Recombinant endothelial nitric oxide synthase-transduced human saphenous veins: Gene therapy to augment nitric oxide production in bypass conduits

David G. Cable, Timothy O'Brien, Hartzell V Schaff, Vincent J. Pompili

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Background: Nitric oxide is a potent vasodilator that also inhibits platelet aggregation and smooth muscle cell proliferation, properties that may prevent early and late occlusion of saphenous vein coronary bypass conduits. We determined whether human saphenous veins can be transduced with adenovirus vector-encoding bovine endothelial nitric oxide synthase (Ad. CMVeNOS), resulting in functional expression of recombinant nitric oxide synthase. Methods and Results: Harvested segments of human saphenous vein were exposed for 1 hour at 37°C to replication-deficient Ad.CMVeNOS (5x109 PFU/mL) or control adenovirus-encoding Escherichia coli β-galactosidase (Ad.CMVLacZ; 5x109 PFU/mL). The vein segments were analyzed for recombinant endothelial nitric oxide synthase expression and activity 48 hours later. Histochemical staining for recombinant β-galactosidase activity was localized to the luminal endothelium and adventitia of vein segments transduced with Ad.CMVLacZ. Similarly, immunohistochemical staining with a monoclonal antibody for nitric oxide synthase localized recombinant gene expression to endothelial and adventitial cells in Ad. CMVeNOS veins; only endogenous nitric oxide synthase was identified in the endothelium of Ad.CMVLacZ veins. Nitrite generation after stimulation with calcium ionophore increased in Ad.CMVeNOS veins (1420.0±298.2 nM/mg versus 130.3±19.9 nM/mg; n=3; P<.05). Isometric tension recording demonstrated augmented maximal relaxation to calcium ionophore (32±4.5% versus 17.4±7.4%; n=6; P<.05) after precontraction with norepinephrine. Bioassay superfusion demonstrated a twofold augmentation of the biodetector ring relaxation during calcium ionophore stimulation of Ad.CMVeNOS veins. Conclusions: Adenovirus-mediated gene transfer to human saphenous veins resulted in functional transgene expression with increased nitric oxide release. These or similar molecular techniques to increase nitric oxide production may reduce the risk of early thrombosis in saphenous vein grafts.

Original languageEnglish (US)
JournalCirculation
Volume96
Issue number9 SUPPL.
StatePublished - Nov 4 1997

Fingerprint

Nitric Oxide Synthase Type III
Saphenous Vein
Genetic Therapy
Veins
Nitric Oxide
Calcium Ionophores
Adenoviridae
Galactosidases
Nitric Oxide Synthase
Adventitia
Endothelium
Staining and Labeling
Nitrites
Vasodilator Agents
Transgenes
Platelet Aggregation
Biological Assay
Smooth Muscle Myocytes
Norepinephrine
Thrombosis

Keywords

  • Adenoviral vector
  • Gene therapy
  • Nitric oxide synthase
  • Organ chamber
  • Saphenous vein

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Recombinant endothelial nitric oxide synthase-transduced human saphenous veins : Gene therapy to augment nitric oxide production in bypass conduits. / Cable, David G.; O'Brien, Timothy; Schaff, Hartzell V; Pompili, Vincent J.

In: Circulation, Vol. 96, No. 9 SUPPL., 04.11.1997.

Research output: Contribution to journalArticle

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title = "Recombinant endothelial nitric oxide synthase-transduced human saphenous veins: Gene therapy to augment nitric oxide production in bypass conduits",
abstract = "Background: Nitric oxide is a potent vasodilator that also inhibits platelet aggregation and smooth muscle cell proliferation, properties that may prevent early and late occlusion of saphenous vein coronary bypass conduits. We determined whether human saphenous veins can be transduced with adenovirus vector-encoding bovine endothelial nitric oxide synthase (Ad. CMVeNOS), resulting in functional expression of recombinant nitric oxide synthase. Methods and Results: Harvested segments of human saphenous vein were exposed for 1 hour at 37°C to replication-deficient Ad.CMVeNOS (5x109 PFU/mL) or control adenovirus-encoding Escherichia coli β-galactosidase (Ad.CMVLacZ; 5x109 PFU/mL). The vein segments were analyzed for recombinant endothelial nitric oxide synthase expression and activity 48 hours later. Histochemical staining for recombinant β-galactosidase activity was localized to the luminal endothelium and adventitia of vein segments transduced with Ad.CMVLacZ. Similarly, immunohistochemical staining with a monoclonal antibody for nitric oxide synthase localized recombinant gene expression to endothelial and adventitial cells in Ad. CMVeNOS veins; only endogenous nitric oxide synthase was identified in the endothelium of Ad.CMVLacZ veins. Nitrite generation after stimulation with calcium ionophore increased in Ad.CMVeNOS veins (1420.0±298.2 nM/mg versus 130.3±19.9 nM/mg; n=3; P<.05). Isometric tension recording demonstrated augmented maximal relaxation to calcium ionophore (32±4.5{\%} versus 17.4±7.4{\%}; n=6; P<.05) after precontraction with norepinephrine. Bioassay superfusion demonstrated a twofold augmentation of the biodetector ring relaxation during calcium ionophore stimulation of Ad.CMVeNOS veins. Conclusions: Adenovirus-mediated gene transfer to human saphenous veins resulted in functional transgene expression with increased nitric oxide release. These or similar molecular techniques to increase nitric oxide production may reduce the risk of early thrombosis in saphenous vein grafts.",
keywords = "Adenoviral vector, Gene therapy, Nitric oxide synthase, Organ chamber, Saphenous vein",
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T1 - Recombinant endothelial nitric oxide synthase-transduced human saphenous veins

