GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells
Pompe disease is a metabolic myopathy caused by deficiency of the acid α-glucosidase (GAA) enzyme and results in progressive wasting of skeletal muscle cells. The c.-32-13T>G (IVS1) GAA variant promotes exon 2 skipping during pre-mRNA splicing and is the most common variant for the childhood/adul...
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doaj-9f9c7d67ca804d6bbf6aabc1dbc487b82020-11-24T23:11:57ZengElsevierMolecular Therapy: Nucleic Acids2162-25312017-06-017C10111510.1016/j.omtn.2017.03.002GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle CellsErik van der Wal0Atze J. Bergsma1Tom J.M. van Gestel2Stijn L.M. in ‘t Groen3Holm Zaehres4Marcos J. Araúzo-Bravo5Hans R. Schöler6Ans T. van der Ploeg7W.W.M. Pim Pijnappel8Molecular Stem Cell Biology, Department of Clinical Genetics, Erasmus Medical Center, 3015 CN Rotterdam, the NetherlandsMolecular Stem Cell Biology, Department of Clinical Genetics, Erasmus Medical Center, 3015 CN Rotterdam, the NetherlandsMolecular Stem Cell Biology, Department of Clinical Genetics, Erasmus Medical Center, 3015 CN Rotterdam, the NetherlandsMolecular Stem Cell Biology, Department of Clinical Genetics, Erasmus Medical Center, 3015 CN Rotterdam, the NetherlandsDepartment of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, GermanyDepartment of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, GermanyDepartment of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, 48149 Münster, GermanyDepartment of Pediatrics, Erasmus Medical Center, 3015 CN Rotterdam, the NetherlandsMolecular Stem Cell Biology, Department of Clinical Genetics, Erasmus Medical Center, 3015 CN Rotterdam, the NetherlandsPompe disease is a metabolic myopathy caused by deficiency of the acid α-glucosidase (GAA) enzyme and results in progressive wasting of skeletal muscle cells. The c.-32-13T>G (IVS1) GAA variant promotes exon 2 skipping during pre-mRNA splicing and is the most common variant for the childhood/adult disease form. We previously identified antisense oligonucleotides (AONs) that promoted GAA exon 2 inclusion in patient-derived fibroblasts. It was unknown how these AONs would affect GAA splicing in skeletal muscle cells. To test this, we expanded induced pluripotent stem cell (iPSC)-derived myogenic progenitors and differentiated these to multinucleated myotubes. AONs restored splicing in myotubes to a similar extent as in fibroblasts, suggesting that they act by modulating the action of shared splicing regulators. AONs targeted the putative polypyrimidine tract of a cryptic splice acceptor site that was part of a pseudo exon in GAA intron 1. Blocking of the cryptic splice donor of the pseudo exon with AONs likewise promoted GAA exon 2 inclusion. The simultaneous blocking of the cryptic acceptor and cryptic donor sites restored the majority of canonical splicing and alleviated GAA enzyme deficiency. These results highlight the relevance of cryptic splicing in human disease and its potential as therapeutic target for splicing modulation using AONs.http://www.sciencedirect.com/science/article/pii/S2162253117301403myogenic progenitorsinduced pluripotent stem cellantisense oligonucleotidePompe diseaseIVS1 mutationskeletal muscle differentiationpre-mRNA splicingpseudo exonexon inclusionlysosomal storage disorder |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Erik van der Wal Atze J. Bergsma Tom J.M. van Gestel Stijn L.M. in ‘t Groen Holm Zaehres Marcos J. Araúzo-Bravo Hans R. Schöler Ans T. van der Ploeg W.W.M. Pim Pijnappel |
spellingShingle |
Erik van der Wal Atze J. Bergsma Tom J.M. van Gestel Stijn L.M. in ‘t Groen Holm Zaehres Marcos J. Araúzo-Bravo Hans R. Schöler Ans T. van der Ploeg W.W.M. Pim Pijnappel GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells Molecular Therapy: Nucleic Acids myogenic progenitors induced pluripotent stem cell antisense oligonucleotide Pompe disease IVS1 mutation skeletal muscle differentiation pre-mRNA splicing pseudo exon exon inclusion lysosomal storage disorder |
author_facet |
Erik van der Wal Atze J. Bergsma Tom J.M. van Gestel Stijn L.M. in ‘t Groen Holm Zaehres Marcos J. Araúzo-Bravo Hans R. Schöler Ans T. van der Ploeg W.W.M. Pim Pijnappel |
author_sort |
Erik van der Wal |
title |
GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells |
title_short |
GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells |
title_full |
GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells |
title_fullStr |
GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells |
title_full_unstemmed |
GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells |
title_sort |
gaa deficiency in pompe disease is alleviated by exon inclusion in ipsc-derived skeletal muscle cells |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2017-06-01 |
description |
Pompe disease is a metabolic myopathy caused by deficiency of the acid α-glucosidase (GAA) enzyme and results in progressive wasting of skeletal muscle cells. The c.-32-13T>G (IVS1) GAA variant promotes exon 2 skipping during pre-mRNA splicing and is the most common variant for the childhood/adult disease form. We previously identified antisense oligonucleotides (AONs) that promoted GAA exon 2 inclusion in patient-derived fibroblasts. It was unknown how these AONs would affect GAA splicing in skeletal muscle cells. To test this, we expanded induced pluripotent stem cell (iPSC)-derived myogenic progenitors and differentiated these to multinucleated myotubes. AONs restored splicing in myotubes to a similar extent as in fibroblasts, suggesting that they act by modulating the action of shared splicing regulators. AONs targeted the putative polypyrimidine tract of a cryptic splice acceptor site that was part of a pseudo exon in GAA intron 1. Blocking of the cryptic splice donor of the pseudo exon with AONs likewise promoted GAA exon 2 inclusion. The simultaneous blocking of the cryptic acceptor and cryptic donor sites restored the majority of canonical splicing and alleviated GAA enzyme deficiency. These results highlight the relevance of cryptic splicing in human disease and its potential as therapeutic target for splicing modulation using AONs. |
topic |
myogenic progenitors induced pluripotent stem cell antisense oligonucleotide Pompe disease IVS1 mutation skeletal muscle differentiation pre-mRNA splicing pseudo exon exon inclusion lysosomal storage disorder |
url |
http://www.sciencedirect.com/science/article/pii/S2162253117301403 |
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