Establishment of high efficient chromosomal aneuploidy rescue method by genome editing in iPS cells

• Project/Area Number: 19K16520
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 48040:Medical biochemistry-related
• Research Institution: Hiroshima University
• Principal Investigator: Akutsu Silvia Natsuko 広島大学, 原爆放射線医科学研究所, 助教 (10822299)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: Trisomy rescue / iPSC / chromosome / cytogenetics / trisomy / rescue / トリソミー・レスキュー / 染色体 / iPS細胞 / iPS cells / Chromosome / Aneuploidy / Reprogramming / Down syndrome / ゲノム編集

Outline of Research at the Start

ダウン症候群は、トリソミー21を原因とする最も患者数の多い染色体異数性病であるが、その根治的な治療法は未だに確立されていない。最近、iPS細胞への初期化でトリソミー21がレスキューされる「染色体異数性治療法」の可能性が報告された。本研究では、ゲノム編集技術を用いて、ダウン症候群iPS細胞における１）マウス2細胞期発現遺伝子および山中4因子の均一な誘導発現系、２）2細胞期(2C)様細胞集団の濃縮技術を構築して、高効率な染色体異数性レスキュー実験系を確立することを研究目的としている。本研究成果は、ダウン症候群の根治療法に向けた画期的な基盤技術となると期待される。

Outline of Final Research Achievements

I obtained dermal fibroblasts from eight patients with major human trisomy syndromes (trisomy 21, 18, 13, and 9 syndromes), established 190 clones of iPS cells, and comprehensively examined the copy number of genomic DNA in all clones. The number of copies of genomic DNA in all clones was detailed examined by karyotype, FISH, SNP and Sanger sequencing. As a result, the trisomy rescue phenomenon was confirmed in at least one cell line of each trisomy syndrome. Chromosome loss occurred regardless of parental origin. These results suggest that when pluripotency is induced by iPS cell reprogramming, 1) one of the trisomy chromosomes is randomly lost from the cells, and 2) cells normalized to disomy are sorted to form single iPS cell colonies. Furthermore, late lagging was postulated as the mechanism of chromosome loss.

Academic Significance and Societal Importance of the Research Achievements

トリソミーレスキューは、着床前の初期胚でも観察されており、染色体数を正確に保つ生体のメカニズムの一つと考えられます。iPS細胞のトリソミーレスキューは、初期胚のトリソミーレスキューとメカニズムを共有する可能性があることから、本研究成果は生殖補助医療への貢献が期待されます。また、iPS細胞リプログラミングによるトリソミーレスキューは、ゲノム操作を伴わない染色体を修正する新たな治療法として、不妊症やがん治療などの再生・移植医療への応用が期待されます。

