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Published on: August 12, 2014
In vivo adenovirus-mediated endothelial nitric oxide synthase gene transfer ameliorates lung allograft
1Division of Cardiothoracic Surgery, the Department of Surgery, Washington University School of Medicine, Barnes-Jewish Hospital, St Louis, MO 63110, USA.
Objective:
Nitric oxide regulates vascular tone, inhibits platelet aggregation, and inhibits leukocyte adhesion, all of which are important modulators of ischemia-reperfusion injury. This study aimed to determine the effects of endothelial constitutive nitric oxide synthase gene transfer on ischemia-reperfusion injury in a rat lung transplant model.
Methods:
In group I, donor animals were injected intravenously with 5 x 10(9) pfu of adenovirus-encoding endothelial constitutive nitric oxide synthase. Groups II and III served as controls, whereby donor animals were injected with either 5 x 10(9) pfu of adenovirus encoding beta-galactosidase or saline solution, respectively. Twenty-four hours after injection, left lungs were harvested and preserved for 18 hours at 4 degrees C, then implanted into isogeneic recipients, which were put to death 24 hours later. Recombinant endothelial constitutive nitric oxide synthase gene expression was evaluated by Western blotting and immunohistochemistry. Lung grafts were assessed by measuring arterial oxygenation, myeloperoxidase activity, and wet/dry weight ratios.
Results:
Western blotting confirmed the overexpression of endothelial constitutive nitric oxide synthase in lungs so transfected compared with controls. Twenty-four hours after reperfusion, mean arterial oxygenation was significantly improved in group I compared with group II and III controls (189.4 +/- 47.1 mm Hg vs 71.7 +/- 8.9 mm Hg and 67.8 +/- 12.2 mm Hg, P =.02, P =.01, respectively). Myeloperoxidase activity, a reflection of tissue neutrophil sequestration, was also significantly reduced in group I compared with groups II and III (0.136 +/- 0.038 DeltaOD/mg/min vs 0. 587 +/- 0.077 and 0.489 +/- 0.126 DeltaOD/mg/min, P =.001, P =.01, respectively).
Conclusion:
Adenovirus-mediated gene transfer with endothelial constitutive nitric oxide synthase ameliorates ischemia-reperfusion injury as manifested by significantly improved oxygenation and decreased neutrophil sequestration in transplanted lung isografts. Endothelial constitutive nitric oxide synthase gene transfer may reduce acute lung dysfunction after lung transplantation.
Insights
Gene transfer of endothelial nitric oxide synthase improved lung transplant outcomes by enhancing oxygenation and reducing neutrophil buildup. This approach shows promise for mitigating ischemia-reperfusion injury in lung grafts.
Area of Science:
- Cardiovascular Biology
- Transplantation Immunology
- Gene Therapy
Background:
- Nitric oxide (NO) is crucial for regulating vascular tone, platelet aggregation, and leukocyte adhesion, all key factors in ischemia-reperfusion (I/R) injury.
- I/R injury significantly impacts the success of organ transplantation, particularly in lung grafts.
Purpose of the Study:
- To investigate the therapeutic potential of endothelial constitutive nitric oxide synthase (ecNOS) gene transfer in mitigating I/R injury in a rat lung transplant model.
- To assess the impact of ecNOS gene transfer on graft function and inflammatory markers post-transplantation.
Main Methods:
- Donor rats received intravenous adenovirus encoding ecNOS (Group I) or a control adenovirus/saline (Groups II/III).
- Left lungs were harvested, preserved, and transplanted into isogeneic recipients.
- Graft function was evaluated by arterial oxygenation, myeloperoxidase activity (neutrophil infiltration), and wet/dry weight ratios 24 hours post-reperfusion.
Main Results:
- Western blotting confirmed successful ecNOS overexpression in Group I lungs.
- Group I demonstrated significantly improved arterial oxygenation compared to controls (189.4 mmHg vs. 71.7-67.8 mmHg).
- Myeloperoxidase activity was significantly reduced in Group I, indicating decreased neutrophil sequestration.
Conclusions:
- Adenovirus-mediated ecNOS gene transfer effectively ameliorates I/R injury in transplanted rat lungs.
- This gene therapy approach significantly improves graft oxygenation and reduces inflammatory cell infiltration.
- ecNOS gene transfer holds promise for reducing acute lung dysfunction following lung transplantation.
