Investigation of cellular/molecular pathophysiology of brain disorders using human brain organoids established from iPS cells

• Project/Area Number: 16K06996
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Kumamoto University
• Principal Investigator: Numakawa Tadahiro 熊本大学, 発生医学研究所, 特定事業研究員 (40425690)
• Research Collaborator: ODAKA HARUKI
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2018: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: iPS細胞 / 大脳皮質 / 脳疾患 / 神経細胞 / オルガノイド / iPS / 神経科学 / 脳神経疾患

Outline of Final Research Achievements

Although studies using neurons obtained from animals including rats or mice are very useful to examine the cellular/molecular basis of brain disorders, models using human neurons established from iPSCs are much more suitable for investigating the pathophysiology of brain disease. In this study, dissociated human neurons and brain organoids made using iPSCs (healthy, maple syrup urine disease, propionic academia, and adrenoleukodystrophy) were established. Interestingly, disease human neurons (from iPSC of maple syrup urine disease or propionic academia) showed a significant decrease in the system of neurotransmitter release. Furthermore, it was revealed that brain organoids of maple syrup urine disease exhibited an immaturity in the neuronal differentiation.

Academic Significance and Societal Importance of the Research Achievements

これまでの脳研究では、生きたヒト脳を使用することは極めて困難であったため、動物個体レベルもしくは動物脳より分散させた初代培養ニューロンを用いたアプローチが主であった。本研究では、これまでの動物を用いた研究の弱点を克服するiPS細胞由来のヒトニューロンを用いて、メープルシロップ尿症、副腎白質ジストロフィー、およびプロピオン酸血症などの有効な治療法がみつかっていない疾患において、細胞・分子レベルでの病態解析に成功した。特に、メープルシロップ尿症やプロピオン酸血症では、機能的ニューロンにおいて、それぞれ固有の脆弱性を発見し、これが今後の病態解明および新規薬剤スクリーニングに役立つ可能性を示した。

Research Products

• [Journal Article] Basic fibroblast growth factor increased glucocorticoid receptors in cortical neurons through MAP kinase pathway (2018)
  • Author(s): Numakawa Tadahiro、Odaka Haruki、Adachi Naoki、Chiba Shuichi、Ooshima Yoshiko、Matsuno Hitomi、Nakajima Shingo、Yoshimura Aya、Fumimoto Kazuhiro、Hirai Yohei、Kunugi Hiroshi
  • Journal Title: Neurochemistry International Volume: 118 Pages: 217-224
  • DOI: 10.1016/j.neuint.2018.06.009
  • Peer Reviewed
• [Journal Article] Actions of Brain-Derived Neurotrophin Factor in the Neurogenesis and Neuronal Function, and Its Involvement in the Pathophysiology of Brain Diseases (2018)
  • Author(s): Numakawa Tadahiro、Odaka Haruki、Adachi Naoki
  • Journal Title: International Journal of Molecular Sciences Volume: 19 Issue: 11 Pages: 3650-3650
  • DOI: 10.3390/ijms19113650
  • Peer Reviewed / Open Access
• [Journal Article] The Sox2 promoter-driven CD63-GFP transgenic rat model allows tracking of neural stem cell-derived extracellular vesicles. (2018)
  • Author(s): Yoshimura A., Adachi N., Matsuno H., Kawamata M., Yoshioka Y., Kikuchi H., Odaka H., Numakawa T., Kunugi H., Ochiya T.* & Tamai Y.
  • Journal Title: Disease Models & Mechanisms Volume: 11 Issue: 1 Pages: 028779-028779
  • DOI: 10.1242/dmm.028779
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Actions of Brain-Derived Neurotrophic Factor and Glucocorticoid Stress in Neurogenesis. (2017)
  • Author(s): Numakawa T.*, Odaka H. & Adachi N.
  • Journal Title: International Journal of Molecular Sciences Volume: 18 Issue: 11 Pages: 2312-2312
  • DOI: 10.3390/ijms18112312
  • Peer Reviewed / Open Access
• [Journal Article] Impact of glucocorticoid on neurogenesis. (2017)
  • Author(s): Odaka H., Adachi N. & Numakawa T.
  • Journal Title: Neural Regeneration Research Volume: 12 Issue: 7 Pages: 1028-1035
  • DOI: 10.4103/1673-5374.211174
  • Peer Reviewed / Open Access
• [Journal Article] Generation of a novel transgenic rat model for tracing extracellular vesicles in body fluids. (2016)
  • Author(s): Yoshimura A, Kawamata M, Yoshioka Y, Katsuda T, Kikuchi H, Nagai Y, Adachi N, Numakawa T, Kunugi H, Ochiya T, Tamai Y.
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 31172-31172
  • DOI: 10.1038/srep31172
  • Peer Reviewed / Open Access
• [Journal Article] Chronic glucocorticoid exposure suppressed the differentiation and survival of embryonic neural stem/progenitor cells: Possible involvement of ERK and PI3K/Akt signaling in the neuronal differentiation. (2016)
  • Author(s): Odaka H, Numakawa T, Yoshimura A, Nakajima S, Adachi N, Ooshima Y, Inoue T, Kunugi H.
  • Journal Title: Neuroscience Research Volume: 113 Pages: 28-36
  • DOI: 10.1016/j.neures.2016.07.002
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Negative regulation of microRNA-132 in expression of synaptic proteins in neuronal differentiation of embryonic neural stem cells. (2016)
  • Author(s): Aya Yoshimura, Tadahiro Numakawa, Haruki Odaka, Naoki Adachi, Yoshitaka Tamai, Hiroshi Kunugi
  • Journal Title: Neurochemistry International Volume: なし Pages: 26-33
  • DOI: 10.1016/j.neuint.2016.04.013
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] SIALIDOSIS NEURONS FROM PATIENT-DERIVED INDUCED PLURIPOTENT STEM CELL EXHIBIT SYNAPTIC DYSFUNCTION (2018)
  • Author(s): Haruki Odaka, Tadahiro Numakawa, Minami Soga, Jun Kido, Ryutaro Kajihara, Toshika Okumiya, Hirokazu Furuya, Takafumi Inoue, Takumi Era
  • Organizer: ISSCR 2018 - The International Society of Stem Cell Research Annual Meeting
  • Int'l Joint Research
• [Presentation] ゴーシェー病II 型患者由来のiPS 細胞を使用した薬剤スクリーニングシステムの開発 (2017)
  • Author(s): 城戸 淳、沼川 忠広、松本 志郎、小高 陽樹、曽我 美南、梶原 隆太郎、 遠藤 文夫、中村 公俊、江良 択実
  • Organizer: 第59回日本先天性代謝異常学会総会
• [Presentation] シアリドーシス患者iPS 細胞を用いた神経障害の病理学的解析 (2017)
  • Author(s): 小髙 陽樹、沼川 忠広、曽我 美南、城戸 淳、梶原 隆太郎、奥宮 敏可、古谷博和、井上 貴文、江良 択実
  • Organizer: 第59回日本先天性代謝異常学会総会