Research on Dravet syndrome by using patient-derived iPS cells and a rat model

• Project/Area Number: 26461552
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Jikei University School of Medicine
• Principal Investigator: HIGURASHI NORIMICHI 東京慈恵会医科大学, 医学部, 講師 (40568820)
• Research Collaborator: TAHARA Mayu 東京慈恵会医科大学, 医学部, 大学院生
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: てんかん / 病態 / 細胞治療 / iPS細胞 / モデルラット / MRI / 遺伝子 / 創薬

Outline of Final Research Achievements

Dravet syndrome (DS) is an infantile-onset intractable epilepsy, mainly caused by mutation in SCN1A gene. This research aims to elucidate novel pathomechanisms on DS and to achieve a transplantation therapy of GABAergic inhibitory precursors in the future. In the present study, 1) we developed a method to efficiently induce GABAergic neurons from human induced pluripotent stem cells (iPSCs). 2) We established mutation-repaired iPSCs from a patient-derived iPSCs by using a TALEN-based genome-editing technology. 3) Finally, to identify brain regions that is involved in the epilepsy pathogenesis in DS, we employed a manganese-enhanced magnetic resonance imaging technique, which may detect regions with increased neuronal activity. We identified several enhanced regions including hippocampus, thalamus, and some neocortical areas. Although further evaluations are necessary, this method will elucidate anatomical basis for epileptogenesis and target regions for cell therapy.

