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Efficacy of adenoviral TNFα antisense is enhanced by a macrophage specific promoter

Gene Therapy volume 8, pages 223–231 (2001)Cite this article

Abstract

Macrophage-derived TNFα is a critical mediator of inflammation and destruction in diseases such as rheumatoid arthritis and Crohn's disease. These studies were undertaken to develop an effective adenovirus-based strategy to specifically suppress TNFα in primary human macrophages. A variety of promoters and LTRs were evaluated for effective expression in the macrophage cell line RAW 264.7. The CMV promoter and the Visna LTR were the most strongly expressed and were therefore used to drive the expression of TNFα antisense fragments. In transient transfection assays, the antisense fragment terminating at the 3′ end of the first exon (216 bp) was superior to the others (70 and 750 bp), when expressed under the control of either the CMV promoter or the Visna LTR. Adenoviral vectors expressing the 216 bp TNFα antisense fragment, controlled by the CMV promoter or the Visna LTR, were both effective at suppressing LPS-induced TNFα secretion by primary human macrophages. However, the Visna LTR was more effective not only at suppressing LPS-induced TNFα secretion, but also IL-6, which is highly sensitive to TNFα secretion. These results demonstrate that effective, specific, suppression of TNFα in macrophages is possible, employing a directed antisense approach and a promoter system that is highly efficient in human macrophages.

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References

  1. Feldmann M, Brennan FM, Maini RN . Role of cytokines in rheumatoid arthritis Annu Rev Immunol 1996 14: 397–440

    Article  CAS  Google Scholar 

  2. Butler DM, Maini RN, Feldmann M, Brennan FM . Modulation of proinflammatory cytokine release in rheumatoid synovial membrane cell cultures. Comparison of monoclonal anti-TNF-alpha antibody with the interleukin-1 receptor antagonist Eur Cytokine Netw 1995 6: 225–230

    CAS  PubMed  Google Scholar 

  3. MacNaul KL et al. Analysis of IL-1 and TNF-a gene expression in human rheumatoid synoviocytes and normal monocytes by in situ hybridization J Immunol 1990 145: 4154–4166

    CAS  PubMed  Google Scholar 

  4. Elliott MJ et al. Randomized double-blind comparison of chimeric monoclonal antibody to tumor necrosis factor a (cA2) versus placebo in rheumatoid arthritis Lancet 1994 344: 1105–1110

    Article  CAS  Google Scholar 

  5. Moreland LW et al. Treatment of rheumatoid arthritis with a recombinant human tumor necrosis factor receptor (p75)-Fc fusion protein N Engl J Med 1997 337: 141–147

    Article  CAS  Google Scholar 

  6. Hartmann G et al. Specific suppression of human tumor necrosis factor-a synthesis by antisense oligodeoxynucleotides Antisense Nucleic Acid Drug Dev 1996 6: 291–299

    Article  CAS  Google Scholar 

  7. Mayne M, Ni W, McKenna R, Power C . Antisense oligodeoxynucleotides targeting internal exon sequences efficiently regulate TNF-α expression Antisense Nucleic Acid Drug Dev 1999 9: 135–144

    Article  CAS  Google Scholar 

  8. Taylor M, Weller D, Zobzik L . Effect of TNF-α antisense oligomers on cytokine production by primary alveolar macrophages Antisense Nucleic Acid Drug Dev 1998 8: 199–205

    Article  CAS  Google Scholar 

  9. Tu G-C, Cao Q, Zhou F, Israel Y . Tetranucleotide GGGA motif in primary RNA transcripts J Immunol 1998 273: 25125–25131

    CAS  Google Scholar 

  10. d'Hellencourt C, Diaw L, Cornillet P, Guenounou M . Inhibition of human TNFa and LT in cell-free extracts and in cell culture by antisense oligonucleotides Biochim Biophys Acta 1996 1317: 168–174

    Article  Google Scholar 

  11. Evans CH et al. Gene therapy for rheumatic diseases Arthritis Rheum 1999 42: 1–16

    Article  CAS  Google Scholar 

  12. Miller N, Whelan J . Progress in transcriptionally targeted and regulatable vectors for genetic therapy Hum Gene Ther 1997 8: 803–815

    Article  CAS  Google Scholar 

  13. Small JA et al. The visna virus long terminal repeat directs expression of a reporter gene in activated macrophages, lymphocytes, and the central nervous systems of transgenic mice J Virol 1989 63: 1891–1896

