Split intein-mediated protein trans-splicing to express large dystrophins.

Autor: Tasfaout H; Department of Neurology, University of Washington School of Medicine, Seattle, WA, USA. tasfaout@uw.edu.; Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Washington School of Medicine, Seattle, WA, USA. tasfaout@uw.edu., Halbert CL; Department of Neurology, University of Washington School of Medicine, Seattle, WA, USA.; Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Washington School of Medicine, Seattle, WA, USA., McMillen TS; Department of Bioengineering, College of Engineering and School of Medicine, University of Washington, Seattle, WA, USA., Allen JM; Department of Neurology, University of Washington School of Medicine, Seattle, WA, USA.; Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Washington School of Medicine, Seattle, WA, USA., Reyes TR; Department of Neurology, University of Washington School of Medicine, Seattle, WA, USA.; Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Washington School of Medicine, Seattle, WA, USA., Flint GV; Department of Bioengineering, College of Engineering and School of Medicine, University of Washington, Seattle, WA, USA., Grimm D; Department of Infectious Diseases/Virology, Section Viral Vector Technologies, Medical Faculty and Faculty of Engineering Sciences, Center for Integrative Infectious Disease Research (CIID), University of Heidelberg, Heidelberg, Germany.; BioQuant, University of Heidelberg, Heidelberg, Germany.; German Center for Infection Research (DZIF) and German Center for Cardiovascular Research (DZHK), Heidelberg, Germany., Hauschka SD; Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Washington School of Medicine, Seattle, WA, USA.; Department of Biochemistry, University of Washington School of Medicine, Seattle, WA, USA., Regnier M; Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Washington School of Medicine, Seattle, WA, USA.; Department of Bioengineering, College of Engineering and School of Medicine, University of Washington, Seattle, WA, USA.; Center for Translational Muscle Research, University of Washington, Seattle, WA, USA., Chamberlain JS; Department of Neurology, University of Washington School of Medicine, Seattle, WA, USA. jsc5@uw.edu.; Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Washington School of Medicine, Seattle, WA, USA. jsc5@uw.edu.; Department of Biochemistry, University of Washington School of Medicine, Seattle, WA, USA. jsc5@uw.edu.; Center for Translational Muscle Research, University of Washington, Seattle, WA, USA. jsc5@uw.edu.
Jazyk: angličtina
Zdroj: Nature [Nature] 2024 Aug; Vol. 632 (8023), pp. 192-200. Date of Electronic Publication: 2024 Jul 17.
DOI: 10.1038/s41586-024-07710-8
Abstrakt: Gene replacement using adeno-associated virus (AAV) vectors is a promising therapeutic approach for many diseases 1,2 . However, this therapeutic modality is challenged by the packaging capacity of AAVs (approximately 4.7 kilobases) 3 , limiting its application for disorders involving large coding sequences, such as Duchenne muscular dystrophy, with a 14 kilobase messenger RNA. Here we developed a new method for expressing large dystrophins by utilizing the protein trans-splicing mechanism mediated by split inteins. We identified several split intein pairs that efficiently join two or three fragments to generate a large midi-dystrophin or the full-length protein. We show that delivery of two or three AAVs into dystrophic mice results in robust expression of large dystrophins and significant physiological improvements compared with micro-dystrophins. Moreover, using the potent myotropic AAVMYO 4 , we demonstrate that low total doses (2 × 10 13  viral genomes per kg) are sufficient to express large dystrophins in striated muscles body-wide with significant physiological corrections in dystrophic mice. Our data show a clear functional superiority of large dystrophins over micro-dystrophins that are being tested in clinical trials. This method could benefit many patients with Duchenne or Becker muscular dystrophy, regardless of genotype, and could be adapted to numerous other disorders caused by mutations in large genes that exceed the AAV capacity.
(© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)
Databáze: MEDLINE
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