RNA trans-splicing treatment for Duchenne muscular dystrophy
- 1Salk Institute for Biological Studies
Abstract
Duchenne muscular dystrophy (DMD) is a fatal disorder caused by dystrophin mutations, leading to progressive muscle degeneration and cardiac failure. Although AAV-based DMD therapies with innovative designs to overcome the cargo limits of the virus are effective in animal models, these current systems may face clinical translation challenges related to efficiency, off-target effects, and immunogenicity. We developed a multi-vector RNA End Joining (REJ) system to split large genes into multiple co-delivered AAVs that engage the cell's intrinsic spliceosome to precisely reassemble RNA segments that encode very large scar-free proteins. We show the system is efficient and has negligible off-target interactions. In vivo, REJ vectors expressing the adenine base editor Abe8e, mini-dystrophin Dp253, or native full-length dystrophin Dp427 each prevented muscle degeneration. Machine-learning histopathology of more than 120,000 myofibers revealed robust reductions in nuclear infiltration, improved centronucleation, and normalized hypertrophy, supported by functional, transcriptomic, and behavioral analyses. These findings establish REJ as a clinically viable AAV-based strategy for DMD that enables expression of large therapeutic proteins while avoiding potentially antigenic bacterial protein- or DNA-recombinases.
Related
- Planar covariation: a new key for the early detection of Duchenne muscular dystrophy walking behaviour
- Longitudinal measurements of MRI-T2 in boys with Duchenne muscular dystrophy: Effects of age and disease progression
- Feasibility and efficacy of personalized cardiorespiratory and strength exercise in Duchenne muscular dystrophy: a pilot study
- Generation and characterization of three human induced pluripotent stem cell lines from patients with Duchenne muscular dystrophy
- Physiological Reference Mapping Reveals Context-Dependent Dysregulation of Regenerative Programs in Duchenne Muscular Dystrophy.
BibTeX
@misc{hsu2026transsplicing,
title = {{RNA trans-splicing treatment for Duchenne muscular dystrophy}},
author = {R. H. Hsu and C. E. Williams and G. Maier and M. Gullo and K. Lettieri and C. J. Alvarez and K. J. Hermann and S. Kramer and S. Panda and L. C. Bachmann and S. L. Pfaff},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.13.751281},
url = {https://www.biorxiv.org/content/10.64898/2026.09.13.751281v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - RNA trans-splicing treatment for Duchenne muscular dystrophy
AU - R. H. Hsu
AU - C. E. Williams
AU - G. Maier
AU - M. Gullo
AU - K. Lettieri
AU - C. J. Alvarez
AU - K. J. Hermann
AU - S. Kramer
AU - S. Panda
AU - L. C. Bachmann
AU - S. L. Pfaff
PY - 2026
DA - 2026/09/13
PB - bioRxiv
DO - 10.64898/2026.09.13.751281
UR - https://www.biorxiv.org/content/10.64898/2026.09.13.751281v1
AB - Duchenne muscular dystrophy (DMD) is a fatal disorder caused by dystrophin mutations, leading to progressive muscle degeneration and cardiac failure. Although AAV-based DMD therapies with innovative designs to overcome the cargo limits of the virus are effective in animal models, these current systems may face clinical translation challenges related to efficiency, off-target effects, and immunogenicity. We developed a multi-vector RNA End Joining (REJ) system to split large genes into multiple co-delivered AAVs that engage the cell's intrinsic spliceosome to precisely reassemble RNA segments that encode very large scar-free proteins. We show the system is efficient and has negligible off-target interactions. In vivo, REJ vectors expressing the adenine base editor Abe8e, mini-dystrophin Dp253, or native full-length dystrophin Dp427 each prevented muscle degeneration. Machine-learning histopathology of more than 120,000 myofibers revealed robust reductions in nuclear infiltration, improved centronucleation, and normalized hypertrophy, supported by functional, transcriptomic, and behavioral analyses. These findings establish REJ as a clinically viable AAV-based strategy for DMD that enables expression of large therapeutic proteins while avoiding potentially antigenic bacterial protein- or DNA-recombinases.
ER -