MFP, was shown to be efficacious in a murine model of experimental allergic encephalomyelitis, supporting the feasibility of gene-based therapeutic approaches for treating diseases such as multiple sclerosis.

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References

  1. Compston A, oles A . Multiple sclerosis. Lancet 2002; 359: 1221–1231.

    Article  Google Scholar 

  2. Keegan BM, Noseworthy JH . Multiple sclerosis. Annu Rev Med 2002; 53: 258–302.

    Article  Google Scholar 

  3. Steinman L . Multiple sclerosis: a coordinated immunological attack against myelin in the central nervous system. Cell 1996; 85: 299–302.

    Article  CAS  Google Scholar 

  4. Bayas A, Gold R . Lessons from 10 years of interferon beta-1b (Betaferon/Betaseron) treatment. J Neurol 2003; 250 (Suppl 4): IV3–IV8.

    PubMed  Google Scholar 

  5. Malucchi SM, Sala A, Gilli F, Bottero R, Di Sapio A, Capobianco M et al. Neutralizing antibodies reduce the efficacy of IFNb during treatment of multiple sclerosis. Neurology 2004; 62: 2031–2037.

    Article  CAS  Google Scholar 

  6. Triantaphyllopoulos K, Croxford J, Baker D, Chernajovsky Y . Cloning and expression of murine IFNb and a TNF antagonist for gene therapy of experimental allergic encephalomyelitis. Gene Therapy 1998; 5: 253–263.

    Article  CAS  Google Scholar 

  7. Schaefer C, Hidalgo TR, Cashion L, Petry H, Brooks A, Szymanski P et al. Gene-based delivery of interferon-beta is efficacious in a murine model of experimental allergic encephalomyelitis. J Interferon Cytokine Res 2006; 26: 449–454.

    Article  CAS  Google Scholar 

  8. Jaini R, Hannaman D, Johnson JM, Bernard RM, Altuntas CZ, Delasalas MM et al. Gene-based intramuscular interferon-beta therapy for experimental autoimmune encephalomyelitis. Mol Ther 2006; 14: 416–422.

    Article  CAS  Google Scholar 

  9. Baron U, Bujard H . Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. Methods Enzymol 2000; 327: 401–421.

    Article  CAS  Google Scholar 

  10. No D, Yao TP, Evans R . Ecdysone-inducible gene expression in mammalian cells and transgenic mice. Proc Natl Acad Sci USA 1996; 93: 3346–3351.

    Article  CAS  Google Scholar 

  11. Pollock R, Clackson T . Dimerizer-regulated gene expression. Curr Opin Biotechnol 2002; 13: 459–467.

    Article  CAS  Google Scholar 

  12. Burcin MM, Schiedner G, Kochanek S, Tsai SY, O’Malley BW . Adenovirus-mediated regulable target gene expression in vivo. Proc Natl Acad Sci USA 1999; 96: 355–360.

    Article  CAS  Google Scholar 

  13. Nordstrom JL . Antiprogestin-controllable transgene regulation in vivo. Curr Opin Biotech 2002; 13: 453–458.

    Article  CAS  Google Scholar 

  14. Nordstrom JL . The anti-progestin-dependent GeneSwitch® system for regulated gene therapy. Steroids 2003; 68: 1085–1094.

    Article  CAS  Google Scholar 

  15. Szymanski P, Kretschmer PJ, Bauzon M, Jin F, Qian HS, Rubanyi GM et al. Development and validation of a robust and versatile one-plasmid regulated gene expression system. Mol Ther 2007; 15: 1340–1347.

    Article  CAS  Google Scholar 

  16. Petry H, Cashion L, Szymanski P, Ast O, Orme A, Gross C et al. Mx1 and IP-10: biomarkers to measure interferon-beta activity in mice following gene-based delivery. J Interferon Cytokine Res 2006; 26: 699–705.

    Article  CAS  Google Scholar 

  17. Yu M, Nishiyama A, Trapp BD, Tuohy VK . Interferon β inhibits progression of relapsing-remitting experimental autoimmune encephalomyelitis. J Immunol 1996; 64: 91–100.

    CAS  Google Scholar 

  18. Tuohy VK, Yu M, Yin L, Mathisen PM, Johnson JM, Kawczak JA . Modulation of the IL-10/IL-12 cytokine circuit by interferon-beta inhibits the development of epitope spreading and disease progression in murine autoimmune encephalomyelitis. J Neuroimmunol 2000; 111: 55–63.

    Article  CAS  Google Scholar 

  19. Goodin DS . Treatment of multiple sclerosis with human beta interferon. Int MS J 2005; 12: 96–108.

    CAS  PubMed  Google Scholar 

  20. Kappos L, Polman CH, Freedman MS, Edan G, Hartung HP, Miller DH et al. Treatment with interferon beta-1b delays conversion to clinically definite and McDonald MS in patients with clinically isolated syndromes. Neurology 2006; 67: 1242–1249.

    Article  CAS  Google Scholar 

  21. Hyde SC, Southern KW, Gileadi U, Fitzjohn EM, Mofford KA, Waddell BE et al. Repeat administration of DNA/liposomes to the nasal epithelium of patients with cystic fibrosis. Gene Therapy 2000; 7: 1156–1165.

    Article  CAS  Google Scholar 

  22. Prud’homme GJ, Glinka Y, Khan AS, Draghia-Akli R . Electroporation-enhanced nonviral gene transfer for the prevention or treatment of immunological, endocrine and neoplastic diseases. Curr Gene Ther 2006; 6: 243–273.

