Efficient and Durable Gene Transfer to Transplanted Heart Using Adeno-associated Virus 9 Vector

Miyagi, Naoto; Rao, Vinay P.; Ricci, Davide; Du, Zeji; Byrne, Guerard W.; Bailey, Kent R.; Nakai, Hiroyuki; Russell, Stephen J.; McGregor, Christopher G.A.

doi:10.1016/j.healun.2008.01.025

« Previous Next »The Journal of Heart and Lung Transplantation
Volume 27, Issue 5 , Pages 554-560, May 2008

Efficient and Durable Gene Transfer to Transplanted Heart Using Adeno-associated Virus 9 Vector

Naoto Miyagi, MD
- William J. von Liebig Transplant Center, Mayo Clinic, Rochester, Minnesota
Vinay P. Rao, MD
- William J. von Liebig Transplant Center, Mayo Clinic, Rochester, Minnesota
Davide Ricci, MD
- William J. von Liebig Transplant Center, Mayo Clinic, Rochester, Minnesota
Zeji Du, MD, PhD
- William J. von Liebig Transplant Center, Mayo Clinic, Rochester, Minnesota
Guerard W. Byrne, PhD
- William J. von Liebig Transplant Center, Mayo Clinic, Rochester, Minnesota
Kent R. Bailey, PhD
- Department of Biostatistics, Mayo Clinic, Rochester, Minnesota
Hiroyuki Nakai, MD, PhD
- Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania
Stephen J. Russell, MD, PhD
- Molecular Medicine Program, Mayo Clinic, Rochester, Minnesota.
Christopher G.A. McGregor, MB, FRCS, MD (Hons)
- William J. von Liebig Transplant Center, Mayo Clinic, Rochester, Minnesota
- Reprint requests: Christopher G. A. McGregor, MD (Hons), William J. von Liebig Transplant Center, Mayo Clinic College of Medicine, Mayo Clinic, 200 First Street SW, Rochester, MN 55905. Telephone: 507-255-6038. Fax: 507-255-4500.

Received 28 September 2007; received in revised form 14 January 2008; accepted 24 January 2008.

Background

In this investigation we studied the efficacy and durability of recombinant adeno-associated virus serotype 9 (rAAV9) vector–mediated gene transfer to the transplanted rat heart.

Methods

A rAAV9-CMV-lacZ vector diluted in cold (4°C) University of Wisconsin solution was used to perfuse the rat coronary vasculature for 20 minutes prior to syngeneic heterotopic transplantation. Perfusion experiments (six groups, n = 3/group) were performed without rAAV9 and at four separate doses ranging from 2 × 10⁹ to 2 × 10¹² viral genomes/ml. The transplanted heart was recovered 10 days or 3 months after transplantation and expression of lacZ assessed by histology, enzyme-linked immunoassay and real-time reverse transcript–polymerase chain reaction (RT-PCR). In a final group (n = 3), rAAV9 was administered systemically to compare the cardiac transduction efficiency and viral distribution to other organs.

Results

Transduction efficiency of perfused virus correlated with vector dose (p < 0.0001), with myocardial transduction ranging up to 71.74% at the highest dose. Cardiac expression of lacZ was equivalent at 10 days and 3 months. There was no evidence of viral gene transfer to other organs after heart transplantation.

Conclusions

Our findings demonstrate efficient and durable rAAV9-mediated gene transfer to the transplanted heart after ex vivo perfusion and suggest that AAV9 is a promising vector for cardiac gene therapy.