Abstract
Duchenne muscular dystrophy (DMD) is a muscle wasting disorder caused by mutations in the DMD gene. Restoration of full-length dystrophin protein in skeletal muscle would have therapeutic benefit, but lentivirally mediated delivery of such a large gene in vivo has been hindered by lack of tissue specificity, limited transduction, and insufficient transgene expression. To address these problems, we developed a lentiviral vector, which contains a muscle-specific promoter and sequence-optimized full-length dystrophin, to constrain dystrophin expression to differentiated myotubes/myofibers and enhance the transgene expression. We further explored the efficiency of restoration of full-length dystrophin in vivo, by grafting DMD myoblasts that had been corrected by this optimized lentiviral vector intramuscularly into an immunodeficient DMD mouse model. We show that these lentivirally corrected DMD myoblasts effectively reconstituted full-length dystrophin expression in 93.58% ± 2.17% of the myotubes in vitro. Moreover, dystrophin was restored in 64.4% ± 2.87% of the donor-derived regenerated muscle fibers in vivo, which were able to recruit members of the dystrophin-glycoprotein complex at the sarcolemma. This study represents a significant advance over existing cell-mediated gene therapy strategies for DMD that aim to restore full-length dystrophin expression in skeletal muscle.
| Original language | English |
|---|---|
| Pages (from-to) | 491-507 |
| Number of pages | 17 |
| Journal | Molecular Therapy - Methods and Clinical Development |
| Volume | 25 |
| DOIs | |
| Publication status | Published - 17 May 2022 |
| Externally published | Yes |
Bibliographical note
Funding Information:This work was funded by MDUK (grant 17GRO-PG36-0165 ) and in part by the Wellcome Trust (grant 210774/Z/18/ ). For the purpose of open access, the author has applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. The support of the MRC Center for Neuromuscular Diseases Biobank is gratefully acknowledged. J.E.M. was supported by the Great Ormond Street Hospital Children Charity . This research was supported by the NIHR Great Ormond Street Hospital Biomedical Research Centre . The views expressed are those of the authors and not necessarily those of the NHS , the NIHR , or the Department of Health .
Funding Information:
This work was funded by MDUK (grant 17GRO-PG36-0165) and in part by the Wellcome Trust (grant 210774/Z/18/). For the purpose of open access, the author has applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. The support of the MRC Center for Neuromuscular Diseases Biobank is gratefully acknowledged. J.E.M. was supported by the Great Ormond Street Hospital Children Charity. This research was supported by the NIHR Great Ormond Street Hospital Biomedical Research Centre. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health. Conceptualization and designing of the project, J.M. L.P. and J.E.M.; methodology, J.M. and J.C.; analysis and investigation, J.M. and M.M.; writing – original draft preparation, J.M. and J.E.M.; writing – review and editing, J.E.M. M.M. J.C. L.P. and F.M.; supervision, J.E.M.; project administration, J.E.M.; funding acquisition, J.E.M. Royal Holloway University of London has a patent on the sequence-optimized full-length DMD cDNA.
Publisher Copyright:
© 2022 The Author(s)