Research Products

• [Journal Article] iPSC reprogramming-mediated aneuploidy correction in autosomal trisomy syndromes (2022)
  • Author(s): Akutsu Silvia Natsuko、Miyamoto Tatsuo、Oba Daiju、Tomioka Keita、Ochiai Hiroshi、Ohashi Hirofumi、Matsuura Shinya
  • Journal Title: PLOS ONE Volume: 17 Issue: 3 Pages: 1-20
  • DOI: 10.1371/journal.pone.0264965
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Imaging of the Ciliary Cholesterol Underlying the Sonic Hedgehog Signal Transduction (2022)
  • Author(s): Miyamoto T, Hosoba K, Akutsu SN, Matsuura S
  • Journal Title: Methods Mol Biol Volume: 2374 Pages: 49-57
  • DOI: 10.1007/978-1-0716-1701-4_5
  • ISBN: 9781071617007, 9781071617014
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] NBS1 I171V variant underlies individual differences in chromosomal radiosensitivity within human populations (2021)
  • Author(s): Tomioka K, Miyamoto T, Akutsu SN, Yanagihara H, Fujita K, Royba E, Tauchi H, Yamamoto T, Koh I, Hirata E, Kudo Y, Kobayashi M, Okada S, Matsuura S
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 1-13
  • DOI: 10.1038/s41598-021-98673-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Premature aging syndrome showing random chromosome number instabilities with CDC20 mutation (2020)
  • Author(s): Fujita Harumi、Sasaki Takashi、Miyamoto Tatsuo、Akutsu Silvia Natsuko、Sato Showbu、Mori Takehiko、Nakabayashi Kazuhiko、Hata Kenichiro、Suzuki Hisato、Kosaki Kenjiro、Matsuura Shinya、Matsubara Yoichi、Amagai Masayuki、Kubo Akiharu
  • Journal Title: Aging Cell Volume: 19
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Insufficiency of ciliary cholesterol in hereditary Zellweger syndrome (2020)
  • Author(s): Miyamoto Tatsuo、Hosoba Kosuke、Itabashi Takeshi、Iwane Atsuko H、Akutsu Silvia Natsuko、Ochiai Hiroshi、Saito Yumiko、Yamamoto Takashi、Matsuura Shinya
  • Journal Title: The EMBO Journal Volume: 39
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Quantitative evaluation of the NBS1 I171V variant as a candidate genetic factor underlying individual differences in radiation sensitivity (2020)
  • Author(s): Tomioka K, Fujita K, Akutsu SN, Yanagihara H, Tauchi H, Yamamoto T, Kobayashi M, Kudo Y, Miyamoto T, Matsuura S
  • Journal Title: 広島医学 Volume: 73 Pages: 224-227
  • Open Access
• [Journal Article] Applications of Genome Editing Technology in Research on Chromosome Aneuploidy Disorders (2020)
  • Author(s): Akutsu Silvia Natsuko、Fujita Kazumasa、Tomioka Keita、Miyamoto Tatsuo、Matsuura Shinya
  • Journal Title: Cells Volume: 9 Issue: 1 Pages: 239-239
  • DOI: 10.3390/cells9010239
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] iPSC reprogramming-mediated random trisomy correction in aneuploidy syndromes (2022)
  • Author(s): Silvia Natsuko Akutsu
  • Organizer: The 6th International Symposium of the Network-type Joint Usage/ Research Center for Radiation Disaster Medical Science
  • Int'l Joint Research
• [Presentation] トリソミー症候群の細胞初期化で誘導されるトリソミーレスキュー (2021)
  • Author(s): Silvia Natsuko Akutsu
  • Organizer: The 64th Annual Meeting of the Japanese Radiation Research Society,
• [Presentation] トリソミー症候群の細胞初期化で誘導されるトリソミーレスキュー (2021)
  • Author(s): Silvia Natsuko Akutsu, Tatsuo Miyamoto, Keita Tomioka, Daiju Oba, Hirofumi Ohashi, Shinya Matsuura
  • Organizer: The 5th International Symposium of the Network-type Joint Usage/ Research Center for Radiation Disaster Medicine Science
  • Int'l Joint Research
• [Presentation] Correction of aneuploidy in human cell cultures derived from patients with autosomal trisomy syndromes (2021)
  • Author(s): Silvia Natsuko Akutsu
  • Organizer: 第６回HiHA young researchers
  • Invited
• [Presentation] iPSC reprogramming-mediated aneuploidy correction in autosomal trisomy syndromes. (2021)
  • Author(s): Akutsu SN, Miyamoto T, Tomioka K, Oba D, Ohashi H, Matsuura S.
  • Organizer: 放射線災害・医科学研究拠点 第5回国際シンポジウム
  • Int'l Joint Research
• [Presentation] Quantitative evaluation of the NBS1 I171V variant on radiosensitivity. (2020)
  • Author(s): Tomioka K, Fujita K, Akutsu SN, Tauchi H, Yamamoto T, Kobayashi M, Kudo Y, Miyamoto T, Matsuura S.
  • Organizer: 放射線災害・医科学研究拠点 第4回国際シンポジウム
  • Int'l Joint Research
• [Presentation] NBS1 I171V 多型による放射線感受性個人差の定量的評価 (2020)
  • Author(s): 冨岡啓太、Silvia Natsuko Akutsu、柳原啓見、田内 広、山本 卓、小林正夫、工藤美樹 、藤田和将、宮本達雄、松浦伸也
  • Organizer: 第４５回中国地区放射線影響研究会プログラム
• [Presentation] A novel premature aging syndrome showing random chromosome number instability with CDC20 mutation (2020)
  • Author(s): 藤田春美、佐々木貴史、宮本達雄、阿久津シルビア夏子、佐藤尚武、森毅彦、中林一彦、秦健一郎、鈴木寿人、小崎健次郎、松浦伸也、松原洋一、天谷雅行、久保亮治.
  • Organizer: 日本人類遺伝学会第65回大会
• [Presentation] Peroxisome biogenesis disorder Zellweger syndrome as a ciliopathy (2020)
  • Author(s): 宮本達雄、細羽康介、板橋岳志、岩根敦子、阿久津シルビア夏子、落合博、斎藤裕見子、山本卓、松浦伸也
  • Organizer: 日本人類遺伝学会第65回大会
• [Presentation] Peroxisomes ensure to supply cholesterol into the ciliary membrane : a lesson from a peroxisome-biogenesis disorder Zellweger syndrome. (2020)
  • Author(s): Miyamoto T, Hosoba K, Itabashi T, Iwane AH, Akutsu SN, Ochiai H, Saito Y, Yamamoto T, Matsuura S.
  • Organizer: 日本分子生物学会第４３回
• [Presentation] The generation of mosaic trisomy 21 model cells using patient cells with full trisomy 21 by trisomy rescue during cell reprogramming and their modification with fluorescent nuclear markers by genome editing technique (2019)
  • Author(s): Silvia Natsuko Akutsu
  • Organizer: The 65th Brazilian Congress of Genetics
  • Int'l Joint Research / Invited
• [Presentation] Generation of Down syndrome iPS cells tagged with fluorescence marker in chromosome 21 using genome editing technology (2019)
  • Author(s): Silvia Natsuko Akutsu
  • Organizer: 第64回大会 日本人類遺伝学会 （ポスター発表）
• [Presentation] CRISPR-ObLiGaRe法を用いたiPS細胞における蛍光核標識によるモザイク・トリソミー21のモデル細胞系の開発 (2019)
  • Author(s): Silvia Natsuko Akutsu
  • Organizer: The 4th Annual Meeting ofthe Japanese Society for Genome Editing（ポスター発表）
• [Remarks] iPSC reprogramming-mediated aneuploidy correction
  • URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0264965
• [Remarks] iPS細胞リプログラミングによるトリソミー染色体の自己修正
  • URL: https://www.hiroshima-u.ac.jp/news/69742