{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T15:57:37Z","timestamp":1780329457184,"version":"3.54.1"},"reference-count":44,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2022,11,25]],"date-time":"2022-11-25T00:00:00Z","timestamp":1669334400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Grants-in-Aid for Scientific Research","doi-asserted-by":"publisher","award":["18\u00a0K07790"],"award-info":[{"award-number":["18\u00a0K07790"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["18ek0109249h0002"],"award-info":[{"award-number":["18ek0109249h0002"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["20ek0109405h0002"],"award-info":[{"award-number":["20ek0109405h0002"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["18ek0109318h0001"],"award-info":[{"award-number":["18ek0109318h0001"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,4,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>\u00a0<\/jats:title><jats:p>Fukuyama congenital muscular dystrophy (FCMD) is an autosomal recessive disorder caused by fukutin (FKTN) gene mutations. FCMD is the second most common form of childhood muscular dystrophy in Japan, and the most patients possess a homozygous retrotransposal SINE-VNTR-Alu insertion in the 3\u2032-untranslated region of FKTN. A deep-intronic variant (DIV) was previously identified as the second most prevalent loss-of-function mutation in Japanese patients with FCMD. The DIV creates a new splicing donor site in intron 5 that causes aberrant splicing and the formation of a 64-base pair pseudoexon in the mature mRNA, resulting in a truncated nonfunctional protein. Patients with FCMD carrying the DIV present a more severe symptoms, and currently, there is no radical therapy available for this disorder. In the present study, we describe in vitro evaluation of antisense oligonucleotide mediated skipping of pseudoexon inclusion and restoration of functional FKTN protein. A total of 16 19\u201326-mer antisense oligonucleotide sequences were designed with a 2\u2019-O-methyl backbone and were screened in patient-derived fibroblasts, lymphoblast cells and minigene splice assays. One antisense oligonucleotide targeting the exonic splice enhancer region significantly induced pseudoexon skipping and increased the expression of normal mRNA. It also rescued FKTN protein production in lymphoblast cells and restored functional O-mannosyl glycosylation of alpha-dystroglycan in patient-derived myotubes. Based on our results, antisense oligonucleotide-based splicing correction should be investigated further as a potential treatment for patients with FCMD carrying the DIV.<\/jats:p><\/jats:sec><jats:sec><jats:title>\u00a0<\/jats:title><jats:p>One Sentence Summary Antisense oligonucleotide treatment restored normal FKTN protein production and functional O-mannosyl glycosylation of alpha-dystroglycan via pseudoexon skipping in patient-derived cells carrying the compound heterozygous deep-intronic variant of Fukuyama muscular dystrophy.<\/jats:p><\/jats:sec>","DOI":"10.1093\/hmg\/ddac286","type":"journal-article","created":{"date-parts":[[2022,11,25]],"date-time":"2022-11-25T12:30:06Z","timestamp":1669379406000},"page":"1301-1312","source":"Crossref","is-referenced-by-count":9,"title":["Antisense oligonucleotide induced pseudoexon skipping and restoration of functional protein for Fukuyama muscular dystrophy caused by a deep-intronic variant"],"prefix":"10.1093","volume":"32","author":[{"given":"Sarantuya","family":"Enkhjargal","sequence":"first","affiliation":[{"name":"Fujita Health University Department of Molecular Genetics, Institute for Comprehensive Medical Science, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kana","family":"Sugahara","sequence":"additional","affiliation":[{"name":"Fujita Health University Hospital Department of Clinical Genetics, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1651-5549","authenticated-orcid":false,"given":"Behnoush","family":"Khaledian","sequence":"additional","affiliation":[{"name":"Fujita Health University Hospital Department of Clinical Genetics, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miwako","family":"Nagasaka","sequence":"additional","affiliation":[{"name":"Fujita Health University Hospital Department of Clinical Genetics, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidehito","family":"Inagaki","sequence":"additional","affiliation":[{"name":"Fujita Health University Department of Molecular Genetics, Institute for Comprehensive Medical Science, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5690-5218","authenticated-orcid":false,"given":"Hiroki","family":"Kurahashi","sequence":"additional","affiliation":[{"name":"Fujita