Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Nonviral Transfer Technology
  • Published:

Transcription factor decoy for NFκB inhibits TNF-α-induced cytokine and adhesion molecule expression in vivo

Gene Therapy volume 7, pages 1326–1332 (2000)Cite this article

Abstract

The expression of several cytokines and adhesion molecules is regulated by the transcription factor NFκB, which is activated by tumor necrosis factor alpha (TNF-α). In this study, we employed a mouse model of nephritis induced by TNF-α to examine whether inhibition of NFκB activity using transcription factor decoy oligonucleotides (ODN) blocks cytokine and adhesion molecule expression and attenuates the renal inflammatory response. First, we developed a method for delivering FITC-ODN in vivo into mouse kidney glomeruli by employing HVJ-liposome. Then, in order to study the feasibility of decoy strategy in vivo, the reporter gene chloramphenicol acetyltransferase (CAT) driven by three tandemly repeated NFκB binding sequences was transfected into the kidney. Intrapenetorial injection of TNF-α stimulated CAT expression in vivo, and the increase in CAT expression was completely abolished by NFκB decoy ODN, but not scrambled ODN. Therefore, we examined the effect of NFκB decoy ODN transfection on TNF-α-induced endogenous interleukin (IL)-1α, IL-1β, IL-6, ICAM-1 and VCAM-1 gene expression as assessed by RT-PCR and Northern blotting. Our present data showed that NFκB decoy, but not scrambled, ODN abolished TNF-α induced gene expression in vivo, as well as glomerular inflammation as assessed by CD45 staining. Taken together, our results suggest the potential utility of NFκB decoy strategy for molecular therapy to glomerular inflammatory diseases.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Egido J et al. Role of tumor necrosis factor-alpha in the pathogenesis of glomerular diseases Kidney Int 1993 39: S59–S64

    CAS  Google Scholar 

  2. Sedor JR et al. Cytokines, mesangial cell activation and glomerular injury Kidney Int 1993 39: S65–S70

    CAS  Google Scholar 

  3. Border WA et al. Suppression of experimental glomerulonephritis by antiserum against transforming growth factor beta 1 Nature 1990 346: 371–374

    Article  CAS  Google Scholar 

  4. Gesualdo L et al. Platelet-derived growth factor and proliferative glomerulonephritis Kidney Int 1993 39: S86–S89

    CAS  Google Scholar 

  5. Chow J, Hartley RB, Jagger C, Dilly SA . ICAM-1 expression in renal disease J Clin Pathol 1992 45: 880–884

    Article  CAS  Google Scholar 

  6. Floege J, Eng E, Young BA, Johnson R . Factors involved in the regulation of mesangial cell proliferation in vitro and in vivo Kidney Int 1993 39: S47–S54

    CAS  Google Scholar 

  7. Sterzel RB, Schulze-Lohoff E, Marx M . Cytokines and mesangial cells Kidney Int 1993 39: S26–S31

    CAS  Google Scholar 

  8. Lenardo MJ, Baltimore D . NF-kappa B: a pleitropic mediator of inducible and tissue specific gene control Cell 1989 58: 227–229

    Article  CAS  Google Scholar 

  9. Libermann TA, Baltimore D . Activation of interleukin-6 gene expression through the NF-kappa B transcription factor Mol Cell Biol 1990 10: 2327–2334

    Article  CAS  Google Scholar 

  10. Satriano J, Schlondorff D . Activation of attenuattion of transcriptional factor NF-κB in mouse glomerular cells in response to tumor necrosis factor-, immunoglobulin G, and adenosis 3′-5′-cyclic monophosphate J Clin Invest 1994 94: 1629–1636

    Article  CAS  Google Scholar 

  11. Neish AS et al. Functional analysis of the human vascular cell adhesion molecule 1 promoter J Exp Med 1992 176: 1583–1593

    Article  CAS  Google Scholar 

  12. Brennan DC, Jevnikar AM, Takei F, Reubin-Kelley VE . Mesangial cell accessory functions: mediation by intracellular adhesion molecule-1 Kidney Int 1990 38: 1039–1046

    Article  CAS  Google Scholar 

  13. Bielinska A, Shivdasani RA, Zhang L, Nabel GJ . Regulation of gene expression with double-stranded phosphorothioate oligonucleotides Science 1990 250: 997–1000

    Article  CAS  Google Scholar 

  14. Sullenger BA, Gallardo HF, Ungers GE, Giboa E . Overexpression of TAR sequence renders cells resistant to human immunodeficiency virus replication Cell 1990 63: 601–608

    Article  CAS  Google Scholar 

  15. Morishita R et al. A novel molecular strategy using cis element ‘decoy’ of E2F binding site inhibits smooth muscle proliferation in vivo Proc Natl Acad Sci USA 1995 92: 5855–5859

    Article  CAS  Google Scholar 

  16. Yamada T et al. In vivo identification of a negative regulatory element in the mouse renin gene using direct gene transfer J Clin Invest 1995 96: 1230–1237

    Article  CAS  Google Scholar 

  17. Tomita N et al. A novel strategy transcription factor decoy for E2F inhibits proliferation in mesangial cells in vitro Am J Physiol 1998 275: F278–F284

    Article  CAS  Google Scholar 

  18. Tomita S et al. Transcription factor decoy to study the molecular mechanism of negative regulation of renin gene expression in the liver in vivo Circ Res 1999 84: 1059–1066

    Article  CAS  Google Scholar 

  19. Tomita N et al. Direct in vivo gene introduction into rat kidney Biochem Biophys Res Commun 1992 186: 129–134

    Article  CAS  Google Scholar 

  20. Isaka Y et al. Glomerulosclerosis induced by in vivo transfection of transforming growth factor-β or platelet derived growth factor into the rat kidney J Clin Invest 1993 92: 2597–2601

