Development of molecular therapy for epilepsy using knock-in rats and artificial patient iPS cells with next generation methodologies

2018 Fiscal Year Annual Research Report

• Project/Area Number: 15H02548
• Research Institution: Fukuoka University
• Principal Investigator: 廣瀬 伸一 福岡大学, 医学部, 教授 (60248515)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Keywords: 小児神経 / てんかん / イオンチャネル / iPS細胞 / 実験動物 / ラット

Outline of Annual Research Achievements

申請者らが設立・管理するてんかん遺伝子バンクと次世代シークエンサーを利用して変異をスクリーニングした。
その結果、主に乳児早期てんかん性の脳症で新規遺伝子を含め多数の遺伝子変異を発見した。さらにてんかん脳症の患者でパネルシークエンスにても遺伝子が同定出来なかった患者200名以上をトリオでエクソームシークエンスを終了し、新規遺伝子の遺伝子異常を同定している。　動物作出としては、KCNQ2脳症とKCNQ2良性てんかんの遺伝子変異が同じアミノ酸に起こる場合があるが、両方の遺伝子改変マウスを作出することに成功した。また既存のてんかんラットよりある遺伝子変異が関係している知見を得た。SCN1A遺伝子異常を持つDravet症候群の患者2名から新たにiPS細胞を樹立した。さらにすでに樹立していたDravet症候群の患者から樹立したiPS細胞を遺伝子編集技術をつかうことにより、“人工健常iPS細胞”を作出し報告した。さらにiPS細胞を特異的にグルタミン酸作動性の興奮性ニューロンまたはGABA作動性の抑制性ニューロンに分化誘導させる方法を用いて、Dravet症候群では興奮性ニューロンの電気活動は変わりないが、抑制性ニューロンの応答が悪いことを示した。これを利用して、多電極解析装置を使って薬物のrepositioningを実施中である。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Different types of suppression-burst patterns in patients with epilepsy of infancy with migrating focal seizures (EIMFS). (2019)
  • Author(s): Yoshitomi S, Takahashi Y, Imai K, Koshimizu E, Miyatake S, Nakashima M, Saitsu H, Matsumoto N, Kato M, Fujita T, Ishii A, Hirose S, Inoue Y.
  • Journal Title: Seizure. Volume: 65 Pages: 118-23
  • DOI: 10.1016/j.seizure.2019.01.009
  • Peer Reviewed
• [Journal Article] Generation of D1-1 TALEN isogenic control cell line from Dravet syndrome patient iPSCs using TALEN-mediated editing of the SCN1A gene. (2018)
  • Author(s): Tanaka Y, Sone T, Higurashi N, Sakuma T, Suzuki S, Ishikawa M, Yamamoto T, Mitsui J, Tsuji H, Okano H, Hirose S.
  • Journal Title: Stem Cell Res. Volume: 28 Pages: 100-4.
  • DOI: 10.1016/j.scr.2018.01.036
  • Peer Reviewed
• [Journal Article] Establishment of a human induced stem cell line (FUi002-A) from Dravet syndrome patient carrying heterozygous R1525X mutation in SCN1A gene. (2018)
  • Author(s): Tanaka Y, Higurashi N, Shirasu N, Yasunaga S, Moreira KM, Okano H, Hirose S.
  • Journal Title: Stem Cell Res. Volume: 31 Pages: 11-5.
  • DOI: 10.1016/j.scr.2018.06.008
  • Peer Reviewed
• [Journal Article] A patient with a GNAO1 mutation with decreased spontaneous movements, hypotonia, and dystonic features. (2018)
  • Author(s): Okumura A, Maruyama K, Shibata M, Kurahashi H, Ishii A, Numoto S, Hirose S, Kawai T, Iso M, Kataoka S, Okuno Y, Muramatsu H, Kojima S.
  • Journal Title: Brain Dev. Volume: 40(10) Pages: 926-30.
  • DOI: 10.1016/j.braindev.2018.06.005
  • Peer Reviewed
• [Journal Article] Somatic mosaic deletions involving SCN1A cause Dravet syndrome. (2018)
  • Author(s): Nakayama T, Ishii A, Yoshida T, Nasu H, Shimojima K, Yamamoto T, Kure S, Hirose S.
  • Journal Title: Am J Med Genet A. Volume: 176(3) Pages: 657-62.
  • DOI: 10.1002/ajmg.a.38596
  • Peer Reviewed
