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.

  • Research Article
  • Published:

Retroinfusion of NFκB decoy oligonucleotide extends cardioprotection achieved by CD18 inhibition in a preclinical study of myocardial ischemia and retroinfusion in pigs

Gene Therapy volume 9, pages 518–526 (2002)Cite this article

Abstract

Myocardial reperfusion injury is partially mediated by postischemic inflammation. Beyond acute PMN recruitment, postischemic inflammation comprises subacute PMN adhesion, eg via NFκB activation. In a pig model of 60-min LAD occlusion by PTCA ballon inflation and 1 to 7 days of reperfusion, we investigated the impact of targeted NFκB decoy oligonucleotide (ODN) transfection in the area at risk (AAR) on infarct size and regional myocardial function. After 55 min of LAD occlusion, liposomes containing NFκB ODN were selectively retroinfused into the anterior interventricular vein for 5 min. Then, retroinfusion was stopped and reperfusion was initiated. Where indicated, CD18 antibody IB4 was infused systemically at 30 min of ischemia. Methylen blue and tetrazolium-red staining were used for quantification of the infarct size. Subendocardial segment shortening (SES) by sonomicrometric crystals in infarct area and AAR was assessed under pacing (expressed as % of control region). NFκB decoy ODN retroinfusion reduced infarct size (36 ± 4% versus 49 ± 5% in control hearts at day 7), whereas functional reserve of the AAR (SES 73 ± 17% versus 46 ± 18% at 180/min) tended to improve. Similar effects were observed after IB4 infusion (38 ± 5% infarct size, 85 ± 7% SES at 180/min). A combination of NFκB decoy ODN retroinfusion and IB4 infusion further decreased infarct size (26 ± 2%) and improved functional reserve (SES 94 ± 6% at 180/min). We conclude that NFκB decoy ODN transfection by retroinfusion is feasible in pig hearts and provides postischemic cardioprotection in addition to CD18 blockade.

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

Similar content being viewed by others

References

  1. Duilio C. et al. Neutrophils are primary source of O2 radicals during reperfusion after prolonged myocardial ischemia Am J Physiol 2001 280: H2649 H2649

    CAS  Google Scholar 

  2. Lefer D.J. et al. Cardioprotective actions of a monoclonal antibody against CD-18 in myocardial ischemia-reperfusion injury Circulation 1993 88: 1779 1779

    Article  CAS  Google Scholar 

  3. Paroczai M. et al. Effects of bisaramil on coronary-occlusion-reperfusion injury and free-radical-induced reactions Pharmacol Res 1996 33: 327 327

    Article  CAS  Google Scholar 

  4. Simpson P.J. et al. Reduction of experimental canine myocardial reperfusion injury by a monoclonal antibody (anti-Mo1, anti-CD11b) that inhibits leukocyte adhesion J Clin Invest 1988 81: 624 624

    Article  CAS  Google Scholar 

  5. Tanaka M. et al. Effect of anti-CD18 antibody on myocardial neutrophil accumulation and infarct size after ischemia and reperfusion in dogs Circulation 1993 87: 526 526

    Article  CAS  Google Scholar 

  6. Flynn D.M., Buda A.J., Jeffords P.R., Lefer D.J. . A sialyl Lewis(x)-containing carbohydrate reduces infarct size: role of selectins in myocardial reperfusion injury Am J Physiol 1996 271: H2086 H2086

    CAS  PubMed  Google Scholar 

  7. Simpson P.J. et al. Sustained limitation of myocardial reperfusion injury by a monoclonal antibody that alters leukocyte function Circulation 1990 81: 226 226

    Article  CAS  Google Scholar 

  8. Barnes P.J., Karin M. . Nuclear factor-kappaB: a pivotal transcription factor in chronic inflammatory diseases New Engl J Med 1997 336: 1066 1066

    Article  CAS  Google Scholar 

  9. Kupatt C. et al. TNF alpha contributes to ischemia/reperfusion induced endothelial activation in isolated hearts Circ Res 1999 84: 392 392

    Article  CAS  Google Scholar 

  10. Chandrasekar B., Freeman G.I. . Induction of nuclear factor kappaB and activation protein 1 in postischemic myocardium FEBS Lett 1997 401: 30 30

    Article  CAS  Google Scholar 

  11. Morishita R. et al. In vivo transfection of cis element ‘decoy’ against nuclear factor-kappaB binding site prevents myocardial infarction Nature Med 1997 3: 894 894

