Targeting the respiratory muscles of fetal sheep for prenatal gene therapy for Duchenne muscular dystrophy

Weisz, Boaz; David, Anna L.; Gregory, Lisa G.; Perocheau, Dany; Ruthe, Ali; Waddington, Simon N.; Themis, Mike; Cook, Terry; Coutelle, Charles; Rodeck, Charles H.; Peebles, Donald M.

doi:10.1016/j.ajog.2005.06.077

« Previous Next »American Journal of Obstetrics & Gynecology
Volume 193, Issue 3, Supplement , Pages 1105-1109, September 2005

Targeting the respiratory muscles of fetal sheep for prenatal gene therapy for Duchenne muscular dystrophy

Boaz Weisz, MD
- Department of Obstetrics and Gynaecology, University College London
- Reprint requests: Dr Boaz Weisz, Department of Obstetrics and Gynaecology, University College London, 86-96 Chenies Mews, WC1E6HX, London, UK.
Anna L. David, MD
- Department of Obstetrics and Gynaecology, University College London
Lisa G. Gregory, PhD
- Imperial College of Science, Technology and Medicine, University of London, Section of Molecular Genetics, Gene Therapy Research Group
Dany Perocheau, BSc
- Department of Obstetrics and Gynaecology, University College London
Ali Ruthe, BSc
- Department of Obstetrics and Gynaecology, University College London
Simon N. Waddington, PhD
- Imperial College of Science, Technology and Medicine, University of London, Section of Molecular Genetics, Gene Therapy Research Group
Mike Themis, PhD
- Imperial College of Science, Technology and Medicine, University of London, Section of Molecular Genetics, Gene Therapy Research Group
Terry Cook, MD
- Department of Histopathology, Imperial College of Science, Technology and Medicine, University of London, London, United Kingdom
Charles Coutelle, PhD
- Imperial College of Science, Technology and Medicine, University of London, Section of Molecular Genetics, Gene Therapy Research Group
Charles H. Rodeck, MD
- Department of Obstetrics and Gynaecology, University College London
Donald M. Peebles, MD
- Department of Obstetrics and Gynaecology, University College London

Received 24 February 2005; received in revised form 29 April 2005; accepted 26 May 2005.

Objective

Duchenne muscular dystrophy (DMD) is a lethal degenerative muscular disease. Fetal gene therapy may correct the primary genetic defect. Our aim was to achieve expression of a reporter gene in the respiratory muscles of early gestation fetal sheep.

Study design

An adenovirus vector containing the β-galactosidase reporter gene (AdRSVβgal) was injected into the thoracic musculature (n=3) and pleural cavity (n=6) of fetal sheep (61-67 days' gestation) under ultrasound guidance. Tissues were harvested after 48 hours and site and intensity of β-galactosidase expression were assessed.

Results

Limited transgene expression observed after a single injection was improved by multiple injections, but remained localized. Ultrasound-guided creation of a hydrothorax led to an increase in the intensity of β-galactosidase expression (ELISA). X-gal staining and immunohistochemistry showed that vector spread was confined to the innermost intercostal musculature.