title: Short antisense-locked nucleic acids (all-LNAs) correct alternative splicing abnormalities in myotonic dystrophy.
abstract: Myotonic dystrophy type 1 (DM1) is an autosomal dominant multisystemic disorder caused by expansion of CTG triplet repeats in 3'-untranslated region of DMPK gene. The pathomechanism of DM1 is driven by accumulation of toxic transcripts containing expanded CUG repeats (CUG(exp)) in nuclear foci which sequester several factors regulating RNA metabolism, such as Muscleblind-like proteins (MBNLs). In this work, we utilized very short chemically modified antisense oligonucleotides composed exclusively of locked nucleic acids (all-LNAs) complementary to CUG repeats, as potential therapeutic agents against DM1. Our in vitro data demonstrated that very short, 8- or 10-unit all-LNAs effectively bound the CUG repeat RNA and prevented the formation of CUG(exp)/MBNL complexes. In proliferating DM1 cells as well as in skeletal muscles of DM1 mouse model the all-LNAs induced the reduction of the number and size of CUG(exp) foci and corrected MBNL-sensitive alternative splicing defects with high efficacy and specificity. The all-LNAs had low impact on the cellular level of CUG(exp)-containing transcripts and did not affect the expression of other transcripts with short CUG repeats. Our data strongly indicate that short all-LNAs complementary to CUG repeats are a promising therapeutic tool against DM1.
authors.affiliation: Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Wojtkowiak-Szlachcic
authors.affiliation: Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Taylor
authors.affiliation: Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Stepniak-Konieczna
authors.affiliation: Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
authors.forename: Lukasz J
authors.initials: LJ
authors.lastname: Sznajder
authors.affiliation: Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Mykowska
authors.affiliation: Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Sroka
authors.affiliation: Department of Neurology, Box 645, University of Rochester Medical Center, 601 Elmwood Ave, Rochester, NY 14642, USA.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Thornton
authors.affiliation: Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland ksobczak@amu.edu.pl.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Sobczak
mesh_terms: D020413:3' Untranslated Regions; D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004195:Disease Models, Animal; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D050505:Mutant Proteins / Q000235:genetics; D009223:Myotonic Dystrophy / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D065636:Myotonin-Protein Kinase / Q000235:genetics; D009841:Oligonucleotides / Q000235:genetics / Q000627:therapeutic use*; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000627:therapeutic use*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D019680:Trinucleotide Repeat Expansion
chemical_list: D020413:3' Untranslated Regions; C585517:DMPK protein, human; C493663:MBNL1 protein, human; D050505:Mutant Proteins; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D016601:RNA-Binding Proteins; C477371:locked nucleic acid; D065636:Myotonin-Protein Kinase