    Article  CAS  Google Scholar 

  12. Morishita R., Higaki J., Tomita N., Ogihara T. . Application of transcription factor ‘decoy’ strategy as means of gene therapy and study of gene expression in cardiovascular disease Circ Res 1998 82: 1023 1023

    Article  CAS  Google Scholar 

  13. Kokura S. et al. Molecular mechanisms of neutrophil – endothelial cell adhesion induced by redox imbalance Circ Res 1999 84: 516 516

    Article  CAS  Google Scholar 

  14. Ichikawa H. et al. Molecular mechanisms of anoxia/reoxygenation-induced neutrophil adherence to cultured endothelial cells Circ Res 1997 81: 922 922

    Article  CAS  Google Scholar 

  15. Dove A. . CD18 trials disappoint again Nature Biotechnol 2000 18: 817 817

    Article  CAS  Google Scholar 

  16. Baran K.W. et al. Double-blind, randomized trial of an anti-CD18 antibody in conjunction with recombinant tissue plasminogen activator for acute myocardial infarction Circulation 2001 104: 2778 2778

    Article  CAS  Google Scholar 

  17. Boekstegers P. et al. Myocardial gene transfer by selective pressure-regulated retroinfusion of coronary veins Gene Therapy 2000 7: 232 232

    Article  CAS  Google Scholar 

  18. Bergmann M.W., Loser P., Dietz R., Harsdorf R. . Effect of NF-kappa B Inhibition on TNF-alpha-induced apoptosis and downstream pathways in cardiomyocytes J Mol Cell Cardiol 2001 33: 1223 1223

    Article  CAS  Google Scholar 

  19. Geng J.G. et al. Rapid neutrophil adhesion to activated endothelium mediated by GMP-140 Nature 1990 343: 757 757

    Article  CAS  Google Scholar 

  20. Milhoan K.A., Lane T.A., Bloor C.M. . Hypoxia induces endothelial cells to increase their adherence for neutrophils: role of PAF Am J Physiol 1992 263: H956 H956

    CAS  PubMed  Google Scholar 

  21. Jones S.P. et al. Leukocyte and endothelial cell adhesion molecules in a chronic murine model of myocardial reperfusion injury Am J Physiol 2000 279: H2196 H2196

    CAS  Google Scholar 

  22. Kukielka G.L. et al. Induction of interleukin-6 synthesis in the myocardium. Potential role in postreperfusion inflammatory injury Circulation 1995 92: 1866 1866

    Article  CAS  Google Scholar 

  23. Kumar A.G. et al. Induction of monocyte chemoattractant protein-1 in the small veins of the ischemic and reperfused canine myocardium Circulation 1997 95: 693 693

    Article  CAS  Google Scholar 

  24. Kupatt C. et al. ACE-inhibition prevents postischemic coronary leukocyte adhesion and leukocyte-dependent reperfusion injury Cardiovasc Res 1997 36: 386 386

    Article  CAS  Google Scholar 

  25. Jeremias I. et al. Involvement of CD95/Apo-1/Fas in apoptotic cell death following myocardial infarction Circulation 2000 102: 915 915

    Article  CAS  Google Scholar 

  26. Bauer M.K.A. et al. Role of reactive oxygen intermediates in activation-induced CD95 (APO- 1/Fas) ligand expression J Biol Chem 1998 273: 8048 8048

    Article  CAS  Google Scholar 

  27. Sanlioglu S. et al. Lipopolysaccharide induces Racl-dependent reactive oxygen species formation and coordinates tumor necrosis factor-alpha secretion through IKK regulation of NF-kappa B J Biol Chem 2001 276: 30188 30188

    Article  CAS  Google Scholar 

  28. Rivera-Walsh I. et al. NF-kappa B signaling pathway governs TRAIL gene expression and human T cell leukemia virus-1 tax-induced T cell death J Biol Chem 2001 276: 40385 40385

    Article  CAS  Google Scholar 

  29. Hammerman H. et al. Dose-dependent effects of short-term methylprednisolone on myocardial infarct extent, scar formation and ventricular function Circulation 1983 68: 446 446

    Article  CAS  Google Scholar 

  30. Roberts R., DeMello V., Sobel B.E. . Deleterious effects of methylprednisolone in patients with myocardial infarction Circulation 1976 53: 204 204

