メチローム解析による試験管内始原生殖細胞誘導過程における遺伝子発現制御機構の解明

• Project/Area Number: 14J04310
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Developmental biology
• Research Institution: Kyushu University
• Principal Investigator: 白根 健次郎 九州大学, 医学系学府, 特別研究員(DC1)
• Project Period (FY): 2014-04-25 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 2016: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2015: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2014: ¥1,000,000 (Direct Cost: ¥1,000,000)
• Keywords: DNAメチル化 / 始原生殖細胞 / 多能性幹細胞 / 高速シークエンサー / ヒストン修飾 / リプログラミング / エピゲノム / 次世代シークエンサー

Outline of Annual Research Achievements

本研究は胚性幹細胞からエピブラスト様細胞を経て誘導された始原生殖細胞様細胞をモデルとして生殖細胞形成過程におけるDNAメチル化制御機構と遺伝子発現とメチル化の関係性を明らかにすることを目的としている。本年度は昨年度までに取得したデータの詳細な解析を行い、胚性幹細胞と始原生殖細胞様細胞におけるDNAメチル化による遺伝子発現制御は大きく異なることを発見した。胚性幹細胞では多能性の維持に関わる遺伝子を含む遺伝子周辺は低いDNAメチル化状態であり、転写活性化と関わるのに対して、始原生殖細胞様細胞ではポリコームのターゲット遺伝子である発生関連遺伝子周囲のDNA が低いメチル化状態となるもののH3K27me3によりその抑制状態を保つことがわかった。また、胚性幹細胞と始原生殖細胞様細胞における低いDNAのメチル化状態の確立と維持にはPrdm14という因子が必須であることを突き止めた。更に、メスの胚性幹細胞、エピブラスト様細胞、始原生殖細胞様細胞のDNAメチル化レベルはオスと比べて低いこと、またメスの胚性幹細胞とエピブラスト様細胞では特定のがん細胞に特徴的なラミナ会合ドメインにおけるメチル化の低下が見られた。

