Long-term Restoration of Cardiac Dystrophin Expression in Golden Retriever Muscular Dystrophy Following RAAV6-mediated Exon Skipping

Overview

Journal Mol Ther

Publisher Cell Press

Specialties Molecular Biology
Pharmacology

Date 2011 Dec 8

PMID 22146342

Citations 49

Authors

Lawrence T Bish

Meg M Sleeper

Sean C Forbes

Bingjing Wang

Caryn Reynolds

Gretchen E Singletary

Dennis Trafny

Kevin J Morine

Julio Sanmiguel

Sylvain Cecchini

Tamas Virag

Adeline Vulin

Cyriaque Beley

Janet Bogan

James M Wilson

Krista Vandenborne

Joe N Kornegay

Glenn A Walter

Robert M Kotin

Luis Garcia

H Lee Sweeney

Affiliations

Soon will be listed here.

Abstract

Although restoration of dystrophin expression via exon skipping in both cardiac and skeletal muscle has been successfully demonstrated in the mdx mouse, restoration of cardiac dystrophin expression in large animal models of Duchenne muscular dystrophy (DMD) has proven to be a challenge. In large animals, investigators have focused on using intravenous injection of antisense oligonucleotides (AO) to mediate exon skipping. In this study, we sought to optimize restoration of cardiac dystrophin expression in the golden retriever muscular dystrophy (GRMD) model using percutaneous transendocardial delivery of recombinant AAV6 (rAAV6) to deliver a modified U7 small nuclear RNA (snRNA) carrying antisense sequence to target the exon splicing enhancers of exons 6 and 8 and correct the disrupted reading frame. We demonstrate restoration of cardiac dystrophin expression at 13 months confirmed by reverse transcription-PCR (RT-PCR) and immunoblot as well as membrane localization by immunohistochemistry. This was accompanied by improved cardiac function as assessed by cardiac magnetic resonance imaging (MRI). Percutaneous transendocardial delivery of rAAV6 expressing a modified U7 exon skipping construct is a safe, effective method for restoration of dystrophin expression and improvement of cardiac function in the GRMD canine and may be easily translatable to human DMD patients.

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