In vitro brain development analysis using iPS cells and genome editing technologies

• Project/Area Number: 26830018
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Keio University
• Principal Investigator: Ishikawa Mitsuru 慶應義塾大学, 医学部(信濃町), 特任助教 (10613995)
• Research Collaborator: TANAKA Shoma
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
• Keywords: iPS 細胞 / ゲノム編集 / 大脳皮質 / オルガノイド / 神経発達 / 進化 / iPS細胞 / 霊長類 / 分節重複 / 重複遺伝子 / 興奮性神経細胞 / 進化生物学 / SRGAP2 / 細胞移動 / Neurogenin2 / 不死化リンパ芽球

Outline of Final Research Achievements

Toward estimating characteristics of human or non-human primate-specific genes or their specific CNVs, we are investigating how the annotated genes function in neuronal development using human induced pluripotent stem cells and genome editing techniques. In this study, we have developed two efficient neural induction methods from human iPS cells. One is purification of excitatory neurons in a dish, by which biochemical analysis became to well function because of its less heterogeneity. On the other hand, generating human cerebral organoid system have been developed. Using CRSPR/Cas9 to human genome, we demonstrated that knock out of a neural development and microcephaly-related gene generated abnormal construction of human cortical layers. In summary, our studies would become a support of identifying and evaluating therapeutic strategies to ameliorate the symptoms of neurodevelopmental pathogenesis.

Research Products

• [Journal Article] Establishment of in vitro FUS-associated familial amyotrophic lateral sclerosis model using human induced pluripotent stem cells. (2016)
  • Author(s): Naoki Ichiyanagi, Koki Fujimori, Masato Yano, Chikako Ishihara-Fujisaki, Takefumi Sone, Tetsuya Akiyama, Yohei Okada, Wado Akamatsu, Takuya Matsumoto, Mitsuru Ishikawa, Yoshinori Nishimoto, Yasuharu Ishihara, Tetsushi Sakuma, Takashi Yamamoto, Hitomi Tsuiji, Naoki Suzuki, Hitoshi Warita, Masashi Aoki, Hideyuki Okano
  • Journal Title: Stem Cell Reports Volume: 6 Issue: 4 Pages: 496-510
  • DOI: 10.1016/j.stemcr.2016.02.011
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functional Neurons Generated from T Cell-Derived Induced Pluripotent Stem Cells (2016)
  • Author(s): Matsumoto T, Fujimori K, Andoh-Noda T, Ando T, Kuzumaki N, Toyoshima M, Tada H, Imaizumi K, Ishikawa M, Yamaguchi R, Isoda M, Zhou Z, Sato S, Kobayashi T, Ohtaka M, Nishimura K, Kurosawa H, Yoshikawa T, Takahashi T, Nakanishi M, Ohyama M, Hattori N, Akamatsu W, Okano H.
  • Journal Title: Stem Cell Reports. Volume: 6 Issue: 3 Pages: 422-35
  • DOI: 10.1016/j.stemcr.2016.01.010
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cellular localization and dendritic function of rat isoforms of the SRF coactivator MKL1 in cortical neurons. (2014)
  • Author(s): Ishikawa M, Shiota J, Ishibashi Y, Hakamata T, Shoji S, Fukuchi M, Tsuda M, Shirao T, Sekino Y, Baraban JM, Tabuchi A.
  • Journal Title: Neuroreport. Volume: 25 Issue: 8 Pages: 585-92
  • DOI: 10.1097/wnr.0000000000000141
  • Peer Reviewed
• [Presentation] Widespread analysis of TSSs in human iPSC-derived neurons using CAGE-SEQ (2016)
  • Author(s): 石川充
  • Organizer: 日本神経科学大会
  • Place of Presentation: パシフィコ横浜、神奈川
• [Presentation] Molecular and functional characterization of Angelman syndrome patient-derived iPSCs and neurons (2015)
  • Author(s): Mitsuru Ishikawa, Hironobu Okuno, Shoma Tanaka, Yuki Nakatake, Wado Akamatsu, Shinji Saitoh, Kenjiro Kosaki, Minoru Ko, Hideyuki Okano
  • Organizer: 29th International Mammalian Genome Conference
  • Place of Presentation: 横浜開港記念館（神奈川県横浜市）
  • Year and Date: 2015-11-08
  • Int'l Joint Research
• [Presentation] ヒトiPS細胞由来興奮性ニューロン分化誘導技術を用いた神経発達障害における遺伝子発現と細胞形態解析 (2015)
  • Author(s): 石川充、奥野博庸、田中翔真、中武悠樹、赤松和土、小崎健次郎、齋藤伸治、洪実、岡野栄之
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸コンベンションセンター （兵庫県神戸市）
  • Year and Date: 2015-07-28
• [Presentation] Expression analysis of imprinted genes in Angelman syndrome patient derived-iPSCs and iPSCs-derived neurons (2015)
  • Author(s): Mitsuru Ishikawa, Hironobu Okuno, Shoma Tanaka, Wado Akamatsu, Shinji Saitoh, Kenjiro Kosaki, Hideyuki Okano
  • Organizer: The 11th International Workshop on Advanced Genomics
  • Place of Presentation: 一ツ橋ホール（東京都千代田区）
  • Year and Date: 2015-05-20
  • Int'l Joint Research
• [Presentation] ヒトiPS細胞由来神経細胞における活動依存的遺伝子発現機構の解析 (2015)
  • Author(s): 石川充、赤松和土、駒野肇、岡野栄之
  • Organizer: 第１４回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-20
• [Presentation] 神経フェリチン症（Neuroferritinopathy）患者由来iPS細胞の樹立と病態モデルの検討 (2015)
  • Author(s): 森本悟、石川充、西田勝也、岡本理志、駒野肇、赤松和土、Holly J Garringer、Vidal Ruben、白石泰三、高尾昌樹、岡野栄之
  • Organizer: 第１４回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-20