Health University Department of Molecular Genetics, Institute for Comprehensive Medical Science, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8619-6678","authenticated-orcid":false,"given":"Hisatsugu","family":"Koshimizu","sequence":"additional","affiliation":[{"name":"Fujita Health University Research Promotion Headquarters, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatsushi","family":"Toda","sequence":"additional","affiliation":[{"name":"The University of Tokyo Department of Neurology, Graduate School of Medicine, , Bunkyo-ku, Tokyo 113-0033 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1607-400X","authenticated-orcid":false,"given":"Mariko","family":"Taniguchi-Ikeda","sequence":"additional","affiliation":[{"name":"Fujita Health University Hospital Department of Clinical Genetics, , Toyoake, Aichi 470-1192 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,11,25]]},"reference":[{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0387-7604(81)80002-2","article-title":"Congenital progressive muscular dystrophy of the Fukuyama type - clinical, genetic and pathological considerations","volume":"3","author":"Fukuyama","year":"1981","journal-title":"Brain Dev."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1016\/j.nmd.2018.08.001","article-title":"National registry of patients with Fukuyama congenital muscular dystrophy in Japan","volume":"28","author":"Ishigaki","year":"2018","journal-title":"Neuromuscul. Disord."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1177\/08830738050200041901","article-title":"Long-term prognosis of epilepsies and related seizure disorders in Fukuyama-type congenital muscular dystrophy","volume":"20","author":"Yoshioka","year":"2005","journal-title":"J. Child Neurol."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"e1187","DOI":"10.1542\/peds.2005-2469","article-title":"Cardiac involvement in Fukuyama-type congenital muscular dystrophy","volume":"117","author":"Nakanishi","year":"2006","journal-title":"Pediatrics"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.braindev.2007.05.012","article-title":"Seizure-genotype relationship in Fukuyama-type congenital muscular dystrophy","volume":"30","author":"Yoshioka","year":"2008","journal-title":"Brain Dev."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1016\/j.braindev.2017.05.008","article-title":"Cardiac involvement in Fukuyama muscular dystrophy is less severe than in Duchenne muscular dystrophy","volume":"39","author":"Yamamoto","year":"2017","journal-title":"Brain Dev."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.mam.2016.07.003","article-title":"Mechanistic aspects of the formation of \u03b1-dystroglycan and therapeutic research for the treatment of \u03b1-dystroglycanopathy: a review","volume":"51","author":"Taniguchi-Ikeda","year":"2016","journal-title":"Mol. Asp. Med."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1126\/science.1180512","article-title":"O-mannosyl phosphorylation of alpha-dystroglycan is required for laminin binding","volume":"327","author":"Yoshida-Moriguchi","year":"2010","journal-title":"Science"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"2209","DOI":"10.1016\/j.celrep.2016.02.017","article-title":"Identification of a post-translational modification with ribitol-phosphate and its defect in muscular dystrophy","volume":"14","author":"Kanagawa","year":"2016","journal-title":"Cell Rep."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1038\/28653","article-title":"An ancient retrotransposal insertion causes Fukuyama-type congenital muscular dystrophy","volume":"394","author":"Kobayashi","year":"1998","journal-title":"Nature"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1007\/s004390000421","article-title":"Age and origin of the FCMD 3\u2032-untranslated-region retrotransposal insertion mutation causing Fukuyama-type congenital muscular dystrophy in the Japanese population","volume":"107","author":"Colombo","year":"2000","journal-title":"Hum. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1038\/nature10456","article-title":"Pathogenic exon-trapping by SVA retrotransposon and rescue in Fukuyama muscular dystrophy","volume":"478","author":"Taniguchi-Ikeda","year":"2011","journal-title":"Nature"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1016\/j.nmd.2010.06.005","article-title":"Fukutin mutations in congenital muscular dystrophies with defective glycosylation of dystroglycan in Korea","volume":"20","author":"Lim","year":"2010","journal-title":"Neuromuscul. Disord."