    Article  Google Scholar 

  23. Wells DJ . Viral and non-viral methods for gene transfer into skeletal muscle. Curr Opin Drug Discov 2006; 9: 163–168.

    CAS  Google Scholar 

  24. Ratanamart J, Shaw JAM . Plasmid-mediated muscle-targeted gene therapy for circulating therapeutic protein replacement: a tale of the tortoise and the hare? Curr Gene Ther 2006; 6: 93–110.

    Article  CAS  Google Scholar 

  25. Sitruk-Ware R, Spitz IM . Pharmacological properties of mifepristone: toxicology and safety in animals and human studies. Contraception 2003; 68: 409–420.

    Article  CAS  Google Scholar 

  26. DeBattista C, Belanoff J . The use of mifepristone in the treatment of neuropsychiatric disorders. Trend Endocrinol Metab 2006; 17: 117–121.

    Article  CAS  Google Scholar 

  27. Majumder S, Zhou LZH, Chaturvedi P, Babcock G, Aras S, Ransohoff RM . Regulation of human IP-10 gene expression in astrocytoma cells by inflammatory cytokines. J Neurosci Res 1998; 54: 169–180.

    Article  CAS  Google Scholar 

  28. Franciotta D, Martino G, Zardini E, Furlan R, Bergamaschi R, Andreoni L et al. Serum and CSF levels of MCP-1 and IP-10 in multiple sclerosis patients with acute and stable disease and undergoing immunomodulatory therapies. J Neuroimmunol 2001; 115: 192–198.

    Article  CAS  Google Scholar 

  29. Buttmann M, Merzyn C, Rieckmann P . Interferon-β induces transient systemic IP-10/CXCL10 chemokine release in patients with multiple sclerosis. J Neuroimmunol 2004; 156: 195–203.

    Article  CAS  Google Scholar 

  30. Hong J, Zang YZQ, Hutton G, Rivera VM, Zhang JZ . Gene expression profiling of relevant biomarkers for treatment evaluation in multiple sclerosis. J Neuroimmunol 2004; 152: 126–139.

    Article  CAS  Google Scholar 

  31. Trollet C, Bloquel C, Scherman D, Bigey P . Electrotransfer into skeletal muscle for protein expression. Curr Gene Ther 2006; 6: 561–578.

    Article  CAS  Google Scholar 

  32. Brooks AR, Wang P, Wu F, Harkins RN . Inflammatory gene expression profile following electroporation enhanced delivery of plasmids to skeletal muscle (submitted for publication).

  33. Yew NS, Zhao H, Prizbylska M, Wu IH, Tousignant JD, Scheule RK et al. CpG-depleted plasmid DNA vectors with enhanced safety and long-term gene expression in vivo. Mol Ther 2002; 5: 731–738.

    Article  CAS  Google Scholar 

  34. Yew NS, Cheng SH . Reducing the immunostimulatory activity of CpG-containing plasmid DNA vectors for non-viral gene therapy. Expert Opin Drug Deliv 2004; 1: 115–125.

    Article  CAS  Google Scholar 

  35. Hodges BL, Taylor KM, Joseph MF, Bourgeoios SA, Scheule RK . Long-term transgene expression from plasmid DNA gene therapy vectors is negatively affected by CpG dinucleotides. Mol Ther 2004; 10: 269–278.

    Article  CAS  Google Scholar 

  36. Gronevik E, von Steyern FV, Kalhovde JM, Tjelle TE, Mathiesen J . Gene expression and immune response kinetics using electroporation-mediated DNA delivery to muscle. J Gene Med 2005; 7: 218–227.

    Article  CAS  Google Scholar 

  37. Abruzzese RV, MacLaughlin FC, Smith LC, Nordstrom JL . Regulated expression of plasmid-based gene therapies. Methods Mol Med 2002; 69: 109–122.

    CAS  PubMed  Google Scholar 

  38. Hardy MP, Owczarek CM, Trajanovska S, Liu X, Kola I, Hertzog PJ . The soluble murine type I interferon receptor Ifnar-2 is present in serum, is independently regulated, and has both agonistic and antagonistic properties. Blood 2001; 97: 473–482.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Perry Liu, Heather Gibson, Kathy White, ToniRose Hidalgo and Vladimir Triantafillidis for animal pharmacology, and other members of the Berlex Gene Technologies, Pharmacology and Systems Biology Departments for their technical assistance and scientific input.

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Author notes

  1. G M Rubanyi

    Present address: 3Current address: Cardium Therapeutics Inc., 3611 Valley Center Drive, San Diego, CA 92130, USA,

Authors and Affiliations

  1. Department of Gene Technologies, Berlex Biosciences, Richmond, CA, USA

    R N Harkins, P Szymanski, H Petry, A Brooks, P J Kretschmer, A Orme, P Wang, G M Rubanyi & T W Hermiston

  2. Department of Pharmacology, Berlex Biosciences, Richmond, CA, USA

    H S Qian & C Schaefer

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  1. R N Harkins
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  2. P Szymanski
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  4. A Brooks
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  5. H S Qian
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  6. C Schaefer
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  7. P J Kretschmer
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  8. A Orme
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  9. P Wang
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Correspondence to R N Harkins.

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Harkins, R., Szymanski, P., Petry, H. et al. Regulated expression of the interferon-β gene in mice. Gene Ther 15, 1–11 (2008). https://doi.org/10.1038/sj.gt.3302998

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  • DOI: https://doi.org/10.1038/sj.gt.3302998

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