1. ¹Program in Transplantation Biology, Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA
2. ²Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA
3. ³Department of Neurology, University of Washington, Seattle, Washington, USA
4. ⁴Department of Surgery, University of Washington, Seattle, Washington, USA
5. ⁵Department of Urology, University of Washington, Seattle, Washington, USA
6. ⁶Department of Biochemistry, University of Washington, Seattle, Washington, USA
7. ⁷Department of Medicine, University of Washington, Seattle, Washington, USA

Correspondence: Rainer Storb, Fred Hutchinson Cancer Research Center, Division of Clinical Research, 1100 Fairview Avenue N, D1-100, Seattle, Washington 98109, USA. E-mail: rstorb@fhcrc.org

Received 20 December 2006; Accepted 26 February 2007; Published online 10 April 2007.

Abstract

See page 1040

Adeno-associated virus–based vector (AAV)–mediated gene delivery has been successful in some animal models of human disease such as the mdx mouse model of human Duchenne muscular dystrophy (DMD). However, recent evidence of immune-mediated loss of vector persistence in dogs and humans suggests that immune modulation might be necessary to achieve successful long-term transgene expression in these species. We have previously demonstrated that direct intramuscular injection of AAV2 or AAV6 in wild-type random-bred dogs resulted in a robust immune response to capsid or capsid-associated proteins. We now demonstrate that a brief course of immunosuppression with a combination of anti-thymocyte globulin (ATG), cyclosporine (CSP), and mycophenolate mofetil (MMF) is sufficient to permit long-term and robust expression of a canine micro-dystrophin (c--dys) transgene in the skeletal muscle of a dog model for DMD (canine X-linked muscular dystrophy, or cxmd dog) and that its expression restored localization of components of the dystrophin-associated protein complex at the muscle membrane. This protocol has potential applications to human clinical trials to enhance AAV-mediated therapies.