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1038\/jhg.2017.71","article-title":"Deep-intronic variant of fukutin is the most prevalent point mutation of Fukuyama congenital muscular dystrophy in Japan","volume":"62","author":"Kobayashi","year":"2017","journal-title":"J. Hum. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1016\/j.jiac.2019.12.015","article-title":"Acute rhabdomyolysis following viral infection with coxsackie A4 in a 50-day-old infant with Fukuyama congenital muscular dystrophy","volume":"26","author":"Yamaguchi","year":"2020","journal-title":"J. Infect. Chemother."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1038\/jhg.2010.49","article-title":"Mutation spectrum of the dystrophin gene in 442 Duchenne\/Becker muscular dystrophy cases from one Japanese referral center","volume":"55","author":"Takeshima","year":"2010","journal-title":"J. Hum. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1093\/hmg\/7.3.393","article-title":"A G--&gt;a transition creates a branch point sequence and activation of a cryptic exon, resulting in the hereditary disorder neurofibromatosis 2","volume":"7","author":"De Klein","year":"1998","journal-title":"Hum. Mol. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1007\/s00439-017-1809-4","article-title":"Deep intronic mutations and human disease","volume":"136","author":"Vaz-Drago","year":"2017","journal-title":"Hum. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1900","DOI":"10.1016\/j.febslet.2005.02.047","article-title":"Are splicing mutations the most frequent cause of hereditary disease?","volume":"579","author":"L\u00f3pez-Bigas","year":"2005","journal-title":"FEBS Lett."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1089\/hum.2015.070","article-title":"Oligonucleotide therapies: the past and the present","volume":"26","author":"Lundin","year":"2015","journal-title":"Hum. Gene Ther."},{"key":"2023040610573162600_","first-page":"347","article-title":"Antisense oligonucleotides: basic concepts and mechanisms","volume":"1","author":"Dias","year":"2002","journal-title":"Mol. Cancer Ther."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1002\/humu.20839","article-title":"A deep intronic mutation in FGB creates a consensus exonic splicing enhancer motif that results in afibrinogenemia caused by aberrant mRNA splicing, which can be corrected in vitro with antisense oligonucleotide treatment","volume":"30","author":"Davis","year":"2009","journal-title":"Hum. Mutat."},{"key":"2023040610573162600_","first-page":"2552","article-title":"In vitro and in vivo rescue of aberrant splicing in CEP290-associated LCA by antisense oligonucleotide delivery","volume":"25","author":"Garanto","year":"2016","journal-title":"Hum. Mol. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1751","DOI":"10.1038\/s41436-018-0414-9","article-title":"Deep-intronic ABCA4 variants explain missing heritability in Stargardt disease and allow correction of splice defects by antisense oligonucleotides","volume":"21","author":"Sangermano","year":"2019","journal-title":"Genet. Med."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1016\/j.omtn.2020.07.036","article-title":"Antisense oligonucleotide- and CRISPR-Cas9-mediated rescue of mRNA splicing for a deep intronic CLRN1 mutation","volume":"21","author":"Panagiotopoulos","year":"2020","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1089\/nat.2017.0665","article-title":"FDA approval of nusinersen for spinal muscular atrophy makes 2016 the year of splice modulating oligonucleotides","volume":"27","author":"Aartsma-Rus","year":"2017","journal-title":"Nucleic Acid Ther."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1056\/NEJMoa1716793","article-title":"Inotersen treatment for patients with hereditary transthyretin amyloidosis","volume":"379","author":"Benson","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1358\/dot.2019.55.10.3045038","article-title":"Viltolarsen for the treatment of Duchenne muscular dystrophy","volume":"55","author":"Roshmi","year":"2019","journal-title":"Drugs Today"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1007\/s40265-020-01339-3","article-title":"Viltolarsen: first approval","volume":"80","author":"Dhillon","year":"2020","journal-title":"Drugs"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1038\/sj.gt.3302313","article-title":"Comparative analysis of antisense oligonucleotide analogs for targeted DMD exon 46 skipping in muscle cells","volume":"11","author":"Aartsma-Rus","year":"2004","journal-title":"Gene Ther."