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Efficient, cardiac-specific adenoviral gene transfer in rat heart by isolated retrograde perfusion in vivo

Gene Therapy volume 12, pages 958–964 (2005)Cite this article

Abstract

While a number of virus-based delivery schemes have been developed for myocardial gene transfer, no technique has proven capable of modifying a majority of cardiac myocytes rapidly and homogeneously in the in vivo rat model, and most schemes result in significant infection of the liver and other organs. However, adenoviral delivery to the excised heart during retrograde perfusion can produce 67–92% efficient gene transfer. In this study, we adapt this isolation/perfusion scheme to the heart in vivo. We isolated the heart in vivo by simultaneously clamping all vessels to/from the heart. The heart was then continuously retrograde perfused through a catheter positioned in the aortic root. A second catheter in the right ventricle provided a path for efflux. After perfusing the heart for 7.5 min with calcium-free Tyrode solution followed by 90 s no-flow viral exposure (AdV.cmv.LacZ; 1012 viral particles/ml), gene transfer efficiency was 60% compared to 5% by a conventional cross-clamp technique. Infection of peripheral organs was dramatically reduced. Given the prevalence of the rat in so many models of heart disease, this enhancement of infection represents an advancement in viral-based delivery of exogenous genes to heart for the study of gene therapy in vivo.

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Acknowledgements

We thank Drs David Geenen and Steven Pogwizd (UIC, Cardiology) for helpful discussion. This study was supported by grants from the American Heart Association (O'Donnell; 0230099N) and the National Heart, Lung, and Blood Institute, NIH (Lewandowski; RO1HL 62702, RO1HL 56178, R37HL49244).

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  1. Program in Integrative Cardiac Metabolism, Department of Physiology and Biophysics, University of Illinois at Chicago, College of Medicine, Chicago, IL, USA

    J Michael O'Donnell & E Douglas Lewandowski

  2. Medicine (Cardiology), University of Illinois at Chicago, College of Medicine, Chicago, IL, USA

    E Douglas Lewandowski

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  1. J Michael O'Donnell
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O'Donnell, J., Lewandowski, E. Efficient, cardiac-specific adenoviral gene transfer in rat heart by isolated retrograde perfusion in vivo. Gene Ther 12, 958–964 (2005). https://doi.org/10.1038/sj.gt.3302477

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