Research Progress Status

28年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

28年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Software updates in the Illumina HiSeq platform affect whole-genome bisulfite sequencing (2017)
  • Author(s): Toh, H., Shirane, K. (co-first author), Miura, F., Kubo, N., Ichiyanagi, K., Hayashi, K., Saitou, M., Suyama, M., Ito, T. & Sasaki, H
  • Journal Title: BMC Genomics Volume: 18 Issue: 1 Pages: 31-38
  • DOI: 10.1186/s12864-016-3392-9
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Global Landscape and Regulatory Principles of DNA Methylation Reprogramming for Germ Cell Specification by Mouse Pluripotent Stem Cells (2016)
  • Author(s): Shirane K, Kurimoto K, Yabuta Y, Yamaji M, Satoh J, Ito S, Watanabe A, Hayashi K, Saitou M, Sasaki H
  • Journal Title: Dev Cell Volume: 39 Issue: 1 Pages: 87-103
  • DOI: 10.1016/j.devcel.2016.08.008
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Allele-specific methylome and transcriptome analysis reveals widespread imprinting in the human placenta (2016)
  • Author(s): Hamada, H., Okae, H., Toh, H., Chiba, H., Hiura, H., Shirane, K., Sato, T., Suyama, M., Yaegashi, N., Sasaki, H. & Arima
  • Journal Title: The American Journal of Human Genetics Volume: 99 Issue: 5 Pages: 1045-1058
  • DOI: 10.1016/j.ajhg.2016.08.021
  • Peer Reviewed / Open Access
• [Journal Article] In Vitro Derivation and Propagation of Spermatogonial Stem Cell Activity from Mouse Pluripotent Stem Cells (2016)
  • Author(s): Ishikura Y, Yabuta Y, Ohta H, Hayashi K, Nakamura T, Okamoto I, Yamamoto T, Kurimoto K, Shirane K, Sasaki H, Saitou M
  • Journal Title: Cell Rep Volume: 17 Issue: 10 Pages: 2789-2804
  • DOI: 10.1016/j.celrep.2016.11.026
  • Peer Reviewed / Open Access
• [Journal Article] De novo DNA methylation drives 5hmC accumulation in mouse zygotes. (2016)
  • Author(s): Amouroux R, Nashun B, Shirane K, Nakagawa S, Hill PW, D'Souza Z, Nakayama M, Matsuda M, Turp A, Ndjetehe E, Encheva V, Kudo NR, Koseki H, Sasaki H, Hajkova P.
  • Journal Title: Nat. Cell Biol. Volume: 18 Issue: 2 Pages: 225-233
  • DOI: 10.1038/ncb3296
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] DNA methylation and gene expression dynamics during spermatogonial stem cell differentiation in the early postnatal mouse testis. (2015)
  • Author(s): Kubo N, Toh H, Shirane K, Shirakawa T, Kobayashi H, Sato T, Sone H, Sato Y, Tomizawa S, Tsurusaki Y, Shibata H, Saitsu H, Suzuki Y, Matsumoto N, Suyama M, Kono T, Ohbo K, Sasaki H.
  • Journal Title: BMC Genomics. Volume: 16 Issue: 1 Pages: 624-624
  • DOI: 10.1186/s12864-015-1833-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Analysis of influenza A/H3N2 neuraminidase genes obtained from influenza patients in the 2011/12 and 2012/13 seasons in Japan (2015)
  • Author(s): Ikematsu, H., Chong, Y., Shirane, K., Toh, H., Sasaki, H., Koga, Y., Matsumoto, S., Hotta, T., Uchiumi, T., Kang, D.
  • Journal Title: Fukuoka Acta Medica Volume: 106 Pages: 16-22
  • Peer Reviewed
• [Journal Article] Analysis of the Neuraminidase Amino Acid Sequences of Influenza A/H1N1pdm09, A/H3N2, and B Viruses Isolated from Influenza Patients in the 2013/14 Japanese Influenza Season (2015)
  • Author(s): Ikematsu, H., Chong, Y., Shirane, K., Toh, H., Sasaki, H., Koga, Y., Matsumoto, S., Hotta, T., Uchiumi, T., Kang, D.
  • Journal Title: Fukuoka Acta Medica Volume: 106 Pages: 231-239
  • Peer Reviewed
• [Journal Article] Setdb1 is required for germ line development and silencing of H3K9me3 marked endogenous retroviruses in primordial germ cells. (2014)
  • Author(s): Liu, S., Brind’Amour, J., Karimi, M.M., Shirane, K., Bogutz, A., Lefebvre, L., Sasaki, H., Shinkai, Y. & Lorincz, M.C.
  • Journal Title: Genes and Devlopment Volume: 28 Issue: 18 Pages: 2041-2055
  • DOI: 10.1101/gad.244848.114
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Global landscape and regulatory principles of DNA methylation reprogramming for germ cell specification by mouse pluripotent stem cells (2016)
  • Author(s): Shirane, K., Kurimoto, K., Yabuta, Y., Yamaji, M., Satoh, J., Ito, S., Watanabe, A., Hayashi, K., Saitou, M. & Sasaki, H
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] Global landscape and regulatory principles of DNA methylation reprogramming for germ cell specification by mouse pluripotent stem cells (2016)
  • Author(s): Shirane, K., Kurimoto, K., Yabuta, Y., Yamaji, M., Satoh, J., Ito, S., Watanabe, A., Hayashi, K., Saitou, M. & Sasaki, H
  • Organizer: 第１０回日本エピジェネティクス研究会年会
  • Place of Presentation: 大阪
  • Year and Date: 2016-05-19
• [Presentation] Global landscape and regulatory principles of DNA methylation reprogramming for germ cell specification by mouse pluripotent stem cells (2016)
  • Author(s): Shirane, K., Kurimoto, K., Yabuta, Y., Yamaji, M., Satoh, J., Ito, S., Watanabe, A., Hayashi, K., Saitou, M. & Sasaki, H
  • Organizer: Cold Spring Harbor Asia meeting: Chromatin, Epigenetics and Transcription
  • Place of Presentation: 蘇州
  • Year and Date: 2016-05-09
  • Int'l Joint Research
• [Presentation] Global Landscape and Regulatory Principle of DNA Methylation Reprogramming for Germ Cell Specification by Mouse Pluripotent Stem Cells (2016)
  • Author(s): Kenjiro Shirane, Kazuki Kurimoto, Yukihiro Yabuta, Masashi Yamaji, Akira Watanabe, Katsuhiko Hayashi, Mitinori Saitou, Hiroyuki Sasaki
  • Organizer: Keystone Symposia Chromatin and Epigenetics Meeting
  • Place of Presentation: ウィスラー、カナダ
  • Year and Date: 2016-03-20
  • Int'l Joint Research
• [Presentation] Global Landscape and Regulatory Principle of DNA Methylation Reprogramming for Germ Cell Specification by Mouse Pluripotent Stem Cells (2016)
  • Author(s): Kenjiro Shirane, Kazuki Kurimoto, Yukihiro Yabuta, Masashi Yamaji, Akira Watanabe, Katsuhiko Hayashi, Mitinori Saitou, Hiroyuki Sasaki
  • Organizer: Epigenome Dynamics and Regulation in Germ Cells
  • Place of Presentation: 京都
  • Year and Date: 2016-02-17
  • Int'l Joint Research
• [Presentation] 体外誘導系マウス始原生殖細胞分化過程のDNAメチル化動態と遺伝子発現制御 (2015)
  • Author(s): 白根健次郎、栗本一基、薮田幸宏、林克彦、大田浩、中村友紀、渡辺亮、斎藤通紀、佐々木裕之
  • Organizer: 第３８回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] 体外誘導系マウス始原生殖細胞分化過程のDNAメチル化動態と遺伝子発現制御 (2015)
  • Author(s): 白根健次郎、栗本一基、林克彦、渡辺亮、斎藤通紀、佐々木裕之
  • Organizer: 第９回日本エピジェネティクス研究会年会
  • Place of Presentation: 東京一ツ橋学術総合センター
  • Year and Date: 2015-05-25 – 2015-05-26
• [Presentation] 体外誘導系マウス始原生殖細胞分化過程のDNAメチル化動態と遺伝子発現制御 (2015)
  • Author(s): 白根健次郎、栗本一基、林克彦、渡辺亮、斎藤通紀、佐々木裕之
  • Organizer: 第９回日本エピジェネティクス研究会年会
  • Place of Presentation: 東京
  • Year and Date: 2015-05-25
• [Presentation] 体外誘導系から見えてきた始原生殖細胞分化過程のDNAメチル化動態と遺伝子発現制御 (2014)
  • Author(s): 白根健次郎、栗本一基、林克彦、渡辺亮、斎藤通紀、佐々木裕之
  • Organizer: 第17回生体防御医学研究所リトリート
  • Place of Presentation: ホテルセキア
  • Year and Date: 2014-07-22 – 2014-07-23
• [Presentation] 試験管内始原生殖細胞誘導における全ゲノムDNAメチル化解析 (2014)
  • Author(s): 白根健次郎、栗本一基、林克彦、渡辺亮、斎藤通紀、佐々木裕之
  • Organizer: 生殖細胞のエピゲノムダイナミクスとその制御若手勉強会
  • Place of Presentation: つくばグランドホテル
  • Year and Date: 2014-07-16 – 2014-07-18