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Bellosta S et al. Macrophage-specific expression of human apolipoprotein E reduces atherosclerosis in hypercholesterolemic apolipoprotein E-null mice J Clin Invest 1995 96: 2170–2179

    Article  CAS  Google Scholar 

  15. Deforge LE et al. Biphasic production of IL-8 in lipopolysaccharide (LPS)-stimulated human whole blood: separation of LPS- and cytokine-stimulated components using anti-tumor necrosis factor and anti-IL-1 antibodies J Immunol 1992 148: 2133–2141

    CAS  PubMed  Google Scholar 

  16. Charles P et al. Regulation of cytokines, cytokine inhibitors, and acute-phase proteins following anti-TNF-α therapy in rheumatoid arthritis J Immunol 1999 163: 1521–1528

    CAS  PubMed  Google Scholar 

  17. Zagariya A et al. Tumor necrosis factor alpha gene regulation: enhancement of C/EBPβ-induced activation by c-Jun Mol Cell Biol 1998 18: 2815–2824

    Article  CAS  Google Scholar 

  18. Liu H et al. TNF-α gene expression in macrophages: regulation by NF-κB is independent of c-Jun or C/EBPβ J Immunol 2000 164: 4277–4285

    Article  CAS  Google Scholar 

  19. Pope RM, Leutz A, Ness SA . C/EBPβ regulation of the tumor necrosis factor alpha gene J Clin Invest 1994 94: 1449–1455

    Article  CAS  Google Scholar 

  20. Smits PH, de Wit L, van der Eb AJ, Zantema A . The adenovirus E1A-associated 300 kDa adaptor protein counteracts the inhibition of the collagenase promoter by E1A and represses transformation Oncogene 1996 12: 1529–1535

    CAS  Google Scholar 

  21. Clarke S et al. The human lysozyme promoter directs reporter gene expression to activated myelomonocytic cells in transgenic mice Proc Natl Acad Sci USA 1996 93: 1434–1438

    Article  CAS  Google Scholar 

  22. Economou JS et al. Genetic analysis of the human tumor necrosis factor α/cachectin promoter region in a macrophage cell line J Exp Med 1989 170: 321–326

    Article  CAS  Google Scholar 

  23. Pope R, Mungre S, Liu H, Thimmapaya B . Regulation of TNF-a expression in normal macrophages: role of C/EBPβ Cytokine 2000 12: 1171–1181

    Article  CAS  Google Scholar 

  24. Ghadge GD, Swaminathan S, Katze M, Thimmapaya B . Binding of the adenovirus VAI RNA to interferon induced 68 kDa protein kinase correlates with function Proc Natl Acad Sci USA 1991 88: 7140–7145

    Article  CAS  Google Scholar 

  25. Thimmappaya B, Weinberger C, Schneider RJ, Shenk T . Adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection Cell 1982 31: 543–551

    Article  CAS  Google Scholar 

  26. Kaspers GJ et al. In vitro cellular drug resistance and prognosis in newly diagnosed childhood acute lymphoblastic leukemia Blood 1997 90: 2723–2729

    CAS  Google Scholar 

  27. Coll J et al. Mapping by in vitro constructs of the P100gag-mil region, accounting for induction of chicken neuroretina cell proliferation J Virol 1988 62: 2808–2816

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work supported by NO1-AR-6–2229, from the National Institute of Arthritis, Musculskeletal and Skin Diseases. We thank Harris Perlman, PhD for helpful discussions and for assistance in preparation of the Figures.

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Authors and Affiliations

  1. Department of Medicine, Division of Rheumatology, Lakeside Division, Northwestern University Medical School and the VA Chicago Health Care System, Chicago, IL, USA

    P Sidiropoulos, H Liu, S Mungre & RM Pope

  2. Department of Microbiology and Immunology, Northwestern University Medical School, Chicago, IL, USA

    L Anderson & B Thimmapaya

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  1. P Sidiropoulos
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  2. H Liu
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Sidiropoulos, P., Liu, H., Mungre, S. et al. Efficacy of adenoviral TNFα antisense is enhanced by a macrophage specific promoter. Gene Ther 8, 223–231 (2001). https://doi.org/10.1038/sj.gt.3301368

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