An attractive, therapeutic strategy for Duchenne muscular dystrophy (DMD) is to induce exon skipping in affected muscle cells, in order to restore the open reading frame of the dystrophin gene. Such an approach has been made in preclinical and clinical studies, using antisense oligonucleotides (AONs), however many hurdles remain due to the narrow therapeutic range of AONs. AON delivery is hampered by its short period of transient correction in dystrophin expression, and the chronically high dose it requires to achieve a sustained benefit in DMD patients. We used engineered stem cells as vectors for a permanent and efficient delivery of AONs. We engineered human DMD stem cells with a lentivirus, which permanently expresses the cloned AONs, and transplanted them into scid/mdx mice, the animal model of DMD. These engineered stem cells were fused with myoblasts and skeletal muscles of dystrophic scid/mdx mice, rescuing their dystrophin expression. We assessed that functional AONs were released in exosomes by these engineered, human DMD stem cells. These results demonstrated a bystander effect, mediated by the use of engineered stem cells for the delivery of AONs, and provided a possible new treatment method for DMD
Stem cell-mediated exon skipping of the dystrophin gene by the bystander effect / Y. Torrente, M. Meregalli, C. Beley, A. Farini, C. Sitzia, P. Razini, L. Cassinelli, F. Colleoni, A. Tavelli, M. Belicchi, L. Garcia. ((Intervento presentato al convegno Global Controls in Stem Cells tenutosi a Singapore nel 2014.
Stem cell-mediated exon skipping of the dystrophin gene by the bystander effect
Y. Torrente;M. Meregalli;A. Farini;M. Belicchi;
2014
Abstract
An attractive, therapeutic strategy for Duchenne muscular dystrophy (DMD) is to induce exon skipping in affected muscle cells, in order to restore the open reading frame of the dystrophin gene. Such an approach has been made in preclinical and clinical studies, using antisense oligonucleotides (AONs), however many hurdles remain due to the narrow therapeutic range of AONs. AON delivery is hampered by its short period of transient correction in dystrophin expression, and the chronically high dose it requires to achieve a sustained benefit in DMD patients. We used engineered stem cells as vectors for a permanent and efficient delivery of AONs. We engineered human DMD stem cells with a lentivirus, which permanently expresses the cloned AONs, and transplanted them into scid/mdx mice, the animal model of DMD. These engineered stem cells were fused with myoblasts and skeletal muscles of dystrophic scid/mdx mice, rescuing their dystrophin expression. We assessed that functional AONs were released in exosomes by these engineered, human DMD stem cells. These results demonstrated a bystander effect, mediated by the use of engineered stem cells for the delivery of AONs, and provided a possible new treatment method for DMDPubblicazioni consigliate
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