Controlling of organ morphology by anchor proteins in regenerative organogenesis

• Project/Area Number: 18K19634
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 57:Oral science and related fields
• Research Institution: Tohoku University
• Principal Investigator: Nakamura Takashi 東北大学, 歯学研究科, 准教授 (90585324)
• Co-Investigator(Kenkyū-buntansha): 中村 はな 東邦大学, 医学部, 研究員 (30385827)
• Project Period (FY): 2018-06-29 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2019: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2018: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
• Keywords: エナメル芽細胞 / ギャップジャンクション / エピプロフィン / Single cell RNA-seq / 器官形成 / マイクロRNA / 歯の発生 / 組織再生 / 自己組織化 / 発生 / 再生医療 / Single-cell RNAシークエンス

Outline of Final Research Achievements

In the present study, we constructed a single-cell RNA sequence library from developing mouse tooth germ. We performed the rebuild the data on computer-based organ mapping, and performed individual gene expression profiling of dental epithelial cells. As a result, we succeeded in categorizing a variety of dental epithelial cells, and focus on the expression of gap junction protein connexin 43 (Cx43). We found that microRNA-1 (miR-1) is involved in Cx43 protein expression, and miR-1 inhibits Cx43 translation during tooth development and negatively regulates the proliferation of dental epithelial cells. Furthermore, the negative regulation in cell proliferation was associated to ATP release by the formation of hemi-channel Cx43 in differentiating dental epithelial cells with the modification of the cellular localization of Cx43.

Academic Significance and Societal Importance of the Research Achievements

本研究では、自己組織化する細胞集団からsingle-cell RNAシークエンスライブラリーを構築し、得られたデータをコンピューター上で器官マッピング構築後、マッピング構築されたSingle-cell RNAシークエンスデータを解析し、細胞外基質蛋白とそれらのインテグリンなどの受容体、細胞接着因子、細胞間結合蛋白、細胞増殖や分化誘導に関わる分泌系増殖因子とそれらの受容体に注目し、構造的アンカー分子と組織分化のトリガー分子の同定を目指す。 再生した器官が機能するに十分な大きさ、形態を兼ね備えさらには組織構成細胞のマッピングが適切に管理する技術があれば、より高度な器官再生医療が可能となる。

Research Products

• [Journal Article] Regulation of miR-1-Mediated Connexin 43 Expression and Cell Proliferation in Dental Epithelial Cells. (2020)
  • Author(s): Nakamura T, Iwamoto T, Nakamura HM, Shindo Y, Saito K, Yamada A, Yamada Y, Fukumoto S, Nakamura T.
  • Journal Title: Front Cell Dev Biol. Volume: 17(8) Pages: 156-156
  • DOI: 10.3389/fcell.2020.00156
  • NAID: 120006869515
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The transcription factor AmeloD stimulates epithelial cell motility essential for tooth morphology. (2019)
  • Author(s): Chiba Y, He B, Yoshizaki K, Rhodes C, Ishijima M, Bleck CKE, Stempinski E, Chu EY, Nakamura T, Iwamoto T, de Vega S, Saito K, Fukumoto S, Yamada Y
  • Journal Title: J Biol Chem Volume: 294(10) Issue: 10 Pages: 3406-3418
  • DOI: 10.1074/jbc.ra118.005298
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] The G-protein coupled receptor Gpr115 is essential for enamel mineralization via regulation of pH homeostasis (2019)
  • Author(s): Yuta Chiba, Keigo Yoshizaki, Tomoko Ikeuchi, Kan Saito, Craig Rhodes, Tsutomu Iwamoto, Takashi Nakamura, Yoshihiko Yamada and Satoshi Fukumoto
  • Organizer: NIH-Japan-JSPS symposium 2019
  • Int'l Joint Research