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Repeated, but not single, VEGF gene transfer affords protection against ischemic muscle lesions in rabbits with hindlimb ischemia

Gene Therapy volume 16, pages 716–723 (2009)Cite this article

Abstract

Vascular endothelial growth factor (VEGF) gene transfer-mediated angiogenesis has been proposed for peripheral artery disease. However, protocols using single administration have shown little benefit. Given that the transient nature of VEGF gene expression provokes instability of neovasculature, we hypothesized that repeated administration would provide efficient tissue protection. We thus compared single vs repeated transfection in a rabbit model of hindlimb ischemia by injecting a plasmid encoding human VEGF165 (pVEGF165) at 7 (GI, n=10) or 7 and 21 (GII, n=10) days after surgery. Placebo animals (GIII, n=10) received empty plasmid. Fifty days after surgery, single and repeated administration similarly increased saphenous peak flow velocity and quantity of angiographically visible collaterals. However, microvasculature increased only with repeated transfection: capillary density was 49.4±15.4 capillaries per 100 myocytes in GI, 84.6±14.7 in GII (P<0.01 vs GI and GIII) and 49.3±13.6 in GIII, and arteriolar density was 1.9±0.6 arterioles per mm2 in GI, 3.0±0.9 in GII (P<0.01 vs GI and GIII) and 1.5±0.6 in GIII. Muscle lesions were reduced only within repeated transfection. With single administration, gene expression peaked at 7 days and declined rapidly, but with repeated administration, it remained positive at 50 days. At 90 days of repeated transfection (additional animals), gene expression decreased significantly, but neovessel densities did not. Thus, repeated, but not single, VEGF gene transfection resulted in increased microvasculature, which, in turn, afforded effective protection against ischemic muscle damage.

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Acknowledgements

We are grateful to medical student Anna Hnatiuk and technicians, Julio Martínez, Fabián Gauna and Luis Palacios, for surgical and morphometric analysis assistance. We also thank veterinarians, María Inés Besansón, Pedro Iguain and Marta Tealdo, for anesthetic management, and animal house assistants, Juan Ocampo, Osvaldo Sosa and Juan Carlos Mansilla, for animal care. This work was supported by Grant BID 1728/OC-AR PID 268 from the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) of Argentina. AC, RL and LC are established investigators of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina. AB, MC and CM are scientists of Bio Sidus, the biotechnological company developing the plasmid used in this report.

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Authors and Affiliations

  1. Department of Physiology, Favaloro University, Buenos Aires, Argentina

    F D Olea, G Vera Janavel & A Crottogini

  2. Department of Pathology, Favaloro University, Buenos Aires, Argentina

    L Cuniberti, G Yannarelli, P Cabeza Meckert & R Laguens

  3. Scientific Investigation Commission, Buenos Aires Province, Argentina

    P Cabeza Meckert

  4. Favaloro Foundation University Hospital, Buenos Aires, Argentina

    J Cors, L Valdivieso, G Lev & O Mendiz

  5. Bio Sidus, Buenos Aires, Argentina

    A Bercovich, M Criscuolo & C Melo

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  1. F D Olea
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Correspondence to A Crottogini.

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Olea, F., Janavel, G., Cuniberti, L. et al. Repeated, but not single, VEGF gene transfer affords protection against ischemic muscle lesions in rabbits with hindlimb ischemia. Gene Ther 16, 716–723 (2009). https://doi.org/10.1038/gt.2009.30

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