title: Evaluation of anhydrohexitol nucleic acid, cyclohexenyl nucleic acid and d-altritol nucleic acid-modified 2'-O-methyl RNA mixmer antisense oligonucleotides for exon skipping in vitro.
abstract: Antisense oligonucleotide (AO) mediated exon skipping has been widely explored as a therapeutic strategy for several diseases, in particular, for rare genetic disorders such as Duchenne muscular dystrophy (DMD). To date, the potential of anhydrohexitol nucleic acid (HNA), cyclohexenyl nucleic acid (CeNA) and altritol nucleic acid (ANA) has not been explored in exon skipping. For the first time, in this study we designed and synthesised HNA, CeNA and ANA-modified 2'-O-methyl (2'-OMe) mixmer AOs on a phosphorothioate (PS) backbone, and evaluated their potential to induce exon 23 skipping in mdx mouse myotubes, as a model system. Our results clearly showed that all three AO candidates modified with HNA, CeNA and ANA could efficiently induce Dmd exon 23 skipping in vitro in parallel to the fully modified 2'-OMePS AO with reduced dual exon 22/23 skipping. In addition, they showed high nuclease resistance and no cytotoxicity compared to the 2'-OMePS AO, demonstrating the applicability of HNA, CeNA and ANA nucleotide-modified AOs in exon skipping.
authors: Centre for Comparative Genomics, Murdoch University, Perth - 6150, Australia. R.Veedu@murdoch.edu.au and Western Australian Neuroscience Research Institute, Perth - 6150, Australia.
authors: Bao T
authors: BT
authors: Le
authors: Centre for Comparative Genomics, Murdoch University, Perth - 6150, Australia. R.Veedu@murdoch.edu.au and Western Australian Neuroscience Research Institute, Perth - 6150, Australia.
authors: Suxiang
authors: S
authors: Chen
authors: Laboratory for Medicinal Chemistry, Rega Institute for Medical Research, Katholieke Universiteit Leuven, Inderbroedersstraat 10, 3000 Leuven, Belgium.
authors: Mikhail
authors: M
authors: Abramov
authors: Laboratory for Medicinal Chemistry, Rega Institute for Medical Research, Katholieke Universiteit Leuven, Inderbroedersstraat 10, 3000 Leuven, Belgium.
authors: Piet
authors: P
authors: Herdewijn
authors: Centre for Comparative Genomics, Murdoch University, Perth - 6150, Australia. R.Veedu@murdoch.edu.au and Western Australian Neuroscience Research Institute, Perth - 6150, Australia.
authors: Rakesh N
authors: RN
authors: Veedu
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D053138:Cyclohexenes / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D005091:Exons / Q000235:genetics*; D051379:Mice; D018101:Mice, Inbred mdx; D032446:Myoblasts / Q000187:drug effects; D009696:Nucleic Acids / Q000737:chemistry*; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D012313:RNA / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013402:Sugar Alcohols / Q000737:chemistry*
chemical_list: D053138:Cyclohexenes; D009696:Nucleic Acids; D016376:Oligonucleotides, Antisense; D013402:Sugar Alcohols; C042890:altritol; D012313:RNA