新規遺伝子改変動物と患者ｉＰＳを利用した分子病態に基づくてんかんの革新的治療開発

2014 Fiscal Year Annual Research Report

• Project/Area Number: 24249060
• Research Institution: Fukuoka University
• Principal Investigator: 廣瀬 伸一 福岡大学, 医学部, 教授 (60248515)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: てんかん / ドラベ症候群 / 遺伝子 / 遺伝子改変動物 / 薬剤スクリーニング

Outline of Annual Research Achievements

本研究は遺伝学的背景が要因と考えられる“てんかん症候群”と“けいれん疾患”の責任遺伝子を同定し発症機序の解明と利用戦略基盤を構築することを目的とした。てんかんの病態は長い間不明であったが、最近の分子生物学の発展に伴い遺伝子異常が発見されるようになった。しかしながら、その遺伝子異常がヒト脳内でどのような病態を引き起こしているのかは、ヒト脳を利用する他なく困難と考えられてきた。ところが、最近は技術革新により、ヒト脳での病態研究が望める可能性が生まれた。
我々はすでに500種以上のてんかんの責任遺伝子変異を見出してきた。また次世代シークエンサ－を利用して、世界に先駆けてんかんを来す小児交互性片麻痺の責任遺伝子を明らかにした。これら変異による病態を電気生理学的、細胞生物学的に明らかにしてきた。特筆すべきはナトリリウムチャネルをコードする遺伝子、ＳＣＮ１Ａの異常による起こるドラベ症候群の患者より樹立したiPS細胞から、神経を分化させすることに世界に先駆け成功した。これにより、ドラベ症候群の分子病態を明らかにすることができた。さらにこの細胞を用いて、そのＳＣＮ１Ａの異常をＴＡＬＥＮによる方法で、修復また正常細胞に導入することに成功した。
てんかんでの遺伝子異常の発見により、その遺伝子異常をもつ動物を、遺伝子操作で作出することができるようになった。我々はドラベ症候群でみられるようなＳＣＮ1Ａの遺伝子の微少欠失を持つマウスの作出に成功した。さらにＴＡＬＥＮ技術を使い、ラットのＳＣＮ1Ａの遺伝子とＰＣＤＨ19に変異を導入することに成功した。
現在、樹立したiPS細胞を用いて、病態の回復を指標として多種の薬剤をスクリーニングすることにより、効果のある薬剤のシーズの探索を行っている。同様に作出した動物を用いて、病態に基づく治療薬のシーズとなる薬剤のスクリーニングを実施している。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Single Nucleotide Variations in CLCN6 Identified in Patients with Benign Partial Epilepsies in Infancy and/or Febrile Seizures. (2015)
  • Author(s): Yamamoto T, Shimojima K, Sangu N, Komoike Y, Ishii A, Abe S, Yamashita S, Imai K, Kubota T, Fukasawa T, Okanishi T, Enoki H, Tanabe T, Saito A, Furukawa T, Shimizu T, Milligan CJ, Petrou S, Heron SE, Dibbens LM, Hirose S, Okumura A.
  • Journal Title: PLoS ONE Volume: 10(3) Pages: e0118946
  • DOI: 10.1371/journal.pone.0118946
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A case of recurrent encephalopathy with SCN2A missense mutation. (2015)
  • Author(s): Fukasawa T, Kubota T, Negoro T, Saitoh M, Mizuguchi M, Ihara Y, Ishii A, Hirose S.
  • Journal Title: Brain Dev Volume: 37(6) Pages: 631-4
  • DOI: 10.1016/j.braindev.2014.10.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] JNK is critical for the development of Candida albicans-induced vascular lesions in a mouse model of Kawasaki disease. (2015)
  • Author(s): Yoshikane Y, Koga M, Imanaka-Yoshida K, Cho T, Yamamoto Y, Yoshida T, Hashimoto J, Hirose S, Yoshimura K.
  • Journal Title: Cardiovasc Pathol. Volume: 24(1) Pages: 33-40
  • DOI: 10.1016/j.carpath.2014.08.005
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Trans-Golgi protein p230/golgin-245 is involved in phagophore formation. (2015)
  • Author(s): Sohda M, Misumi Y, Ogata S, Sakisaka S, Hirose S, Ikehara Y, Oda K.
