Elucidation of the mechanism inducing genome instability in human pluripotent stem cells and development of novel test method to assess genome instability

• Project/Area Number: 17K01443
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Medical engineering assessment
• Research Institution: National Institute of Health Sciences
• Principal Investigator: Miura Takumi 国立医薬品食品衛生研究所, 再生・細胞医療製品部, 室長 (60405355)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
• Keywords: ヒト多能性幹細胞 / ゲノム不安定性 / 品質評価 / レギュラトリーサイエンス / 再生医療 / 次世代シーケンサー

Outline of Final Research Achievements

It has been known that human pluripotent stem cells undergo genomic instability due to artificial stimuli such as continuous culture and differentiation induction. In this study, in order to ensure the safety of human pluripotent stem cells that can be used for clinical applications, we aimed to clarify the difference between "harmless genetic mutations" and "potentially harmful genetic mutations". We detected the DNA mutations that arose during cell culture and showed that some mutations directly or indirectly might confer a selective growth advantage to the cell.

Academic Significance and Societal Importance of the Research Achievements

再生医療への応用が期待されているヒト多能性幹細胞において、そのゲノム不安定性に寄与する遺伝子変異を同定することで、これまで未解明であった「ゲノム変異」と「造腫瘍性」との関連が明白になると期待される。つまり、「無害なゲノム異常」と「腫瘍を誘発するゲノム異常」との違いを明らかにすることは、ヒト多能性幹細胞由来製品の品質管理においても大いに役立ち、また、安全性を担保した培養環境を整備することも可能になる。

Research Products

• [Journal Article] A simple method to estimate the in-house limit of detection for genetic mutations with low allele frequencies in whole-exome sequencing analysis by next-generation sequencing (2021)
  • Author(s): Takumi Miura、Satoshi Yasuda、Yoji Sato
  • Journal Title: BMC Genomic Data Volume: 22 Issue: 1 Pages: 8-8
  • DOI: 10.1186/s12863-020-00956-x
  • Peer Reviewed / Open Access
• [Journal Article] The hsa-miR-302 cluster controls ectodermal differentiation of human pluripotent stem cell via repression of DAZAP2 (2020)
  • Author(s): Sugawara T, Miura T, Kawasaki T, Umezawa A, Akutsu H.
  • Journal Title: Regen Ther Volume: 15 Pages: 1-9
  • DOI: 10.1016/j.reth.2020.03.011
  • Peer Reviewed / Open Access
• [Journal Article] SALL3 expression balance underlies lineage biases in human induced pluripotent stem cell differentiation (2019)
  • Author(s): Kuroda Takuya、Yasuda Satoshi、Tachi Shiori、Matsuyama Satoko、Kusakawa Shinji、Tano Keiko、Miura Takumi、Matsuyama Akifumi、Sato Yoji
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 2175-2175
  • DOI: 10.1038/s41467-019-09511-4
  • Peer Reviewed / Open Access
• [Journal Article] Tumorigenicity-associated characteristics of human iPS cell lines (2018)
  • Author(s): Yasuda S, Kusakawa S, Kuroda T, Miura T, Tano K, Takada N, Matsuyama S, Matsuyama A, Nasu M, Umezawa A, Hayakawa T, Tsutsumi H, Sato Y.
  • Journal Title: PLOS ONE Volume: 13 Issue: 10 Pages: e0205022-e0205022
  • DOI: 10.1371/journal.pone.0205022
  • Peer Reviewed / Open Access
• [Journal Article] 米国における再生医療の規制の動向とヒトES細胞の医療応用の現状 (2017)
  • Author(s): 三浦 巧
  • Journal Title: 再生医療と医事法（医事法講座第8巻） Volume: 8 Pages: 121-134
• [Presentation] ヒトiPS細胞における神経分化予測マーカーの同定 (2021)
  • Author(s): 黒田 拓也，安田 智，松山 さと子，三浦 巧, 松山 晃文，川路 英哉，伊藤 昌可，河合 純，佐藤 陽治
  • Organizer: 第20回再生医療学会総会（Web開催）
• [Presentation] A transcriptomic approach for predictive markers to select human induced pluripotent stem cells with high neural progenitor cell differentiation potential (2020)
  • Author(s): Kuroda T, Yasuda S, Miura T, Itoh M, Kawaji H, Kawai J, Sato Y
  • Organizer: International Society for Stem Cell Research 2020 Annual Meeting Virtual
• [Presentation] ヒトiPS細胞分化傾向予測マーカーSALL3の機能解析 (2019)
  • Author(s): 黒田拓也，安田智，城しおり，松山さと子，草川森士，田埜慶子，三浦巧，松山晃文，佐藤陽治
  • Organizer: 日本再生医療学会 第1回秋季科学シンポジウム
• [Presentation] SALL3 expression balance underlies lineage biases in human iPS cell differentiation (2019)
  • Author(s): Kuroda T, Yasuda S, Tachi S, Matsuyama S, Kusakawa S, Tano K, Miura T, Matsuyama A, Sato Y
  • Organizer: International Society for Stem Cell Research 2019 Annual Meeting
  • Int'l Joint Research
• [Presentation] Searching for a predictive biomarker to select human induced pluripotent stem cells with high cardiac differentiation potential (2019)
  • Author(s): Ohashi F, Miyagawa S, Yasuda S, Miura T, Kuroda T, Itoh M, Kawaji H, Ito E, Yoshida S, Saito A, Oyama K, Matsuda I, Sameshima T, Kawai J, Sawa Y, Sato Y
  • Organizer: International Society for Stem Cell Research 2019 Annual Meeting
  • Int'l Joint Research
• [Presentation] 心筋細胞に分化指向性を有するiPS細胞株を選択するためのバイオマーカーの探索 (2019)
  • Author(s): 大橋文哉、宮川繁、安田 智、三浦 巧、黒田拓也、伊藤昌可、川路英哉、伊東絵望子、 吉田昇平、齋藤充弘、大山賢二、松田 勇、鮫島 正、河合 純、澤 芳樹、佐藤陽治
  • Organizer: 第18回日本再生医療学会総会
• [Presentation] 医療応用を目指したゲノム編集技術における品質・安全性評価の考え方 (2018)
  • Author(s): 三浦 巧、佐藤陽治
  • Organizer: 第17回 日本再生医療学会総会
  • Invited
• [Presentation] 多能性幹細胞加工製品中に混在する未分化性/造腫瘍性細胞の検出法の開発 (2018)
  • Author(s): 三浦 巧
  • Organizer: 第123回 日本解剖学会総会・全国学術集会
  • Invited
• [Presentation] ASSESSMENT OF THE APPEARANCE OF SPONTANEOUS GENEMIC MUTATIONS IN HUMAN INDUCED PLURIPOTENT STEM CELLS (2017)
  • Author(s): Miura T, Yasuda S, Okamura K, Kusakawa S, Umezawa A, Sato Y.
  • Organizer: International Society for Stem Cell Research 2017
  • Int'l Joint Research