Pseudoexon exclusion by antisense therapy in methylmalonic aciduria (MMAuria)†
B. Pérez
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
B. Pérez and A. Rincón contributed equally to this work.
Search for more papers by this authorA. Rincón
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
B. Pérez and A. Rincón contributed equally to this work.
Search for more papers by this authorA. Jorge-Finnigan
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorE. Richard
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorB. Merinero
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorCorresponding Author
M. Ugarte
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Centro de Biología Molecular “Severo Ochoa” CSIC-UAM, C/Nicolás Cabrera No. 1, Universidad Autónoma de Madrid, 28049 Madrid, SpainSearch for more papers by this authorL.R. Desviat
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorB. Pérez
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
B. Pérez and A. Rincón contributed equally to this work.
Search for more papers by this authorA. Rincón
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
B. Pérez and A. Rincón contributed equally to this work.
Search for more papers by this authorA. Jorge-Finnigan
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorE. Richard
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorB. Merinero
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorCorresponding Author
M. Ugarte
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Centro de Biología Molecular “Severo Ochoa” CSIC-UAM, C/Nicolás Cabrera No. 1, Universidad Autónoma de Madrid, 28049 Madrid, SpainSearch for more papers by this authorL.R. Desviat
Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular “Severo Ochoa” Universidad Autónoma de Madrid (UAM)–Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid, Madrid, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Madrid, Spain
Search for more papers by this authorCommunicated by David S. Rosenblatt
Abstract
Development of pseudoexon exclusion therapies by antisense modification of pre-mRNA splicing represents a type of personalized genetic medicine. Here we present the cellular antisense therapy and the cell-based splicing assays to investigate the effect of two novel deep intronic changes c.1957–898A>G and c.1957–920C>A identified in the methylmalonyl–coenzyme A (CoA) mutase (MUT) gene. The results show that the nucleotide change c.1957–898A>G is a pathological mutation activating pseudoexon insertion and that antisense morpholino oligonucleotide (AMO) treatment in patient fibroblasts leads to recovery of MUT activity to levels 25 to 100% of control range. On the contrary, the change c.1957–920C>A, identified in two fibroblasts cell lines in cis with c.1885A>G (p.R629G) or c.458T>A (p.D153V), appears to be a rare variant of uncertain clinical significance. The functional analysis of c.1885A>G and c.458T>A indicate that they are the disease-causing mutations in these two patients. The results presented here highlight the necessity of scanning the described intronic region for mutations in MUT-affected patients, followed by functional analyses to demonstrate the pathogenicity of the identified changes, and extend previous work of the applicability of the antisense approach in methylmalonic aciduria (MMAuria) for a novel intronic mutation. Hum Mutat 30:1–7, 2009. © 2009 Wiley-Liss, Inc.
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