    Article  CAS  Google Scholar 

  21. Helin K et al. A cDNA encoding a RB-binding protein with properties of the transcription factor E2F Cell 1992 70: 337–350

    Article  CAS  Google Scholar 

  22. Kaelin WG et al. Expression cloning of a cDNA encoding a retinobalstoma-binding protein with E2F-like properties Cell 1992 70: 351–364

    Article  CAS  Google Scholar 

  23. Woolf AS, Bosch RJ, Fine LG . Gene transfer into the mammalian kidney: first steps towards renal gene therapy Kidney Int 1993 39: S116–S119

    CAS  Google Scholar 

  24. Moullier P et al. Adenoviral-mediated gene transfer to renal tubular cells in vivo Kidney Int 1994 45: 1220–1225

    Article  CAS  Google Scholar 

  25. Morishita R et al. Single intraluminal delivery of antisense cdc 2 kinase and PCNA oligonucleotides results in chronic inhibition of neointimal hyperplasia Proc Natl Acad Sci USA 1993 90: 8474–8479

    Article  CAS  Google Scholar 

  26. Morishita R et al. Intimal hyperplasia after vascular injury is inhibited by antisense cdk 2 kinase oligonucleotides J Clin Invest 1994 93: 1458–1464

    Article  CAS  Google Scholar 

  27. Morishita R et al. Pharmacokinetics of antisense oligonucleotides (cyclin B1 and cdc 2 kinase) in the vessel wall: enhanced therapeutic utility for restenosis by HVJ-liposome method Gene 1994 149: 13–19

    Article  CAS  Google Scholar 

  28. Tomita N et al. Transient decrease in high blood pressure by in vivo transfer of antisense oligodeoxynucleotides against rat angiotensinogen Hypertension 1995 26: 131–136

    Article  CAS  Google Scholar 

  29. Morishita R et al. Systemic administration of HVJ viral coat-liposome complex containing insulin vector decreases glucose level in diabetic mice: model of gene therapy Clin Res 1994 42: 211A (Abstr.)

    Google Scholar 

  30. Morishita R et al. Apolipoprotein E deficient mice created by systemic administration of antisense oligonucleotides: a new model for lipoprotein metabolism study Clin Res 1994 42: 178A (Abstr.)

    Google Scholar 

  31. Iwao M et al. Induction of interleukin 6 synthesis in mouse glomeruli and cultured mesangial cells Nephron 1992 62: 58–65

    Article  Google Scholar 

  32. Horiuchi M et al. Molecular mechanism of tissue-specific regulation of mouse renin gene expression by cAMP J Biol Chem 1991 266: 16247–16254

    CAS  PubMed  Google Scholar 

  33. Horiuchi M, Pratt RE, Nakamura N, Dzau VJ . Distinct nuclear proteins competing for an overlapping sequence of cAMP and negative-regulatory elements regulate tissue-specific mouse renin gene expression J Clin Invest 1993 92: 1805–1811

    Article  CAS  Google Scholar 

  34. Kaneda Y, Iwai K, Uchida T . Increased expression of DNA cointroduced with nuclear protein in adult rat liver Science 1989 243: 375–378

    Article  CAS  Google Scholar 

  35. Kaneda Y, Morishita R, Tomita N . Increased expression of DNA cointroduced with nuclear protein in adult rat liver J Mol Med 1995 73: 289–297

    Article  CAS  Google Scholar 

  36. Dzau VJ, Mann MJ, Morishita R, Kaneda Y . Fusigenic viral liposome for gene therapy in cardiovascular diseases Proc Natl Acad Sci USA 1996 93: 11421–11425

    Article  CAS  Google Scholar 

  37. Saito I et al. Expression of cell adhesion molecules in the salivary and lacrimal glands of Sjogren's syndrome J Clin Lab Anal 1993 7: 180–187

    Article  CAS  Google Scholar 

  38. Araki M, Araki K, Vassalli P . Cloning and sequencing of mouse VCAM-1 cDNA Gene 1993 126: 261–264

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We wish to thank Chihiro Noguchi and Shiori Takase for their excellent technical assistance. This work is supported by NIH grants HL 35610, HL 35252, HL 42663. Dr Gary H Gibbons is the recipient of a Pew Biomedical Scholar Award. Dr Victor J Dzau is the recipient of an NIH MERIT Award HL35610. Dr Ryuichi Morishita is the recipient of a Harry Goldblatt Award from the Council of High Blood Pressure, American Heart Association.

Author information

Authors and Affiliations

  1. Department of Geriatric Medicine, Osaka University Medical School, Japan

    N Tomita, R Morishita, S Tomita, J Higaki & T Ogihara

  2. Department of Medicine, Harvard Medical School, Brigham and Women's Hospital, Boston, MA, USA

    N Tomita, S Tomita, GH Gibbons, L Zhang, M Horiuchi & VJ Dzau

  3. Department of Gene Therapy Science, Osaka University Medical School, Ehime, Japan

    R Morishita & Y Kaneda

  4. Department of Medical Biochemistry, Ehime University School of Medicine, Ehime, Japan

    M Horiuchi

Authors
  1. N Tomita
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R Morishita
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Tomita
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. GH Gibbons
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. L Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M Horiuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Y Kaneda
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. J Higaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. T Ogihara
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. VJ Dzau
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tomita, N., Morishita, R., Tomita, S. et al. Transcription factor decoy for NFκB inhibits TNF-α-induced cytokine and adhesion molecule expression in vivo. Gene Ther 7, 1326–1332 (2000). https://doi.org/10.1038/sj.gt.3301243

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301243

Keywords

This article is cited by

Search

Advanced search

Quick links