Research Products

• [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 Yasuyoshi、Sone Takefumi、Higurashi Norimichi、Sakuma Tetsushi、Suzuki Sadafumi、Ishikawa Mitsuru、Yamamoto Takashi、Mitsui Jun、Tsuji Hitomi、Okano Hideyuki、Hirose Shinichi
  • Journal Title: Stem Cell Research Volume: 28 Pages: 100-104
  • DOI: 10.1016/j.scr.2018.01.036
  • Peer Reviewed / Open Access
• [Journal Article] Characteristic phasic evolution of convulsive seizure in PCDH19-related epilepsy. (2016)
  • Author(s): 1.Ikeda H, Imai K, Ikeda H, Shigematsu H, Takahashi Y, Inoue Y, Higurashi N, Hirose S
  • Journal Title: Epileptic Disord Volume: 18 Issue: 1 Pages: 26-33
  • DOI: 10.1684/epd.2016.0803
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A patient with an early diagnosis of PCDH19-related epilepsy (2015)
  • Author(s): 保科めぐみ，日暮憲道，阿部優作，他
  • Journal Title: NO TO HATTATSU Volume: 47 Issue: 4 Pages: 305-309
  • DOI: 10.11251/ojjscn.47.305
  • NAID: 130005111608
  • ISSN: 0029-0831, 1884-7668
  • Peer Reviewed
• [Journal Article] Efficacy of antiepileptic drugs for the treatment of Dravet syndrome with different genotypes. (2015)
  • Author(s): Shi XY, Tomonoh Y, Wang WZ, Ishii A, Higurashi N, et al.
  • Journal Title: Brain Dev Volume: 38 Issue: 1 Pages: 40-46
  • DOI: 10.1016/j.braindev.2015.06.008
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Characterizing PCDH19 in human induced pluripotent stem cells (iPSCs) and iPSC-derived developing neurons: emerging role of a protein involved in controlling polarity during neurogenesis. (2015)
  • Author(s): Compagnucci C, Petrini S, Higuraschi N, et al.
  • Journal Title: Oncotarget Volume: 6 Issue: 29 Pages: 26804-26813
  • DOI: 10.18632/oncotarget.5757
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Immediate suppression of seizure clusters by corticosteroids in PCDH19 female epilepsy. (2015)
  • Author(s): Higurashi N, Takahashi Y, Kashimada A, Sugawara Y, Sakuma H, Tomonoh Y, Inoue T, Hoshina M, Satomi R, Ohfu M, Itomi K, Takano K, Kirino T, Hirose S.
  • Journal Title: Seizure Volume: 27 Pages: 1-5
  • DOI: 10.1016/j.seizure.2015.02.006
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Clinical utility of neuronal cells directly converted from fibroblasts of patients for neuropsychiatric disorders: studies of lysosomal storage diseases and channelopathy. (2015)
  • Author(s): Kano S, Yuan M, Cardarelli RA, Maegawa G, Higurashi N, et al.
  • Journal Title: Currrent Molecular Medicine Volume: 15 Issue: 2 Pages: 138-145
  • DOI: 10.2174/1566524015666150303110300
  • Peer Reviewed
• [Presentation] Overview of iPSC research on pediatric neurological diseases. (2017)
  • Author(s): Higurashi N
  • Organizer: 14th Asian and Oceanian Congress of Child Neurology
  • Int'l Joint Research / Invited
• [Presentation] 前頭葉てんかんが疑われた2例における睡眠時Minor motor eventの臨床的意義 (2017)
  • Author(s): 日暮憲道、菊池健二郎
  • Organizer: 第59回日本小児神経学会総会
• [Presentation] 患者神経細胞を用いたDravet症候群の病態・治療研究 (2017)
  • Author(s): 日暮憲道
  • Organizer: 第51回日本てんかん学会学術総会
  • Invited
• [Presentation] Generation of heterozygous (Pcdh19+/－) female rats as a model for PCDH19-related epilepsy (2016)
  • Author(s): Higurashi N, Takata F, Goto A, Uchida T, Ohmori I, Mashimo T, Kataoka Y, Hirose S
  • Organizer: 第50回日本てんかん学会学術総会
  • Place of Presentation: 静岡
  • Year and Date: 2016-10-08
• [Presentation] PCDH19関連てんかんの最新事情 (2016)
  • Author(s): 日暮憲道、廣瀬伸一
  • Organizer: 第58回日本小児神経学会総会
  • Place of Presentation: 東京
  • Year and Date: 2016-06-04
  • Invited
• [Presentation] PCDH19 Female Epilepsy (2016)
  • Author(s): 1.Higurashi N, Hirose S
  • Organizer: 53rd Annual Conference of Indian Academy of Pediatrics (PEDICON), 15th Asia Pacific Congress of Pediatrics (APCP)
  • Place of Presentation: Hyderabad, India
  • Year and Date: 2016-01-23
  • Int'l Joint Research / Invited
• [Presentation] 左片側の持続性部分てんかんを主徴とし、病初期にラスムッセン脳炎が疑われたがびまん性脳萎縮を呈した慢性脳炎の男児例 (2015)
  • Author(s): 日暮憲道、折津友隆、高橋幸利、佐久間啓、浜野晋一郎、井田博幸
  • Organizer: 第57回日本小児神経学会総会
  • Place of Presentation: 大阪
  • Year and Date: 2015-05-28
• [Presentation] Immediate suppression of seizure clusters by corticosteroids in PCDH19 female epilepsy. (2015)
  • Author(s): Higurashi N, Takahashi Y, Hirose S
  • Organizer: 13th Asian and Oceanian Congress of Child Neurology (AOCCN 2015)
  • Place of Presentation: Taipei, Taiwan
  • Year and Date: 2015-05-16
  • Int'l Joint Research
• [Presentation] New technologies in epilepsy research (2015)
  • Author(s): Higurashi N
  • Organizer: The 11th Congress of Asian Society for Pediatric Research (ASPR 2015)
  • Place of Presentation: Osaka
  • Year and Date: 2015-04-16
  • Int'l Joint Research / Invited
• [Presentation] Dravet syndrome research using patient-derived induced pluripotent stem cells—Present findings and future directions. (2014)
  • Author(s): Higurashi N, Hirose S
  • Organizer: Korea Epilepsy Congress (KEC 2014)
  • Place of Presentation: ソウル（韓国）
  • Year and Date: 2014-06-14
  • Invited
• [Presentation] シンポジウム iPS細胞を用いた小児神経疾患の病態解明への期待とその問題点 ②てんかんの病態研究とその問題点 (2014)
  • Author(s): 日暮憲道
  • Organizer: 第56回日本小児神経学会総会
  • Place of Presentation: 浜松
  • Year and Date: 2014-05-29
  • Invited
• [Book] 稀少てんかんの診療指標 (2017)
  • Author(s): 日暮憲道
  • Total Pages: 276
  • Publisher: 診断と治療社
• [Book] 新薬と臨床 (2017)
  • Author(s): 日暮憲道, 廣瀬伸一
  • Total Pages: 100
  • Publisher: 医薬情報研究所
• [Book] 小児科診療増刊号 小児の症候群 (2016)
  • Author(s): 日暮憲道
  • Total Pages: 416
  • Publisher: 診断と治療社
• [Book] 医学のあゆみ てんかんの遺伝子診断 (2015)
  • Author(s): 日暮憲道
  • Total Pages: 70
  • Publisher: 医歯薬出版
• [Book] 遺伝子医学MOOK 27号 iPS細胞を用いた難病研究－臨床病態解明と創薬に向けた研究の最新知見 (2015)
  • Author(s): 日暮憲道, 廣瀬伸一
  • Total Pages: 228
  • Publisher: メディカル ドゥ
• [Book] 症例から学ぶ 戦略的てんかん診断・治療 (2014)
  • Author(s): 日暮憲道，井原由紀子，廣瀬伸一
  • Total Pages: 242
  • Publisher: 南山堂
• [Book] 臨床てんかん学
  • Author(s): 日暮憲道, 廣瀬伸一
  • Total Pages: 688
  • Publisher: 医学書院