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Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease

Vector production scale-up is a major barrier in systemic adeno-associated virus (AAV) gene therapy. Many scalable manufacturing methods have been developed. However, the potency of the vectors generated by these methods has rarely been compared with vectors made by transient transfection (TT), the...

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Autores principales: Hakim, Chady H., Clément, Nathalie, Wasala, Lakmini P., Yang, Hsiao T., Yue, Yongping, Zhang, Keqing, Kodippili, Kasun, Adamson-Small, Laura, Pan, Xiufang, Schneider, Joel S., Yang, N. Nora, Chamberlain, Jeffrey S., Byrne, Barry J., Duan, Dongsheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7403893/
https://www.ncbi.nlm.nih.gov/pubmed/32775499
http://dx.doi.org/10.1016/j.omtm.2020.07.004
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author Hakim, Chady H.
Clément, Nathalie
Wasala, Lakmini P.
Yang, Hsiao T.
Yue, Yongping
Zhang, Keqing
Kodippili, Kasun
Adamson-Small, Laura
Pan, Xiufang
Schneider, Joel S.
Yang, N. Nora
Chamberlain, Jeffrey S.
Byrne, Barry J.
Duan, Dongsheng
author_facet Hakim, Chady H.
Clément, Nathalie
Wasala, Lakmini P.
Yang, Hsiao T.
Yue, Yongping
Zhang, Keqing
Kodippili, Kasun
Adamson-Small, Laura
Pan, Xiufang
Schneider, Joel S.
Yang, N. Nora
Chamberlain, Jeffrey S.
Byrne, Barry J.
Duan, Dongsheng
author_sort Hakim, Chady H.
collection PubMed
description Vector production scale-up is a major barrier in systemic adeno-associated virus (AAV) gene therapy. Many scalable manufacturing methods have been developed. However, the potency of the vectors generated by these methods has rarely been compared with vectors made by transient transfection (TT), the most commonly used method in preclinical studies. In this study, we blindly compared therapeutic efficacy of an AAV9 micro-dystrophin vector generated by the TT method and scalable herpes simplex virus (HSV) system in a Duchenne muscular dystrophy mouse model. AAV was injected intravenously at 5 × 10(14) (high), 5 × 10(13) (medium), or 5 × 10(12) (low) viral genomes (vg)/kg. Comparable levels of micro-dystrophin expression were observed at each dose in a dose-dependent manner irrespective of the manufacturing method. Vector biodistribution was similar in mice injected with either the TT or the HSV method AAV. Evaluation of muscle degeneration/regeneration showed equivalent protection by vectors made by either method in a dose-dependent manner. Muscle function was similarly improved in a dose-dependent manner irrespective of the vector production method. No apparent toxicity was observed in any mouse. Collectively, our results suggest that the biological potency of the AAV micro-dystrophin vector made by the scalable HSV method is comparable to that made by the TT method.
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spelling pubmed-74038932020-08-07 Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease Hakim, Chady H. Clément, Nathalie Wasala, Lakmini P. Yang, Hsiao T. Yue, Yongping Zhang, Keqing Kodippili, Kasun Adamson-Small, Laura Pan, Xiufang Schneider, Joel S. Yang, N. Nora Chamberlain, Jeffrey S. Byrne, Barry J. Duan, Dongsheng Mol Ther Methods Clin Dev Article Vector production scale-up is a major barrier in systemic adeno-associated virus (AAV) gene therapy. Many scalable manufacturing methods have been developed. However, the potency of the vectors generated by these methods has rarely been compared with vectors made by transient transfection (TT), the most commonly used method in preclinical studies. In this study, we blindly compared therapeutic efficacy of an AAV9 micro-dystrophin vector generated by the TT method and scalable herpes simplex virus (HSV) system in a Duchenne muscular dystrophy mouse model. AAV was injected intravenously at 5 × 10(14) (high), 5 × 10(13) (medium), or 5 × 10(12) (low) viral genomes (vg)/kg. Comparable levels of micro-dystrophin expression were observed at each dose in a dose-dependent manner irrespective of the manufacturing method. Vector biodistribution was similar in mice injected with either the TT or the HSV method AAV. Evaluation of muscle degeneration/regeneration showed equivalent protection by vectors made by either method in a dose-dependent manner. Muscle function was similarly improved in a dose-dependent manner irrespective of the vector production method. No apparent toxicity was observed in any mouse. Collectively, our results suggest that the biological potency of the AAV micro-dystrophin vector made by the scalable HSV method is comparable to that made by the TT method. American Society of Gene & Cell Therapy 2020-07-09 /pmc/articles/PMC7403893/ /pubmed/32775499 http://dx.doi.org/10.1016/j.omtm.2020.07.004 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Hakim, Chady H.
Clément, Nathalie
Wasala, Lakmini P.
Yang, Hsiao T.
Yue, Yongping
Zhang, Keqing
Kodippili, Kasun
Adamson-Small, Laura
Pan, Xiufang
Schneider, Joel S.
Yang, N. Nora
Chamberlain, Jeffrey S.
Byrne, Barry J.
Duan, Dongsheng
Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease
title Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease
title_full Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease
title_fullStr Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease
title_full_unstemmed Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease
title_short Micro-dystrophin AAV Vectors Made by Transient Transfection and Herpesvirus System Are Equally Potent in Treating mdx Mouse Muscle Disease
title_sort micro-dystrophin aav vectors made by transient transfection and herpesvirus system are equally potent in treating mdx mouse muscle disease
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7403893/
https://www.ncbi.nlm.nih.gov/pubmed/32775499
http://dx.doi.org/10.1016/j.omtm.2020.07.004
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