T2 - Gene therapy to augment nitric oxide production in bypass conduits

AU - Cable, David G.

AU - O'Brien, Timothy

AU - Schaff, Hartzell V

AU - Pompili, Vincent J.

PY - 1997/11/4

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N2 - Background: Nitric oxide is a potent vasodilator that also inhibits platelet aggregation and smooth muscle cell proliferation, properties that may prevent early and late occlusion of saphenous vein coronary bypass conduits. We determined whether human saphenous veins can be transduced with adenovirus vector-encoding bovine endothelial nitric oxide synthase (Ad. CMVeNOS), resulting in functional expression of recombinant nitric oxide synthase. Methods and Results: Harvested segments of human saphenous vein were exposed for 1 hour at 37°C to replication-deficient Ad.CMVeNOS (5x109 PFU/mL) or control adenovirus-encoding Escherichia coli β-galactosidase (Ad.CMVLacZ; 5x109 PFU/mL). The vein segments were analyzed for recombinant endothelial nitric oxide synthase expression and activity 48 hours later. Histochemical staining for recombinant β-galactosidase activity was localized to the luminal endothelium and adventitia of vein segments transduced with Ad.CMVLacZ. Similarly, immunohistochemical staining with a monoclonal antibody for nitric oxide synthase localized recombinant gene expression to endothelial and adventitial cells in Ad. CMVeNOS veins; only endogenous nitric oxide synthase was identified in the endothelium of Ad.CMVLacZ veins. Nitrite generation after stimulation with calcium ionophore increased in Ad.CMVeNOS veins (1420.0±298.2 nM/mg versus 130.3±19.9 nM/mg; n=3; P<.05). Isometric tension recording demonstrated augmented maximal relaxation to calcium ionophore (32±4.5% versus 17.4±7.4%; n=6; P<.05) after precontraction with norepinephrine. Bioassay superfusion demonstrated a twofold augmentation of the biodetector ring relaxation during calcium ionophore stimulation of Ad.CMVeNOS veins. Conclusions: Adenovirus-mediated gene transfer to human saphenous veins resulted in functional transgene expression with increased nitric oxide release. These or similar molecular techniques to increase nitric oxide production may reduce the risk of early thrombosis in saphenous vein grafts.

AB - Background: Nitric oxide is a potent vasodilator that also inhibits platelet aggregation and smooth muscle cell proliferation, properties that may prevent early and late occlusion of saphenous vein coronary bypass conduits. We determined whether human saphenous veins can be transduced with adenovirus vector-encoding bovine endothelial nitric oxide synthase (Ad. CMVeNOS), resulting in functional expression of recombinant nitric oxide synthase. Methods and Results: Harvested segments of human saphenous vein were exposed for 1 hour at 37°C to replication-deficient Ad.CMVeNOS (5x109 PFU/mL) or control adenovirus-encoding Escherichia coli β-galactosidase (Ad.CMVLacZ; 5x109 PFU/mL). The vein segments were analyzed for recombinant endothelial nitric oxide synthase expression and activity 48 hours later. Histochemical staining for recombinant β-galactosidase activity was localized to the luminal endothelium and adventitia of vein segments transduced with Ad.CMVLacZ. Similarly, immunohistochemical staining with a monoclonal antibody for nitric oxide synthase localized recombinant gene expression to endothelial and adventitial cells in Ad. CMVeNOS veins; only endogenous nitric oxide synthase was identified in the endothelium of Ad.CMVLacZ veins. Nitrite generation after stimulation with calcium ionophore increased in Ad.CMVeNOS veins (1420.0±298.2 nM/mg versus 130.3±19.9 nM/mg; n=3; P<.05). Isometric tension recording demonstrated augmented maximal relaxation to calcium ionophore (32±4.5% versus 17.4±7.4%; n=6; P<.05) after precontraction with norepinephrine. Bioassay superfusion demonstrated a twofold augmentation of the biodetector ring relaxation during calcium ionophore stimulation of Ad.CMVeNOS veins. Conclusions: Adenovirus-mediated gene transfer to human saphenous veins resulted in functional transgene expression with increased nitric oxide release. These or similar molecular techniques to increase nitric oxide production may reduce the risk of early thrombosis in saphenous vein grafts.

KW - Adenoviral vector

KW - Gene therapy

KW - Nitric oxide synthase

KW - Organ chamber

KW - Saphenous vein

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