  • Journal Title: Biochem Biophys Res Commun. Volume: 456(1) Pages: 275-81
  • DOI: 10.1016/j.bbrc.2014.11.071
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Clinical and genetic features of acute encephalopathy in children taking theophylline. (2015)
  • Author(s): Saitoh M, Shinohara M, Ishii A, Ihara Y, Hirose S, Shiomi M, Kawawaki H, Kubota M, Yamagata T, Miyamoto A, Yamanaka G, Amemiya K, Kikuchi K, Kamei A, Akasaka M, Anzai Y, Mizuguchi M.
  • Journal Title: Brain Dev Volume: 37 Pages: 463-70
  • DOI: 10.1016/j.braindev.2014.07.010
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Effect of CYP2C19 polymorphisms on stiripentol administration in Japanese cases of Dravet syndrome. (2015)
  • Author(s): Kouga T, Shimbo H, Iai M, Yamashita S, Ishii A, Ihara Y, Hirose S, Yamakawa K, Osaka H.
  • Journal Title: Brain Dev Volume: 37(2) Pages: 243-9
  • DOI: 10.1016/j.braindev.2014.04.003
  • Peer Reviewed
• [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] The kick-in system: a novel rapid knock-in strategy. (2014)
  • Author(s): Tomonoh Y, Deshimaru M, Araki K, Miyazaki Y, Arasaki T, Tanaka Y, Kitamura H, Mori F, Wakabayashi K, Yamashita S, Saito R, Itoh M, Uchida T, Yamada J, Migita K, Ueno S, Kitaura H, Kakita A, Lossin C, Takano Y, Hirose S.
  • Journal Title: PLoS ONE Volume: 9(2) Pages: e88549
  • DOI: :10.1371/journal.pone.0088549
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Elfn1 recruits presynaptic mGluR7 in trans and its loss results in seizures. (2014)
  • Author(s): Tomioka NH, Yasuda H, Miyamoto H, Hatayama M, Morimura N, Matsumoto Y, Suzuki T, Odagawa M, Odaka YS, Iwayama Y, Won Um J, Ko J, Inoue Y, Kaneko S, Hirose S, Yamada K, Yoshikawa T, Yamakawa K, Aruga J.
  • Journal Title: Nat Commun Volume: 5 Pages: 4501
  • DOI: 10.1038/ncomms5501
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Genotype-phenotype correlations in alternating hemiplegia of childhood. (2014)
  • Author(s): Sasaki M, Ishii A, Saito Y, Morisada N, Iijima K, Takada S, Araki A, Tanabe Y, Arai H, Yamashita S, Ohashi T, Oda Y, et al 34番目
  • Journal Title: Neurology Volume: 82(6) Pages: 482-90.
  • DOI: 10.1212/WNL.0000000000000102
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Intermediate form between alternating hemiplegia of childhood and rapid-onset dystonia-parkinsonism. (2014)
  • Author(s): Sasaki M, Ishii A, Saito Y, Hirose S.
  • Journal Title: Mov Disord. Volume: 29(1) Pages: 153-4
  • DOI: 10.1002/mds.25659
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Exacerbation of benign familial neonatal epilepsy induced by massive doses of phenobarbital and midazolam. (2014)
  • Author(s): Maeda T, Shimizu M, Sekiguchi K, Ishii A, Ihara Y, Hirose S, Izumi T.
  • Journal Title: Pediatr Neurol Volume: 51(2) Pages: 259-61
  • DOI: 10.1016/j.pediatrneurol.2014.04.004
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Obesity attenuates D2 autoreceptor-mediated inhibition of putative ventral tegmental area dopaminergic neurons. (2014)
  • Author(s): Koyama S, Mori M, Kanamaru S, Sazawa T, Miyazaki A, Terai H, Hirose S.
  • Journal Title: Physiol Rep Volume: 2(5) Pages: e12004
  • DOI: 10.14814/phy2.12004
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A novel SCN1A mutation in a cytoplasmic loop in intractable juvenile myoclonic epilepsy without febrile seizures. (2014)
  • Author(s): Jingami N, Matsumoto R, Ito H, Ishii A, Ihara Y, Hirose S, Ikeda A, Takahashi R.
  • Journal Title: Epileptic Disord Volume: 16(2) Pages: 227-31
  • DOI: 10.1684/epd.2014.0657
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Association of nonsense mutation in GABRG2 with abnormal trafficking of GABAA receptors in severe epilepsy. (2014)
  • Author(s): Ishii A, Kanaumi T, Sohda M, Misumi Y, Zhang B, Kakinuma N, Haga Y, Watanabe K, Takeda S, Okada M, Ueno S, Kaneko S, Takashima S, Hirose S.