• [Journal Article] Long-term follow up of an adult with alternating hemiplegia of childhood and a p.Gly755Ser mutation in the ATP1A3 gene. (2018)
  • Author(s): Ito T, Narugami M, Egawa K, Yamamoto H, Asahina N, Kohsaka S, Ishii A, Hirose S, Shiraishi H.
  • Journal Title: Brain Dev. Volume: 40(3) Pages: 226-8
  • DOI: 10.1016/j.braindev.2017.11.007
  • Peer Reviewed
• [Journal Article] CACNA1A-related early-onset encephalopathy with myoclonic epilepsy: A case report. (2018)
  • Author(s): Hayashida T, Saito Y, Ishii A, Yamada H, Itakura A, Minato T, Fukuyama T, Maegaki Y, Hirose S.
  • Journal Title: Brain Dev. Volume: 40(2) Pages: 130-3.
  • DOI: 10.1016/j.braindev.2017.08.006
  • Peer Reviewed
• [Journal Article] Further characterization of CAPOS/CAOS syndrome with the Glu818Lys mutation in the ATP1A3 gene: A case report. (2018)
  • Author(s): Hayashida T, Saito Y, Ishii A, Hirose S, Hiraiwa R, Maegaki Y, Ohno K.
  • Journal Title: Brain Dev. Volume: 40(7) Pages: 576-81
  • DOI: 10.1016/j.braindev.2018.03.004
  • Peer Reviewed
• [Journal Article] Variant Intestinal-Cell Kinase in Juvenile Myoclonic Epilepsy. (2018)
  • Author(s): Bailey JN, de Nijs L, Bai D, Suzuki T, Miyamoto H, Tanaka M, Patterson C, Lin YC, Medina MT, Alonso ME, Serratosa JM, Duron RM, Nguyen VH, Wight JE, Martinez-Juarez IE, Ochoa A, Jara-Prado A, Guilhoto L, Molina Y, Yacubian EM, Lopez-Ruiz M, Inoue Y, Kaneko S, Hirose S, et al.
  • Journal Title: N Engl J Med Volume: 378(11) Pages: 1018-28
  • DOI: 10.1056/NEJMoa1700175
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Genetic background of developmental and epileptic encephalopathies (2018)
  • Author(s): Hirose S
  • Organizer: KEC2018 23rd
  • Int'l Joint Research
• [Presentation] Challenges and Opportunities in Pediatric Neurological Disease in Low Income Countries like Nepal (2018)
  • Author(s): Hirose S
  • Organizer: XIXth Nepalese Congress of Paediatrics,NEPCON 2018 & 37th Anniversary NEPAS
  • Int'l Joint Research / Invited
• [Presentation] Precision medicine through stem cells (2018)
  • Author(s): Hirose S,
  • Organizer: 12th Asian and Oceanian Epilepsy Congress
  • Int'l Joint Research
• [Presentation] Disease Modeling of Refractory Epilepsy using iPSCｓ (2018)
  • Author(s): Hirose S
  • Organizer: The 40th Annual Meeting of Japanese Society of Biological Psychiatry The 61st Annual Meeting of the Japanese Society for Neurochmistry
  • Int'l Joint Research
• [Presentation] Genes in Focal Epilepsy (2018)
  • Author(s): Hirose S
  • Organizer: ANZCNS 7th Annual Scientific Meeting
  • Int'l Joint Research
• [Presentation] Application of Genetics to Epilepsy after Gene-Hunting (2018)
  • Author(s): Hirose S
  • Organizer: Anzcns 7th Annual Scientific Meeting
  • Int'l Joint Research
• [Presentation] Pathogenesis and genetic substrate (2018)
  • Author(s): Hirose S
  • Organizer: 19th International symposium on severe infantile epilepsies: old and new treatments.
  • Int'l Joint Research