title: Assessment of Targeted Next-Generation Sequencing as a Tool for the Diagnosis of Charcot-Marie-Tooth Disease and Hereditary Motor Neuropathy.
abstract: Charcot-Marie-Tooth disease is characterized by broad genetic heterogeneity with >50 known disease-associated genes. Mutations in some of these genes can cause a pure motor form of hereditary motor neuropathy, the genetics of which are poorly characterized. We designed a panel comprising 56 genes associated with Charcot-Marie-Tooth disease/hereditary motor neuropathy. We validated this diagnostic tool by first testing 11 patients with pathological mutations. A cohort of 33 affected subjects was selected for this study. The DNAJB2 c.352+1G>A mutation was detected in two cases; novel changes and/or variants with low frequency (<1%) were found in 12 cases. There were no candidate variants in 18 cases, and amplification failed for one sample. The DNAJB2 c.352+1G>A mutation was also detected in three additional families. On haplotype analysis, all of the patients from these five families shared the same haplotype; therefore, the DNAJB2 c.352+1G>A mutation may be a founder event. Our gene panel allowed us to perform a very rapid and cost-effective screening of genes involved in Charcot-Marie-Tooth disease/hereditary motor neuropathy. Our diagnostic strategy was robust in terms of both coverage and read depth for all of the genes and patient samples. These findings demonstrate the difficulty in achieving a definitive molecular diagnosis because of the complexity of interpreting new variants and the genetic heterogeneity that is associated with these neuropathies.
authors.affiliation: Unit of Genetics and Genomics of Neuromuscular Disorders, Program in Rare and Genetic Diseases and IBV/CSIC Associated Unit, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain; Unit 732, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Valencia, Spain; Department of Genomics and Translational Genetics, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Lupo
authors.affiliation: Unit 715, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Valencia, Spain; Unit of Systems Biology, Program in Computational Genomics, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: García-García
authors.affiliation: Unit of Genetics and Genomics of Neuromuscular Disorders, Program in Rare and Genetic Diseases and IBV/CSIC Associated Unit, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain; Unit 732, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Valencia, Spain.
authors.forename: Paula
authors.initials: P
authors.lastname: Sancho
authors.affiliation: Unit of Genetics and Genomics of Neuromuscular Disorders, Program in Rare and Genetic Diseases and IBV/CSIC Associated Unit, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Tello
authors.affiliation: Department of Neuropediatrics, Hospital Universitario La Paz, Madrid, Spain.
authors.forename: Mar
authors.initials: M
authors.lastname: García-Romero
authors.affiliation: Department of Neurology, Hospital Universitario Virgen del Rocío, Seville, Spain.
authors.forename: Liliana
authors.initials: L
authors.lastname: Villarreal
authors.affiliation: Department of Neurology, Hospital de Bellvitge, Barcelona, Spain.
authors.forename: Antonia
authors.initials: A
authors.lastname: Alberti
authors.affiliation: Department of Neurology, Hospital Universitari i Politècnic La Fe and Instituto de Investigación Sanitario (IIS) - La Fe, Valencia, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Sivera
authors.affiliation: Unit 715, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Valencia, Spain; Unit of Systems Biology, Program in Computational Genomics, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain; Functional Genomics Node, Spanish National Institute of Bioinformatics (INB), Valencia, Spain.
authors.forename: Joaquín
authors.initials: J
authors.lastname: Dopazo
authors.affiliation: Unit of Systems Biology, Program in Computational Genomics, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain.
authors.forename: Samuel I
authors.initials: SI
authors.lastname: Pascual-Pascual
authors.affiliation: Department of Neuropediatrics, Hospital Universitario La Paz, Madrid, Spain.
authors.forename: Celedonio
authors.initials: C
authors.lastname: Márquez-Infante
authors.affiliation: Department of Neurology, Hospital de Bellvitge, Barcelona, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Casasnovas
authors.affiliation: Department of Neurology, Hospital Universitari i Politècnic La Fe and Instituto de Investigación Sanitario (IIS) - La Fe, Valencia, Spain; Unit 763, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Valencia, Spain; Department of Medicine, Universitat de València, Valencia, Spain.
authors.forename: Teresa
authors.initials: T
authors.lastname: Sevilla
authors.affiliation: Unit of Genetics and Genomics of Neuromuscular Disorders, Program in Rare and Genetic Diseases and IBV/CSIC Associated Unit, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain; Unit 732, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Valencia, Spain; Department of Genomics and Translational Genetics, Centro de Investigación Príncipe Felipe (CIPF), Valencia, Spain. Electronic address: cespinos@cipf.es.
authors.forename: Carmen
authors.initials: C
authors.lastname: Espinós
mesh_terms: D016022:Case-Control Studies; D002607:Charcot-Marie-Tooth Disease / Q000175:diagnosis* / Q000235:genetics*; D005260:Female; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics; D006239:Haplotypes; D015417:Hereditary Sensory and Motor Neuropathy / Q000175:diagnosis* / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D018832:Molecular Chaperones / Q000235:genetics; D009154:Mutation; D015203:Reproducibility of Results
chemical_list: C121958:DNAJB2 protein, human; D050956:HSP40 Heat-Shock Proteins; D018832:Molecular Chaperones