Analysis of early post-implantation development and establishment of physiological ex vivo culture method in primate

• Project Area: A new foundation for primate developmental biology
• Project/Area Number: 20H05761
• Research Category: Grant-in-Aid for Transformative Research Areas (B)
• Allocation Type: Single-year Grants
• Review Section: Transformative Research Areas, Section (III)
• Research Institution: Kyoto University
• Principal Investigator: Nakamura Tomonori 京都大学, 白眉センター, 特定准教授 (90648429)
• Co-Investigator(Kenkyū-buntansha): 岡本 郁弘 京都大学, 高等研究院, 特定講師 (40648424)
• Project Period (FY): 2020-10-02 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥38,220,000 (Direct Cost: ¥29,400,000、Indirect Cost: ¥8,820,000); Fiscal Year 2022: ¥11,310,000 (Direct Cost: ¥8,700,000、Indirect Cost: ¥2,610,000); Fiscal Year 2021: ¥12,350,000 (Direct Cost: ¥9,500,000、Indirect Cost: ¥2,850,000); Fiscal Year 2020: ¥14,560,000 (Direct Cost: ¥11,200,000、Indirect Cost: ¥3,360,000)
• Keywords: 霊長類 / 発生学 / scRNA-seq / 着床後胚発生 / 次元の呪い / カニクイザル

Outline of Research at the Start

本研究計画A01では、研究領域の目標である非ヒト霊長類を用いた着床後胚の発生学研究基盤構築のうち、霊長類着床後胚発生研究が滞っていた大きな原因の一つである試料採取の困難性に対する回避策の提供を目標とする。このため、着床期のカニクイザル胚に出現する全細胞種をscRNA-seqにより同定し、同時に主要マーカー遺伝子の発現部位を明らかにすることで、位置情報を保持した遺伝子発現動態の完全解明を目指す。また生存率の極めて低い既存のEx vivo culture法を改良し、子宮内で発生する胚周囲の環境を参考に生存率向上を目指す。そしてA02で再現される子宮内膜環境への着床モデル構築を目指す。

Outline of Final Research Achievements

In this research project, we focused on primate-specific developmental phenomena immediately after implantation using cynomolgus macaques. First, we performed scRNA-seq on embryos at E15-E23 to identify all cell types emerging during early post-implantation. To this end, we developed an algorithm, RECODE, to solve the "curse of dimensionality" problem inherent in scRNA-seq. We found that the developmental stages of embryos change significantly between E17 and E19, and the characters of EXMC and AmEcto, which are not present in mice of the same age. We also found that the differences of inactivation mechanisms of the X chromosome between mice and the mokyes. Furthermore, we investigated the germline lineage induction from primate ESCs and found that the lineage is via AmEcto.

Academic Significance and Societal Importance of the Research Achievements

ヒトの流産の多くが初期に起こると言われているが、ヒトの着床直後の胚発生はなぞに包まれている。また多能性幹細胞を用いた再生医療/創薬研究も大きな期待を寄せられているが、霊長類多能性幹細胞の実態は未知な部分が多い。本研究成果は、大きな期待を寄せられている今後のヒト胚発生研究や臨床応用研究、さらには多能性幹細胞研究のための重要な参照データとなり、今後の研究発展を支える基盤となるものである。

Research Products

• [Journal Article] GATA transcription factors, SOX17 and TFAP2C, drive the human germ-cell specification program (2021)
  • Author(s): Kojima Yoji、Yamashiro Chika、Murase Yusuke、Yabuta Yukihiro、Okamoto Ikuhiro、Iwatani Chizuru、Tsuchiya Hideaki、Nakaya Masataka、Tsukiyama Tomoyuki、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Life Science Alliance Volume: 4 Issue: 5 Pages: 202000974-202000974
  • DOI: 10.26508/lsa.202000974
  • NAID: 120006979739
  • Peer Reviewed / Open Access
• [Journal Article] Capturing human trophoblast development with naive pluripotent stem cells in vitro (2021)
  • Author(s): S Io M Kabata Y Lemura N Morone A Minagawa B Wong I Okamoto T Nakamura Y Kojima C Iwatani H Tsuchiya B Kawswandy E KondohS Kaneko K Woltjen M Saitou T Yamanoto M Mandai Y Takashima
  • Journal Title: Cell Stem Cell Volume: 21 Issue: 6 Pages: 10-10
  • DOI: 10.1016/j.stem.2021.03.013
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cyclosporin A and FGF signaling support the proliferation/survival of mouse primordial germ cell-like cells in vitro† (2020)
  • Author(s): Ohta Hiroshi、Yabuta Yukihiro、Kurimoto Kazuki、Nakamura Tomonori、Murase Yusuke、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Biology of Reproduction Volume: 104 Issue: 2 Pages: 344-360
  • DOI: 10.1093/biolre/ioaa195
  • Peer Reviewed / Open Access
• [Journal Article] ZGLP1 is a determinant for the oogenic fate in mice (2020)
  • Author(s): Nagaoka So I.、Nakaki Fumio、Miyauchi Hidetaka、Nosaka Yoshiaki、Ohta Hiroshi、Yabuta Yukihiro、Kurimoto Kazuki、Hayashi Katsuhiko、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Science Volume: 367 Issue: 6482 Pages: 4115-4115
  • DOI: 10.1126/science.aaw4115
  • NAID: 120006850276
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Long‐term expansion with germline potential of human primordial germ cell‐like cells in?vitro (2020)
  • Author(s): Murase Yusuke、Yabuta Yukihiro、Ohta Hiroshi、Yamashiro Chika、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: The EMBO Journal Volume: 39 Issue: 21 Pages: 104929-104929
  • DOI: 10.15252/embj.2020104929
  • Peer Reviewed
• [Presentation] 霊長類特異的多能性状態の解明 (2020)
  • Author(s): Tomonori Nakamura* , Yusuke Imoto, Yasuaki Hiraoka, Mitinori Saitou
  • Organizer: 第53回発生生物学会年会
  • Int'l Joint Research / Invited
• [Presentation] Identifying cells hidden by curse of dimensionality (2020)
  • Author(s): Tomonori Nakamura
  • Organizer: The 1st ASHBi SignAC international workshop 2021
  • Int'l Joint Research / Invited
• [Patent(Industrial Property Rights)] 観測ノイズ削減法の開発 (2020)
  • Inventor(s): 井元祐介、中村友紀、平岡裕章、斎藤通紀
  • Industrial Property Rights Holder: 井元祐介、中村友紀、平岡裕章、斎藤通紀
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020