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Cardio-specific long-term gene expression in a porcine model after selective pressure-regulated retroinfusion of adeno-associated viral (AAV) vectors

Abstract

Cornerstone for an efficient cardiac gene therapy is the need for a vector system, which enables selective and long-term expression of the gene of interest. In rodent animal models adeno-associated viral (AAV) vectors like AAV-6 have been shown to efficiently transduce cardiomyocytes. However, since significant species-dependent differences in transduction characteristics exist, large animal models are of imminent need for preclinical evaluations. We compared gene transfer efficiencies of AAV-6 and heparin binding site-deleted AAV-2 vectors in a porcine model. Application of the AAVs was performed by pressure-regulated retroinfusion of the anterior interventricular cardiac vein, which has been previously shown to efficiently deliver genes to the myocardium (3.5 × 1010 viral genomes per animal; n=5 animals per group). All vectors harbored a luciferase reporter gene under control of a cytomegalovirus (CMV)-enhanced 1.5 kb rat myosin light chain promoter (CMV-MLC2v). Expression levels were evaluated 4 weeks after gene transfer by determining luciferase activities. To rule out a systemic spillover peripheral tissue was analyzed by PCR for the presence of vector genomes. Selective retroinfusion of AAV serotype 6 vectors into the anterior cardiac vein substantially increased reporter gene expression in the targeted distal left anterior descending (LAD) territory (65 943±31 122 vs control territory 294±69, P<0.05). Retroinfusion of AAV-2 vectors showed lower transgene expression, which could be increased with coadministration of recombinant human vascular endothelial growth factor (1365±707 no vascular endothelial growth factor (VEGF) vs 38 760±2448 with VEGF, P<0.05). Significant transgene expression was not detected in other organs than the heart, although vector genomes were detected also in the lung and liver. Thus, selective retroinfusion of AAV-6 into the coronary vein led to efficient long-term myocardial reporter gene expression in the targeted LAD area of the porcine heart. Coapplication of VEGF significantly increased transduction efficiency of AAV-2.

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Acknowledgements

We gratefully acknowledge the excellent technical support of Elisabeth Ronft and Susanne Helbig from the Department of Internal Medicine I, Grosshadern University Hospital. We thank Barbara Leuchs, Renate Eudenbach and the Vector Core Facility of the German Cancer Research Center for production of high-titer vector stocks. This work was supported in part by Program for Funding of Science and Teaching of the University of Munich research grant 412 (to PWR) and a grant of the Bundesministerium für Bildung und Forschung (01GU0525 to PWR and PB and 01GU0527 to OJM, JAK and HAK).

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Correspondence to P Boekstegers.

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Raake, P., Hinkel, R., Müller, S. et al. Cardio-specific long-term gene expression in a porcine model after selective pressure-regulated retroinfusion of adeno-associated viral (AAV) vectors. Gene Ther 15, 12–17 (2008). https://doi.org/10.1038/sj.gt.3303035

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  • DOI: https://doi.org/10.1038/sj.gt.3303035

Keywords

  • heart
  • AAV
  • retroinfusion
  • VEGF

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