title: Next-generation sequencing of 28 ALS-related genes in a Japanese ALS cohort.
abstract: We investigated the frequency and contribution of variants of the 28 known amyotrophic lateral sclerosis (ALS)-related genes in Japanese ALS patients. We designed a multiplex, polymerase chain reaction-based primer panel to amplify the coding regions of the 28 ALS-related genes and sequenced DNA samples from 257 Japanese ALS patients using an Ion Torrent PGM sequencer. We also performed exome sequencing and identified variants of the 28 genes in an additional 251 ALS patients using an Illumina HiSeq 2000 platform. We identified the known ALS pathogenic variants and predicted the functional properties of novel nonsynonymous variants in silico. These variants were confirmed by Sanger sequencing. Known pathogenic variants were identified in 19 (48.7%) of the 39 familial ALS patients and 14 (3.0%) of the 469 sporadic ALS patients. Thirty-two sporadic ALS patients (6.8%) harbored 1 or 2 novel nonsynonymous variants of ALS-related genes that might be deleterious. This study reports the first extensive genetic screening of Japanese ALS patients. These findings are useful for developing genetic screening and counseling strategies for such patients.
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Ryoichi
authors.initials: R
authors.lastname: Nakamura
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Sone
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Atsuta
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Genki
authors.initials: G
authors.lastname: Tohnai
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Department of Neurology, Japanese Red Cross Nagoya Daiichi Hospital, Nagoya, Japan.
authors.forename: Hazuki
authors.initials: H
authors.lastname: Watanabe
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Daichi
authors.initials: D
authors.lastname: Yokoi
authors.affiliation: Bioinformatics Section, Center for Advanced Medicine and Clinical Research, Nagoya University Hospital, Nagoya, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nakatochi
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Brain and Mind Research Center, Nagoya University, Nagoya, Japan.
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Watanabe
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Mizuki
authors.initials: M
authors.lastname: Ito
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Department of Neurology, Komaki City Hospital, Komaki, Japan.
authors.forename: Jo
authors.initials: J
authors.lastname: Senda
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Masahisa
authors.initials: M
authors.lastname: Katsuno
authors.affiliation: Department of Neurology and Stroke Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Tanaka
authors.affiliation: Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Yuanzhe
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Neurology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima, Japan.
authors.forename: Yuishin
authors.initials: Y
authors.lastname: Izumi
authors.affiliation: Division of Neurology, Department of Internal Medicine, Jichi Medical University, Tochigi, Japan.
authors.forename: Mitsuya
authors.initials: M
authors.lastname: Morita
authors.affiliation: Department of Neurology, Mie University Graduate School of Medicine, Tsu, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Taniguchi
authors.affiliation: Division of Neurology, Department of Internal Medicine, Toho University School of Medicine, Tokyo, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Kano
authors.affiliation: Department of Neurology, Vihara Hananosato Hospital, Miyoshi, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Oda
authors.affiliation: Department of Neurology, Graduate School of Medicine, Chiba University, Chiba, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kuwabara
authors.affiliation: Department of Neurology, Okayama University Graduate School of Medicine, Okayama, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Abe
authors.affiliation: Department of Neurology, National Hospital Organization Higashinagoya National Hospital, Nagoya, Japan.
authors.forename: Ikuko
authors.initials: I
authors.lastname: Aiba
authors.affiliation: Department of Neurology, Geriatrics Research Institute and Hospital, Maebashi, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Okamoto
authors.affiliation: Department of Neurology, National Hospital Organization Shizuoka-Fuji Hospital, Fujinomiya, Japan.
authors.forename: Kouichi
authors.initials: K
authors.lastname: Mizoguchi
authors.affiliation: Division of Neurology, National Hospital Organization, Sagamihara National Hospital, Sagamihara, Japan.
authors.forename: Kazuko
authors.initials: K
authors.lastname: Hasegawa
authors.affiliation: Department of Neurology, Tohoku University School of Medicine, Sendai, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Aoki
authors.affiliation: Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Hattori
authors.affiliation: Department of Neurology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Shoji
authors.initials: S
authors.lastname: Tsuji
authors.affiliation: Division of Neurology, Department of Brain and Neurosciences, Faculty of Medicine, Tottori University, Yonago, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Nakashima
authors.affiliation: Department of Neurology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Kaji
authors.affiliation: Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan; Research Division of Dementia and Neurodegenerative Disease, Nagoya University Graduate School of Medicine, Nagoya, Japan. Electronic address: sobueg@med.nagoya-u.ac.jp.
authors.forename: Gen
authors.initials: G
authors.lastname: Sobue
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics*; D044466:Asian People; D000073885:C9orf72 Protein; D015331:Cohort Studies; D004247:DNA / Q000235:genetics*; D059472:Exome / Q000235:genetics; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics*; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D011506:Proteins / Q000235:genetics; D034702:RNA-Binding Protein FUS / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods* / Q000639:trends*; D000072105:Superoxide Dismutase-1 / Q000235:genetics
chemical_list: C438187:ALS2 protein, human; D000073885:C9orf72 Protein; C568651:C9orf72 protein, human; C000599550:FUS protein, human; D020662:Guanine Nucleotide Exchange Factors; D011506:Proteins; D034702:RNA-Binding Protein FUS; C000606290:SOD1 protein, human; C520412:SPG11 protein, human; D004247:DNA; D000072105:Superoxide Dismutase-1
keywords: Amyotrophic lateral sclerosis; Genetic screening; Japan; Next-generation sequencer