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

Abstract

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 × 1014 (high), 5 × 1013 (medium), or 5 × 1012 (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.

• Authors: Chady H Hakim, Nathalie Clément, Lakmini P Wasala, Hsiao T Yang, Yongping Yue, Keqing Zhang, Kasun Kodippili, Laura Adamson-Small, Xiufang Pan, Joel S Schneider, N Nora Yang, Jeffrey S Chamberlain, Barry J Byrne, Dongsheng Duan
• Journal: Molecular therapy. Methods & clinical development (Mol Ther Methods Clin Dev) Vol. 18 Pg. 664-678 (Sep 11 2020) ISSN: 2329-0501 [Print] United States
• PMID: 32775499 (Publication Type: Journal Article)
• Copyright: © 2020 The Author(s).