Cotransfection of Vascular Endothelial Growth Factor-A and Platelet-Derived Growth Factor-B Via Recombinant Adeno-Associated Virus Resolves Chronic Ischemic Malperfusion: Role of Vessel Maturation

• Published in: Journal of the American College of Cardiology Vol. 56; no. 5; pp. 414 - 422
• Main Authors: KUPATT, Christian, HINKEL, Rabea, THEIN, Eckart, BANFI, Andrea, BOEKSTEGERS, Peter, PFOSSER, Achim, EL-AOUNI, Chiraz, WUCHRER, Alexander, FRITZ, Andrea, GLOBISCH, Franziska, THORMANN, Michael, HORSTKOTTE, Jan, LEBHERZ, Corinna
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier 27-07-2010; Elsevier Limited
• Subjects: Angiogenesis; Animals; Biological and medical sciences; Blood Vessels - pathology; Cardiology; Cardiology. Vascular system; Catheters; Clinical trials; Dependovirus - metabolism; Drug dosages; Experiments; Fixed rates; Gene Transfer Techniques; Genetic Techniques; Genetic Therapy - methods; Genetic Vectors; Heart; Ischemia; Ischemia - therapy; Laboratory animals; Medical sciences; Myocardium - pathology; Neovascularization, Physiologic; Plasmids; Polymerase chain reaction; Proteins; Proto-Oncogene Proteins c-sis - genetics; Proto-Oncogene Proteins c-sis - therapeutic use; Rabbits; Swine; Transfection; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor A - therapeutic use; Veins & arteries; Germany; Virus; Associated virus; Chronic; Vascular endothelium growth factor; Ischemia; Recombinant virus; Cardiovascular disease; Circulatory system; Cardiology; Platelet derived growth factor
• ISSN: 0735-1097; 1558-3597
• DOI: 10.1016/j.jacc.2010.03.050

We set out to investigate the ability of cardiotropic adeno-associated viral vector (AAV2.9 = recombinant adeno-associated virus [rAAV]) to induce prolonged expression of vascular endothelial growth factor (VEGF)-A and platelet-derived growth factor (PDGF)-B in a rabbit hindlimb ischemia model and a pig model of hibernating myocardium.
Gene therapy to induce angiogenesis and arteriogenesis has produced mixed results. However, long-acting viruses, such as rAAV, as well as combined induction of angiogenesis and vessel maturation might extend the therapeutic potential.
In rabbits, 0.5 x 10(11) particles rAAV.VEGF-A with or without 1 x 10(12) particles rAAV.PDGF-B were retroinfused at day 7 after femoral artery excision. At days 7 and 35, collateral counts and perfusion were determined, each value given as the day 35/day 7 ratio. Capillary-to-muscle fiber ratio was determined at day 35. In pigs, implantation of a reduction stent graft into the circumflex artery led to complete occlusion at day 28. At this time point, retroinfusion of rAAV.VEGF-A (1 x 10(13) particles), rAAV.VEGF-A/PDGF-B (2 x 10(12) and 4 x 10(12) particles, respectively) or mock transfection was performed. Ejection fraction and left ventricular end-diastolic pressure were assessed at days 28 and 56.
In rabbits, rAAV.VEGF-A strongly induced angiogenesis (capillary-to-muscle fiber ratio; 1.67 +/- 0.09 vs. 1.32 +/- 0.11 in rAAV.LacZ-treated limbs, p < 0.05), but not collateral growth (125 +/- 7% vs. 106 +/- 7%, p = NS) or perfusion (136 +/- 12% vs. 107 +/- 9%, p = NS). With VEGF-A/PDGF-B cotransfection, collateral growth increased to 146 +/- 9%, perfusion to 163 +/- 8% of the respective day 7 value (p < 0.05). In the pig model, retroinfusion of rAAV.VEGF-A/PDGF-B increased regional myocardial blood flow reserve from 101 +/- 4% (rAAV.Mock) to 129 +/- 8% (p < 0.05), based on collateral growth (3.2 +/- 0.3 in rAAV.Mock vs. 9.0 +/- 0.4 in rAAV.VEGF-A/PDGF-B, p < 0.05), whereas rAAV.VEGF-A did not alter flow reserve (112 +/- 7%) or collateral count (5.2 +/- 0.7). rAAV.VEGF-A/PDGF-B improved ejection fraction (55 +/- 5% vs. 34 +/- 3% in rAAV.Mock, p < 0.05) unlike rAAV.VEGF-A (37 +/- 2%).
Retroinfusion of rAAV.VEGF-A alone induces angiogenesis, but fails to enhance collateralization and perfusion, unless PDGF-B is cotransfected. In addition to neovascularization, rAAV.VEGF-A/PDGF-B improves regional and global myocardial function in hibernating myocardium.