  • Journal Title: Epilepsy Res. Volume: 108(3) Pages: 420-32
  • DOI: 10.1016/j.eplepsyres.2013.12.005
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Early onset and focal spike discharges as indicators of poor prognosis for myoclonic-astatic epilepsy. (2014)
  • Author(s): Inoue T, Ihara Y, Tomonoh Y, Nakamura N, Ninomiya S, Fujita T, Ideguchi H, Yasumoto S, Zhang B, Hirose S.
  • Journal Title: Brain Dev Volume: 36(7) Pages: 613-9.
  • DOI: 10.1016/j.braindev.2013.08.009
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Case of Desbuquois dysplasia type 1: potentially lethal skeletal dysplasia. (2014)
  • Author(s): Inoue S, Ishii A, Shirotani G, Tsutsumi M, Ohta E, Nakamura M, Mori T, Inoue T, Nishimura G, Ogawa A, Hirose S.
  • Journal Title: Pediatr Int. Volume: 56(4) Pages: e26-9
  • DOI: 10.1111/ped.12383
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Distinct neurological disorders with ATP1A3 mutations. (2014)
  • Author(s): Heinzen EL, Arzimanoglou A, Brashear A, Clapcote SJ, Gurrieri F, Goldstein DB, Johannesson SH, Mikati MA, Neville B, Nicole S, Ozelius LJ, Poulsen H, Schyns T, Sweadner KJ, van den Maagdenberg A, Vilsen B, for the ATPAWG, (Hirose S, et al, ).
  • Journal Title: Lancet Neurol Volume: 13(5) Pages: 503-14
  • DOI: 10.1016/S1474-4422(14)70011-0
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] てんかんとiPS細胞 (2014)
  • Author(s): 廣瀨伸一
  • Organizer: 第44回日本臨床神経生理学会
  • Place of Presentation: 福岡
  • Year and Date: 2014-11-19 – 2014-11-21
• [Presentation] A recurrent KCNT1 mutation in two sporadic cases with malignant migrating partial seizures in infancy (2014)
  • Author(s): Okumura A, Ishii A, Shioda M, Kidokoro H, Sakauchi, M, Shimada S, Shimizu T, Osawa M, Hirose S, Yamamaoto T.
  • Organizer: The 16th annual meeting of the infantile seizure society
  • Place of Presentation: TURKEY
  • Year and Date: 2014-06-22 – 2014-06-27
• [Presentation] A neonatal case of malignant migrating partial seizures in infancy (2014)
  • Author(s): Sato T, Watanabe Y, Watanabe K, Yamashita M, Hashimoto K, Dateki S, Shirakawa T, Nakashima Y, Ihara Y, Ishii A, Hirose S, Moriuchi H
  • Organizer: The 16th annual meeting of the infantile seizure society
  • Place of Presentation: TURKEY
  • Year and Date: 2014-06-22 – 2014-06-27
• [Presentation] THE pathomechanisms of Dravet syndome:Lessons From patient-Derived Induced Pluripotent steｍ Cells(iPSCｓ） (2014)
  • Author(s): Hirose S
  • Organizer: The 16th annual meeting of the infantile seizure society
  • Place of Presentation: TURKEY
  • Year and Date: 2014-06-22 – 2014-06-27
  • Invited
• [Presentation] 最新技術でてんかんへ挑む (2014)
  • Author(s): 廣瀨伸一
  • Organizer: 第217回日本小児科学会宮城地方会
  • Place of Presentation: 宮城
  • Year and Date: 2014-06-08
• [Presentation] Newborn screening for Fabry disease:Prevalence and genotypes from a pilot study in Japan (2014)
  • Author(s): Inoue T, Hattori K, Ihara K, Ishii A, Nakamura K, Hirose S.
  • Organizer: The 2nd Asian Congress for Lysosomal Storage
  • Place of Presentation: 福岡
  • Year and Date: 2014-06-06
• [Book] Genetics of Epilepsy (2014)
  • Author(s): Hirose S
  • Total Pages: 31
  • Publisher: Prog Brain Res.
• [Book] 小児科研修ノート 第2版 (2014)
  • Author(s): 廣瀨伸一
  • Total Pages: 3
  • Publisher: 診断と治療社