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Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology

Glycogen storage disease type II (GSDII) is a lysosomal disorder caused by the deficient activity of acid alpha-glucosidase (GAA) enzyme, leading to the accumulation of glycogen within the lysosomes. The disease has been classified in infantile and late-onset forms. Most late-onset patients share a...

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Autores principales: Goina, Elisa, Peruzzo, Paolo, Bembi, Bruno, Dardis, Andrea, Buratti, Emanuele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589062/
https://www.ncbi.nlm.nih.gov/pubmed/28629821
http://dx.doi.org/10.1016/j.ymthe.2017.05.019
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author Goina, Elisa
Peruzzo, Paolo
Bembi, Bruno
Dardis, Andrea
Buratti, Emanuele
author_facet Goina, Elisa
Peruzzo, Paolo
Bembi, Bruno
Dardis, Andrea
Buratti, Emanuele
author_sort Goina, Elisa
collection PubMed
description Glycogen storage disease type II (GSDII) is a lysosomal disorder caused by the deficient activity of acid alpha-glucosidase (GAA) enzyme, leading to the accumulation of glycogen within the lysosomes. The disease has been classified in infantile and late-onset forms. Most late-onset patients share a splicing mutation c.-32-13T > G in intron 1 of the GAA gene that prevents efficient recognition of exon 2 by the spliceosome. In this study, we have mapped the splicing silencers of GAA exon 2 and developed antisense morpholino oligonucleotides (AMOs) to inhibit those regions and rescue normal splicing in the presence of the c.-32-13T > G mutation. Using a minigene approach and patient fibroblasts, we successfully increased inclusion of exon 2 in the mRNA and GAA enzyme production by targeting a specific silencer with a combination of AMOs. Most importantly, the use of these AMOs in patient myotubes results in a decreased accumulation of glycogen. To our knowledge, this is the only therapeutic approach resulting in a decrease of glycogen accumulation in patient tissues beside enzyme replacement therapy (ERT) and TFEB overexpression. As a result, it may represent a highly novel and promising therapeutic line for GSDII.
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spelling pubmed-55890622018-09-06 Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology Goina, Elisa Peruzzo, Paolo Bembi, Bruno Dardis, Andrea Buratti, Emanuele Mol Ther Original Article Glycogen storage disease type II (GSDII) is a lysosomal disorder caused by the deficient activity of acid alpha-glucosidase (GAA) enzyme, leading to the accumulation of glycogen within the lysosomes. The disease has been classified in infantile and late-onset forms. Most late-onset patients share a splicing mutation c.-32-13T > G in intron 1 of the GAA gene that prevents efficient recognition of exon 2 by the spliceosome. In this study, we have mapped the splicing silencers of GAA exon 2 and developed antisense morpholino oligonucleotides (AMOs) to inhibit those regions and rescue normal splicing in the presence of the c.-32-13T > G mutation. Using a minigene approach and patient fibroblasts, we successfully increased inclusion of exon 2 in the mRNA and GAA enzyme production by targeting a specific silencer with a combination of AMOs. Most importantly, the use of these AMOs in patient myotubes results in a decreased accumulation of glycogen. To our knowledge, this is the only therapeutic approach resulting in a decrease of glycogen accumulation in patient tissues beside enzyme replacement therapy (ERT) and TFEB overexpression. As a result, it may represent a highly novel and promising therapeutic line for GSDII. American Society of Gene & Cell Therapy 2017-09-06 2017-06-16 /pmc/articles/PMC5589062/ /pubmed/28629821 http://dx.doi.org/10.1016/j.ymthe.2017.05.019 Text en © 2017 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Goina, Elisa
Peruzzo, Paolo
Bembi, Bruno
Dardis, Andrea
Buratti, Emanuele
Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology
title Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology
title_full Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology
title_fullStr Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology
title_full_unstemmed Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology
title_short Glycogen Reduction in Myotubes of Late-Onset Pompe Disease Patients Using Antisense Technology
title_sort glycogen reduction in myotubes of late-onset pompe disease patients using antisense technology
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589062/
https://www.ncbi.nlm.nih.gov/pubmed/28629821
http://dx.doi.org/10.1016/j.ymthe.2017.05.019
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