Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Myocardial gene delivery using molecular cardiac surgery with recombinant adeno-associated virus vectors in vivo


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.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7


  1. Lloyd-Jones DM, Larson MG, Leip EP, Beiser A, D’Agostino RB, Kannel WB et al. Lifetime risk for developing congestive heart failure: the Framingham Heart Study. Circulation 2002; 106: 3068–3072.

    Article  PubMed  Google Scholar 

  2. Lloyd-Jones DM, Adams R, Carnethon M, De Simone J, Ferguson TB, Flegal K et al. Heart Disease and Stroke Statistics- 2009 update. A report from the American Heart Association statistics committee and stroke statistics subcommittee. Circulation 2009; 119: 480–486.

    Article  PubMed  Google Scholar 

  3. Gregorevic P, Blankinship MJ, Allen JM, Crawford RW, Meuse L, Miller DG et al. Systemic delivery of genes to striated muscles using adeno-associated viral vectors. Nat Med 2004; 10: 828–834.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wang Z, Zhu T, Qiao C, Zhou L, Wang B, Zhang J et al. Adeno-associated virus serotype 8 efficiently delivers genes to muscle and heart. Nat Biotechnol 2005; 23: 321–328.

    Article  CAS  PubMed  Google Scholar 

  5. Inagaki K, Fuess S, Storm TA, Gibson GA, Mctiernan CF, Kay MA et al. Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8. Mol Ther 2006; 14: 45–53.

    Article  CAS  PubMed  Google Scholar 

  6. Bridges CR . ‘Recirculating cardiac delivery’ method of gene delivery should be called ‘non-recirculating’ method [letter]. Gene Therapy 2009; 16: 939–940.

    Article  CAS  PubMed  Google Scholar 

  7. Byrne MJ, Power JM, Preovolos A, Mariani JA, Hajjar RJ, Kaye DM . Recirculating cardiac delivery of AAV2/1SERCA2a improves myocardial function on an experimental model of heart failure in large animals. Gene Therapy 2008; 15: 1550–1557.

    Article  CAS  PubMed  Google Scholar 

  8. French BA, Mazur W, Geske RS, Bolli R . Direct in vivo gene transfer into porcine myocardium using replication-deficient adenoviral vectors. Circulation 1994; 90: 2414–2424.

    Article  CAS  PubMed  Google Scholar 

  9. Bridges CR, Gopal K, Holt DE, Yarnall C, Cole S, Anderson RB et al. Efficient myocyte gene delivery with complete cardiac surgical isolation in situ. J Thorac Cardiovasc Surg 2005; 130: 1364–1370.

    Article  PubMed  Google Scholar 

  10. Rodgers RJ, O’Shea JD, Bruce NW . Morphometric analysis of the ovine corpus luteum. J Anat 1984; 138: 757–770.

    PubMed Central  PubMed  Google Scholar 

  11. Wang L, Figueredo J, Calcedo R, Wilson JM . Cross-presentation of adeno-associated virus serotype 2 capsids activates cytotoxic T cells but does not render hepatocytes effective cytolytic targets. Hum Gene Ther 2007; 18: 185–194.

    Article  CAS  PubMed  Google Scholar 

  12. Gao GP, Lu Y, Sun X, Johnston J, Calcedo R, Grant R et al. High-level transgene expression in nonhuman primate liver with novel adeno-associated virus serotypes containing self-complementary genomes. J Virol 2006; 80: 6192–6194.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Hajjar RJ, Samulski RJ . A Silver bullet to treat heart failure. Gene Therapy 2006; 13: 997.

    Article  CAS  PubMed  Google Scholar 

  14. Hajjar RJ, Zsebo K, Deckelbaum L, Thompson C, Rudy J, Yaroshinsky A et al. Design of a phase 1/2 trial of intracoronary administration of AAV1/SERCA2a in patients with heart failure. J Card Fail 2008; 14: 355–367.

    Article  CAS  PubMed  Google Scholar 

  15. Rosengart TK, Lee LY, Patel SR, Sanborn TA, Parikh M, Bergman GW et al. Angiogenesis gene therapy: phase 1 assessment of direct intramyocardial administration of an adenovirus vector expressing VEGF121 cDNA to individuals with clinically significant severe coronary artery disease. Circulation 1999; 100: 468–474.

    Article  CAS  PubMed  Google Scholar 

  16. Grossman PM, Han Z, Palasis M, Barry JJ, Lederman RJ . Incomplete retention after direct myocardial injection. Cathet Cardiovasc Intervent 2002; 55: 392–397.

    Article  Google Scholar 

  17. Von Harsdorf R, Schott RJ, Shen YT, Vatner SF, Mahdavi V, Nadal-Ginard B . Gene injection into canine myocardium as a useful model for studying expression in the heart of large mammals. Circ Res 1993; 72: 688–695.

    Article  CAS  PubMed  Google Scholar 

  18. Asokan A, Conway JC, Phillips JL, Li C, Hegge J, Sinnott R et al. Reengineering a receptor footprint of adeno-associated virus enables selective and systemic gene transfer to muscle. Nat Biotechnol 2010; 28: 79–82.

    Article  CAS  PubMed  Google Scholar 

  19. Zolotukhin S, Potter M, Hauswirth W, Guy J, Muzyczka N . T ‘humanized’ green fluorescent protein cDNA adapted for high-level expression in mammalian cells. J Virol 1996; 70: 4646–4654.

    CAS  PubMed Central  PubMed  Google Scholar 

  20. McCarty D, Monahan P, Samulski RJ . Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis. Gene Therapy 2001; 8: 1248–1254.

    Article  CAS  PubMed  Google Scholar 

  21. Grieger J, Choi V, Samulski RJ . Production and characterization of adeno-associated viral vectors. Nat Protoc 2006; 1: 1412–1428.

    Article  CAS  PubMed  Google Scholar 

  22. Zolotuchin S, Byrne BJ, Mason E, Zolotuchin I, Potter M, Chestnut K et al. Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Therapy 1999; 6: 973–985.

    Article  Google Scholar 

Download references


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.

Author information

Authors and Affiliations


Corresponding author

Correspondence to C R Bridges.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • AAV vectors
  • molecular cardiac surgery
  • cardiac gene therapy

This article is cited by


Quick links