Direct differentiation of peripheral blood cells into neurons for the analysis of epigenetic-related diseases

• Project/Area Number: 17K10083
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• 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)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: ダイレクトリプログラミング / iN / エピゲノム / 神経疾患 / 末梢血細胞 / 直接誘導 / iPS細胞 / 神経科学 / 脳神経疾患 / 発生・分化 / 脳・神経

Outline of Final Research Achievements

In this study, we have developed a technique for an efficient conversion of peripheral blood mononuclear cells into neurons using Sendai virus system. It is faster and more survival efficient than previously reported. This method produced glutamatergic neurons as the main population as the previous methods, but it also included GABAergic and dopaminergic neurons. These results suggested that it will be possible in the future to create selective neural subtypes by adjusting or adding another differentiation factors. In addition, we were successfully able to construct efficient platforms using high-content cell-image analyzers and a single-cell RNA sequencing for validating the character of the cells produced by the culture systems like this study.

Academic Significance and Societal Importance of the Research Achievements

本研究を通して末梢血細胞からiPS細胞を介することのない、極めて迅速な神経分化転換技術（ダイレクトリプログラミング）のプラットフォームができ、その詳細な性質も把握できるようになった。さらに、iPS細胞からの分化誘導時と導入している因子がほぼ同じであるにもかかわらず血球細胞からの分化転換ではより多岐のサブタイプの神経細胞が産生されている。これは細胞個々がもつエピジェネティック情報を完全にリプログラミングしていないことも大きな理由の一つであると予想される。そのため、これまで困難であったエピジェネティックに関与する疾患の病態評価や治療法開発に向けて、新しく機能的実験系を提唱できたと考えられる。

Research Products

• [Journal Article] miRNA-Based Rapid Differentiation of Purified Neurons from hPSCs Advancestowards Quick Screening for Neuronal Disease Phenotypes In Vitro (2020)
  • Author(s): Ishikawa Mitsuru、Aoyama Takeshi、Shibata Shoichiro、Sone Takefumi、Miyoshi Hiroyuki、Watanabe Hirotaka、Nakamura Mari、Morota Saori、Uchino Hiroyuki、Yoo Andrew S.、Okano Hideyuki
  • Journal Title: Cells Volume: 9 Issue: 3 Pages: 532-532
  • DOI: 10.3390/cells9030532
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] In Vitro Modeling of the Bipolar Disorder and Schizophrenia Using Patient-Derived Induced Pluripotent Stem Cells with Copy Number Variations of PCDH15 and RELN (2019)
  • Author(s): Ishii Takaya、Ishikawa Mitsuru、Fujimori Koki、Maeda Takuji、Kushima Itaru、Arioka Yuko、Mori Daisuke、Nakatake Yuhki、Yamagata Bun、Nio Shintaro、Kato Takahiro A.、Yang Nan、Wernig Marius、Kanba Shigenobu、Mimura Masaru、Ozaki Norio、Okano Hideyuki
  • Journal Title: eNEURO Volume: 6 Issue: 5 Pages: 0403-18
  • DOI: 10.1523/eneuro.0403-18.2019
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Rapid, robust, and subtype-specific neuronal differentiation from human pluripotent stem cells for multi-dimensional screening of neurodevelopmental / psychiatric disorders (2020)
  • Author(s): Ishikawa M
  • Organizer: The 10th Takeda Science Foundation Symposium on PharmaSciences
  • Int'l Joint Research
• [Presentation] Cellular Modeling of a Neurodevelopmental Disorder using Functionally Matured Human Excitatory / Inhibitory Neurons (2019)
  • Author(s): Ishikawa M and Okano H
  • Organizer: Neuro2019
  • Invited
• [Presentation] iPS細胞を利用した創薬の未来を語ろう：疾患iPS細胞を使って選抜した薬物でのALSの医師主導治験 (2019)
  • Author(s): 石川充
  • Organizer: 第10回 スクリーニング学研究会
  • Invited