Triplet-repeat oligonucleotide-mediated reversal of RNA toxicity in myotonic dystrophy.
Myotonic dystrophy type 1 (DM1) is caused by toxicity of an expanded, noncoding (CUG)n tract in DM protein kinase (DMPK) transcripts. According to current evidence the long (CUG)n segment is involved in entrapment of muscleblind (Mbnl) proteins in ribonuclear aggregates and stabilized expression of CUG binding protein 1 (CUGBP1), causing aberrant premRNA splicing and associated pathogenesis in DM1 patients. Here, we report on the use of antisense oligonucleotides (AONs) in a therapeutic strategy for reversal of RNA-gain-of-function toxicity. Using a previously undescribed mouse DM1 myoblast-myotube cell model and DM1 patient cells as screening tools, we have identified a fully 2'-O-methyl-phosphorothioate-modified (CAG)7 AON that silences mutant DMPK RNA expression and reduces the number of ribonuclear aggregates in a selective and (CUG)n-length-dependent manner. Direct administration of this AON in muscle of DM1 mouse models in vivo caused a significant reduction in the level of toxic (CUG)n RNA and a normalizing effect on aberrant premRNA splicing. Our data demonstrate proof of principle for therapeutic use of simple sequence AONs in DM1 and potentially other unstable microsatellite diseases.
Mulders
Susan A M
SA
Department of Cell Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
van den Broek
Walther J A A
WJ
Wheeler
Thurman M
TM
Croes
Huib J E
HJ
van Kuik-Romeijn
Petra
P
de Kimpe
Sjef J
SJ
Furling
Denis
D
Platenburg
Gerard J
GJ
Gourdon
Geneviève
G
Thornton
Charles A
CA
Wieringa
Bé
B
Wansink
Derick G
DG
D000483:Alleles; D000818:Animals; D000067879:CELF1 Protein; D020868:Gene Silencing; D051379:Mice; D008957:Models, Genetic; D018482:Muscle, Skeletal / Q000378:metabolism; D009154:Mutation; D032446:Myoblasts / Q000378:metabolism; D009223:Myotonic Dystrophy / Q000235:genetics* / Q000628:therapy; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics*; D016376:Oligonucleotides, Antisense / Q000235:genetics; D012313:RNA / Q000235:genetics*; D034622:RNA Interference; D012326:RNA Splicing; D016601:RNA-Binding Proteins / Q000235:genetics
D000067879:CELF1 Protein; C502743:CELF1 protein, mouse; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D016601:RNA-Binding Proteins; D012313:RNA