Controlling Adenoviral Gene Transfer in Heart by Catheter-Based Coronary Perfusion

Abstract

This chapter discusses a catheter-based method for gene delivery that results in 58% efficient gene transfer to rat heart with minimal infection of peripheral organs. This infection approach is characterized by several features—the heart is entirely isolated in vivo by clamping all vessels to/from the heart simultaneously with a single clamp, a catheter positioned in the aortic root delivers the viral solution, while a second catheter positioned in the right ventricle provides a path for coronary efflux, the retrograde perfusion of the heart enables the blood to be flushed from the heart and replaced with well-oxygenated buffer containing the adenovirus and permeability agents; and at the end of the infection protocol, unsequestered virus is flushed from the heart prior to removing the clamp. Continuously perfusing the myocardium can control the viral concentration, permeability agent, cardioplegic solution, and temperature of the heart versus the body. Myocardial gene therapy represents a promising approach for the treatment of inherited heart diseases, cardiomyopathies, and congestive heart failure. Extensive work has demonstrated that recombinant adenoviral vectors can efficiently transduce cardiomyocytes in vivo. However, targeting gene expression to the heart and controlling the level of transfer has been a challenge. In this study, we describe a catheter-based method for gene delivery that results in 58% efficient gene transfer to rat heart with minimal infection of peripheral organs. The heart is completely isolated in vivo during delivery, and unsequestered virus is flushed from the heart prior to releasing the cross-clamp. We compare the results to existing methods and highlight promising new strategies.