Ex vivo gene therapy prevents chronic graft vascular disease in cardiac allografts

J Thorac Cardiovasc Surg. 1998 Sep;116(3):386-96. doi: 10.1016/s0022-5223(98)70003-0.

Abstract

Objective: We hypothesized that ex vivo hyperbaric transfection of antisense oligodeoxynucleotides for blockade of intercellular adhesion molecule-1, an important mediator of cell adhesion and T-cell co-stimulation, would reduce chronic graft vascular disease in cardiac allografts.

Methods: PVG hearts underwent ex vivo transfection with antisense, reverse antisense intercellular adhesion molecule-1 oligodeoxynucleotide (80 micromol/L), or saline solution at 3 atm pressure for 45 minutes at 4 degrees C and were transplanted heterotopically into ACI recipients with or without treatment with intercellular adhesion molecule-1 (1A29) or leukocyte function associated antigen-1 (WT.1) monoclonal antibodies. Transfection efficiency was confirmed with fluorescein isothiocyanate-labeled oligodeoxynucleotides and fluorescent microscopy. Efficacy of intracellular adhesion molecule-1 blockade was assessed with the use of immunohistochemistry. Graft reperfusion injury was evaluated at 6 to 24 hours by neutrophil infiltration (myeloperoxidase [MPO]), cardiac edema (%wt/wt), and histologic injury (percent contraction band necrosis). Grafts from recipients treated with cyclosporine A (5 mg/kg per day, days 0 to 9) were scored for chronic graft vascular disease on postoperative day 90 ranging from 0 (no involvement) to 4 (>50% vascular occlusion).

Results: Transfection was highly efficient (fluorescein isothiocyanate-labeled oligodeoxynucleotides in 48%+/-5% of total myocardial nuclei) and effective at blocking intracellular adhesion molecule-1 expression (positive area in allografts taken on postoperative day 3 was reduced from 100%+/-0% to 52%+/-14%, n=4). Blockade with antisense oligodeoxynucleotides versus monoclonal antibodies was less effective at preventing reperfusion injury while more effective at reducing chronic graft vascular disease (score 0.98+/-0.48, p < 0.05). Reverse antisense oligodeoxynucleotides and vector control (antisense oligodeoxynucleotide infusion without pressure) groups failed to demonstrate this beneficial effect.

Conclusion: Hyperbaric transfection of antisense oligodeoxynucleotides proved highly efficient, effective at blockade of intracellular adhesion molecule-1, and demonstrated a sequence-specific reduction in chronic graft vascular disease. This highly targeted alteration of donor organ immunogenicity may have an important future role in clinical immunosuppressive strategies.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Coronary Disease / etiology
  • Coronary Disease / immunology
  • Coronary Disease / prevention & control*
  • Genetic Therapy / methods*
  • Graft Rejection / immunology
  • Graft Rejection / prevention & control
  • Heart Transplantation* / adverse effects
  • Immunosuppression / methods
  • Intercellular Adhesion Molecule-1 / genetics*
  • Intercellular Adhesion Molecule-1 / physiology
  • Male
  • Myocardial Reperfusion Injury / etiology
  • Myocardial Reperfusion Injury / immunology
  • Myocardial Reperfusion Injury / prevention & control*
  • Oligonucleotides, Antisense / therapeutic use
  • Postoperative Complications / etiology
  • Postoperative Complications / immunology
  • Postoperative Complications / prevention & control*
  • Pressure
  • Rats
  • Rats, Inbred ACI
  • Rats, Inbred Strains
  • Transfection*
  • Transplantation, Heterotopic

Substances

  • Oligonucleotides, Antisense
  • Intercellular Adhesion Molecule-1