Verification of the effect of De novo DNA methylation elimination on early embryogenesis of human ES/iPS cells.

• Project/Area Number: 20H03829
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 56040:Obstetrics and gynecology-related
• Research Institution: Tokai University
• Principal Investigator: Fukuda Atsushi 東海大学, 医学部, 講師 (00638091)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000); Fiscal Year 2022: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2021: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2020: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: エピジェネティクス / 初期発生 / 幹細胞生物学

Outline of Research at the Start

本研究では、ヒト多能性幹細胞における新規DNAメチル化酵素であるDNMT3A/3Bを抑制することにより、DNMT3A/3B欠損ヒト多能性幹細胞が、ヒトエピブラストにどの程度類似するかをトランスクリプトーム解析を通じて分子レベルで解明する。

Outline of Final Research Achievements

Human pluripotent stem cells (hPSCs) are in vitro products capable of differentiating into almost all the cells that make up the body. however, genome wide DNA methylation status of hPSCs are different compared to those of human preimplantation embryos (epiblast).
In this study, we asked how blocking of de novo DNA methyltransfereases activity in hPSCs affects transcriptome and epigenomic status. The deep sequencing analysis revealed that the effect of transcriptomic status in de novo DNA methyltransfereases deletion greatly depends on genetic background. Interestingly, we found that female cell specific abnormality, erosion of X-chromosome dosage compensation, is prevented by the mutations. Thus, our findings indicate that de novo DNA methyltransfereases activity is one of the drivers responsible for in vitro specific abnormality in hPSCs.

Academic Significance and Societal Importance of the Research Achievements

ヒト多能性幹細胞は、体を構成するほぼ全ての細胞に分化可能な細胞であり、再生・細胞医療において中心的な役割を担っている。近年は、特定の疾患由来のiPS細胞など、疾患ゲノムを考慮したGenotype-Phenotype解析にも活用され、創薬開発分野でも注目を浴びている。
ヒト多能性幹細胞は、ヒトの初期胚細胞と同様の機能を持つと推測されているが、試験管産物特有の異常も報告されている。本研究では、ヒト多能性幹細胞の試験管産物特有の現象を解明することで、より高品質なヒト多能性幹細胞の作製と応用的基盤の構築を目指すことを目的とした。

Research Products

• [Journal Article] Nanosheet coating improves stability of human pluripotent stem cell culture on glass substrates (2023)
  • Author(s): Sakata Yuka、Zhang Hong、Sugiyama Akiko、Motosugi Nami、Kimura Hiroshi、Okamura Yosuke、Fukuda Atsushi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 650 Pages: 55-61
  • DOI: 10.1016/j.bbrc.2023.01.077
  • Peer Reviewed / Open Access
• [Journal Article] De-erosion of X chromosome dosage compensation by the editing of XIST regulatory regions restores the differentiation potential in hPSCs (2022)
  • Author(s): Motosugi Nami、Sugiyama Akiko、Okada Chisa、Otomo Asako、Umezawa Akihiro、Akutsu Hidenori、Hadano Shinji、Fukuda Atsushi
  • Journal Title: Cell Reports Methods Volume: 2 Issue: 12 Pages: 100352-100352
  • DOI: 10.1016/j.crmeth.2022.100352
  • Peer Reviewed / Open Access
• [Journal Article] De novo DNA methyltransferases DNMT3A and DNMT3B are essential for XIST silencing for erosion of dosage compensation in pluripotent stem cells (2021)
  • Author(s): Fukuda Atsushi、Hazelbaker Dane Z.、Motosugi Nami、Hao Jin、Limone Francesco、Beccard Amanda、Mazzucato Patrizia、Messana Angelica、Okada Chisa、San Juan Irune Guerra、Qian Menglu、Umezawa Akihiro、Akutsu Hidenori、Barrett Lindy E.、Eggan Kevin
  • Journal Title: Stem Cell Reports Volume: 16 Issue: 9 Pages: 2138-2148
  • DOI: 10.1016/j.stemcr.2021.07.015
  • Peer Reviewed / Open Access / Int'l Joint Research