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

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

doi:10.1016/j.ajog.2005.06.077

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

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

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

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. Conclusion: Ultrasound-guided injection can deliver gene therapy vectors to the fetal pleural cavity and achieve transduction of the respiratory muscles.

Original language	English
Pages (from-to)	1105-1109
Number of pages	5
Journal	American Journal of Obstetrics and Gynecology
Volume	193
Issue number	3 SUPPL.
DOIs	https://doi.org/10.1016/j.ajog.2005.06.077
State	Published - Sep 2005
Externally published	Yes

Funding

Funders	Funder number
British Medical Research Council
Katharine Dormandy Haemophilia Trust

Keywords

Duchenne muscular dystrophy
Fetal gene therapy
Fetal medicine
Gene therapy
Ultrasound

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10.1016/j.ajog.2005.06.077

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Weisz, B., David, A. L., Gregory, L. G., Perocheau, D., Ruthe, A., Waddington, S. N., Themis, M., Cook, T., Coutelle, C., Rodeck, C. H., & Peebles, D. M. (2005). Targeting the respiratory muscles of fetal sheep for prenatal gene therapy for Duchenne muscular dystrophy. American Journal of Obstetrics and Gynecology, 193(3 SUPPL.), 1105-1109. https://doi.org/10.1016/j.ajog.2005.06.077

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title = "Targeting the respiratory muscles of fetal sheep for prenatal gene therapy for Duchenne muscular dystrophy",

abstract = "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. Conclusion: Ultrasound-guided injection can deliver gene therapy vectors to the fetal pleural cavity and achieve transduction of the respiratory muscles.",

keywords = "Duchenne muscular dystrophy, Fetal gene therapy, Fetal medicine, Gene therapy, Ultrasound",

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

note = "Funding Information: This work is supported by the British Medical Research Council. S. W. is supported by the Katharine Dormandy Haemophilia Trust. We wish to express our gratitude to Dr Jill Maddox, Centre for Animal Biotechnology, University of Melbourne Australia, for supplying primers to endogenous sheep sequences. ",

year = "2005",

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doi = "10.1016/j.ajog.2005.06.077",

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journal = "American Journal of Obstetrics and Gynecology",

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Weisz, B, David, AL, Gregory, LG, Perocheau, D, Ruthe, A, Waddington, SN, Themis, M, Cook, T, Coutelle, C, Rodeck, CH & Peebles, DM 2005, 'Targeting the respiratory muscles of fetal sheep for prenatal gene therapy for Duchenne muscular dystrophy', American Journal of Obstetrics and Gynecology, vol. 193, no. 3 SUPPL., pp. 1105-1109. https://doi.org/10.1016/j.ajog.2005.06.077

TY - JOUR

T1 - Targeting the respiratory muscles of fetal sheep for prenatal gene therapy for Duchenne muscular dystrophy

AU - Weisz, Boaz

AU - David, Anna L.

AU - Gregory, Lisa G.

AU - Perocheau, Dany

AU - Ruthe, Ali

AU - Waddington, Simon N.

AU - Themis, Mike

AU - Cook, Terry

AU - Coutelle, Charles

AU - Rodeck, Charles H.

AU - Peebles, Donald M.

N1 - Funding Information: This work is supported by the British Medical Research Council. S. W. is supported by the Katharine Dormandy Haemophilia Trust. We wish to express our gratitude to Dr Jill Maddox, Centre for Animal Biotechnology, University of Melbourne Australia, for supplying primers to endogenous sheep sequences.

PY - 2005/9

Y1 - 2005/9

N2 - 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. Conclusion: Ultrasound-guided injection can deliver gene therapy vectors to the fetal pleural cavity and achieve transduction of the respiratory muscles.

AB - 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. Conclusion: Ultrasound-guided injection can deliver gene therapy vectors to the fetal pleural cavity and achieve transduction of the respiratory muscles.

KW - Duchenne muscular dystrophy

KW - Fetal gene therapy

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KW - Gene therapy

KW - Ultrasound

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IS - 3 SUPPL.

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

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