    Article  Google Scholar 

  31. Heymans S. et al. Inhibition of plasminogen activators or matrix metalloproteinases prevents cardiac rupture but impairs therapeutic angiogenesis and causes cardiac failure Nat Med 1999 5: 1135 1135

    Article  CAS  Google Scholar 

  32. Ducharme A. et al. Targeted deletion of matrix metalloproteinase-9 attenuates left ventricular enlargement and collagen accumulation after experimental myocardial infarction J Clin Invest 2000 106: 55 55

    Article  CAS  Google Scholar 

  33. Bond M., Fabunmi R.P., Baker A.H., Newby A.C. . Synergistic upregulation of metalloproteinase-9 by growth factors and inflammatory cytokines: an absolute requirement for transcription factor NF-kappa B FEBS Lett 1998 435: 29 29

    Article  CAS  Google Scholar 

  34. Hansen S.K. et al. A novel complex between the p65 subunit of NF-kappa B and c-Rel binds to a DNA element involved in the phorbol ester induction of the human urokinase gene EMBO J 1992 11: 205 205

    Article  CAS  Google Scholar 

  35. Boekstegers P., Giehrl W., Degenfeld Gv., Steinbeck G. . Selective suction and pressure-regulated retroinfusion; an effective and safe approach to retrograde protection against myocardial ischemia in patients undergoing normal and high risk percutaneous transluminal angioplasty J Am Coll Cardiol 1998 31: 1525 1525

    Article  CAS  Google Scholar 

  36. Nishida M. et al. Isolation and characterization of human and rat cardiac microvascular endothelial cells Am J Physiol 1993 264: 639 639

    Google Scholar 

  37. Jahnke A., Johnson J.P. . Synergistic activation of intercellular adhesion molecule 1 (ICAM-1) by TNF-alpha and IFN-gamma is mediated by p65/p50 and p65/c-Rel and interferon-responsive factor Stat1 alpha (p91) that can be activated by both IFN-gamma and IFN-alpha FEBS Lett 1994 354: 220 220

    Article  CAS  Google Scholar 

  38. Degenfeld Gv., Giehrl W., Boekstegers P. . Targeting of dobutamine to ischemic myocardium without systemic effects by selective suction and pressure-regulated retroinfusion Cardiovasc Res 1997 35: 233 233

    Article  Google Scholar 

  39. Boekstegers P. et al. Preservation of regional myocardial function and myocardial oxygen tension during acute ischemia in pigs: comparison of selective synchronized suction and retroinfusion of coronary veins to synchronized coronary venous retroperfusion J Am Coll Cardiol 1994 23: 459 459

    Article  CAS  Google Scholar 

  40. Boekstegers P., Diebold J., Weiss C. . Selective ECG synchronised suction and retroinfusion of coronary veins: first results of studies in acute myocardial ischemia in dogs Cardiovasc Res 1990 24: 456 456

    Article  CAS  Google Scholar 

  41. Zahler S., Kupatt C., Becker B.F. . Endothelial preconditioning by transient oxidative stress reduces inflammatory responses of cultured endothelial cells to TNF-alpha FASEB J 2000 14: 555 555

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The expert assistance of Susanne Helbig and Elisabeth Ronft is gratefully acknowledged. This work has been supported by the Deutsche Forschungsgemeinschaft (for 440/1–1). We wish to thank Konrad Messmer, Director of the Institute for Surgical Research, for the opportunity to conduct all animal experiments in this institution, and Helmut Habazettl, as well as Fritz Krombach for helpful advice.

Author information

Authors and Affiliations

  1. Internal Medicine I, Klinikum Großhadern, Munich, Germany

    C Kupatt, R Wichels, M Deiß, A Molnar, C Lebherz, P Raake, G von Degenfeld, D Hahnel & P Boekstegers

Authors
  1. C Kupatt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R Wichels
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Deiß
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Molnar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C Lebherz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. P Raake
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. G von Degenfeld
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. D Hahnel
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. P Boekstegers
    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

Kupatt, C., Wichels, R., Deiß, M. et al. Retroinfusion of NFκB decoy oligonucleotide extends cardioprotection achieved by CD18 inhibition in a preclinical study of myocardial ischemia and retroinfusion in pigs. Gene Ther 9, 518–526 (2002). https://doi.org/10.1038/sj.gt.3301673

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301673

Keywords

This article is cited by

Search

Advanced search

Quick links