We use a novel technique that allows for closed recirculation of vector genomes in the cardiac circulation using cardiopulmonary bypass, referred to here as molecular cardiac surgery with recirculating delivery (MCARD). We demonstrate that this platform technology is highly efficient in isolating the heart from the systemic circulation in vivo. Using MCARD, we compare the relative efficacy of single-stranded (ss) adeno-associated virus (AAV)6, ssAAV9 and self-complimentary (sc)AAV6-encoding enhanced green fluorescent protein, driven by the constitutive cytomegalovirus promoter to transduce the ovine myocardium in situ. MCARD allows for the unprecedented delivery of up to 48 green fluorescent protein genome copies per cell globally in the sheep left ventricular (LV) myocardium. We demonstrate that scAAV6-mediated MCARD delivery results in global, cardiac-specific LV gene expression in the ovine heart and provides for considerably more robust and cardiac-specific gene delivery than other available delivery techniques such as intramuscular injection or intracoronary injection; thus, representing a potential, clinically translatable platform for heart failure gene therapy.
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This work was sponsored in part by the National Heart Lung and Blood Institute (1-R01-HL083078-01A2), CR Bridges, PI; we would like to acknowledge the Gene Therapy Resource Program (GTRP) of the National Heart, Lung and Blood Institute, National Institutes of Health for providing a subset of the gene vectors used in this study and the Immunology Core of the Gene Therapy Program at the University of Pennsylvania, supported by NIH P30-DK047757 (PI: James Wilson). The authors have no proprietary disclosures to report. We thank the animal care staff and technicians for excellent care and handling of the animals in the Biomedical Research Building Vivarium-University of Pennsylvania.
The authors declare no conflict of interest.
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White, J., Thesier, D., Swain, J. et al. Myocardial gene delivery using molecular cardiac surgery with recombinant adeno-associated virus vectors in vivo. Gene Ther 18, 546–552 (2011). https://doi.org/10.1038/gt.2010.168
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