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"103140","DOI":"10.1016\/j.isci.2021.103140","article-title":"Restoration of the defect in radial glial fiber migration and cortical plate organization in a brain organoid model of Fukuyama muscular dystrophy","volume":"24","author":"Taniguchi-Ikeda","year":"2021","journal-title":"iScience"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1111\/j.1742-4658.2009.07520.x","article-title":"Alternative splicing: role of pseudoexons in human disease and potential therapeutic strategies","volume":"277","author":"Dhir","year":"2010","journal-title":"FEBS J."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1038\/gim.2013.176","article-title":"In silico tools for splicing defect prediction: a survey from the viewpoint of end users","volume":"16","author":"Jian","year":"2014","journal-title":"Genet. Med."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1038\/nature22799","article-title":"Structure of a pre-catalytic spliceosome","volume":"546","author":"Plaschka","year":"2017","journal-title":"Nature"},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/nrg775","article-title":"Listening to silence and understanding nonsense: exonic mutations that affect splicing","volume":"3","author":"Cartegni","year":"2002","journal-title":"Nat. Rev. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/S0968-0004(00)01549-8","article-title":"Exonic splicing enhancers: mechanism of action, diversity and role in human genetic diseases","volume":"25","author":"Blencowe","year":"2000","journal-title":"Trends Biochem. Sci."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1042\/BJ20081501","article-title":"The SR protein family of splicing factors: master regulators of gene expression","volume":"417","author":"Long","year":"2009","journal-title":"Biochem. J."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1007\/s00439-016-1683-5","article-title":"The hnRNP family: insights into their role in health and disease","volume":"135","author":"Geuens","year":"2016","journal-title":"Hum. Genet."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"6549","DOI":"10.1093\/nar\/gkw533","article-title":"Splice-switching antisense oligonucleotides as therapeutic drugs","volume":"44","author":"Havens","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"8673","DOI":"10.1073\/pnas.90.18.8673","article-title":"Restoration of correct splicing in thalassemic pre-mRNA by antisense oligonucleotides","volume":"90","author":"Dominski","year":"1993","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1644","DOI":"10.1056\/NEJMoa1813279","article-title":"Patient-customized oligonucleotide therapy for a rare genetic disease","volume":"381","author":"Kim","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1038\/mt.2008.205","article-title":"Guidelines for antisense oligonucleotide design and insight into splice-modulating mechanisms","volume":"17","author":"Aartsma-Rus","year":"2009","journal-title":"Mol. Ther."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"1387","DOI":"10.1002\/humu.22379","article-title":"In vitro correction of a pseudoexon-generating deep intronic mutation in LGMD2A by antisense oligonucleotides and modified small nuclear RNAs","volume":"34","author":"Blazquez","year":"2013","journal-title":"Hum. Mutat."},{"key":"2023040610573162600_","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.omtn.2020.05.029","article-title":"Exon-skipping oligonucleotides restore functional collagen VI by correcting a common COL6A1 mutation in Ullrich CMD","volume":"21","author":"Aguti","year":"2020","journal-title":"Mol. Ther. Nucleic Acids"}],"container-title":["Human Molecular Genetics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/hmg\/advance-article-pdf\/doi\/10.1093\/hmg\/ddac286\/48473252\/ddac286.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/hmg\/article-pdf\/32\/8\/1301\/49778374\/ddac286.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/hmg\/article-pdf\/32\/8\/1301\/49778374\/ddac286.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,9]],"date-time":"2024-10-09T12:06:40Z","timestamp":1728475600000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/hmg\/article\/32\/8\/1301\/6847082"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,25]]},"references-count":44,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,11,25]]},"published-print":{"date-parts":[[2023,4,6]]}},"URL":"https:\/\/doi.org\/10.1093\/hmg\/ddac286","relation":{},"ISSN":["0964-6906","1460-2083"],"issn-type":[{"value":"0964-6906","type":"print"},{"value":"1460-2083","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2023,4,15]]},"published":{"date-parts":[[2022,11,25]]}}}