Study on extra-cellular function mediated by functional RNAs in exosomes secreted from pluripotent stem cells

• Project/Area Number: 18K09803
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57060:Surgical dentistry-related
• Research Institution: Osaka Dental University
• Principal Investigator: NOZAKI Tadashige 大阪歯科大学, 歯学部, 教授 (90281683)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 再生医学 / 幹細胞 / エクソソーム / microRNA / 次世代シークエンス

Outline of Final Research Achievements

Functional RNAs in exosomes secreted from cells participate in a variety of cell functions and are expected as biomarkers. Poly ADP-ribosylation is a post-translational modification that controls major functional and biological processes in cells, including production of exosomes. To determine the effect of poly ADP-ribosylation on functional RNAs in exosomes secreted from dental pulp stem cells and its ramifications on downstream signal transduction, we knocked down poly(ADP-ribose) polymerase 1 (PARP1) in dental pulp stem cells using genome editing or inhibitors and performed an exhaustive analysis of the expression of microRNAs derived from the exosomes and the resulting alterations in cell functions. We found that PARP1 could potentially control senescence mediated by their exosomes in dental pulp stem cells.

Academic Significance and Societal Importance of the Research Achievements

組織幹細胞である歯髄幹細胞は「再生医療等製品」として臨床への応用が期待されている。しかしながら、長期にわたり継代培養を続けると、細胞老化によって増殖能や分化能が低下するという問題点がある。ポリADP-リボシル化は翻訳後修飾の重要なファクターであり、老化に深く関与していると考えられている。本研究では、PARP1が歯髄幹細胞で産生されるエクソソームに含まれる機能性RNAを制御することで、細胞老化をコントロールしている可能性を見出した。PARP1がエクソソーム機能性RNAを介した細胞老化に関与することを示した本研究は、老化に関わる新たなメカニズムの解明に発展することが期待される。

Research Products

• [Journal Article] Spontaneous Development of Dental Dysplasia in Aged Parp-1 Knockout Mice (2019)
  • Author(s): Hisako Fujihara, Tadashige Nozaki, Masahiro Tsutsumi, Mayu Isumi, Shinji Shimoda, Yoshiki Hamada, Mitsuko Masutani
  • Journal Title: Cells Volume: 27 Issue: 10 Pages: 1157-1157
  • DOI: 10.3390/cells8101157
  • NAID: 120006988121
  • Peer Reviewed / Open Access
• [Journal Article] Development of renal failure in PargParp-1 null and Timm23 hypomorphic mice (2019)
  • Author(s): Chen Lichao、Gunji Akemi、Uemura Akiko、Fujihara Hisako、Nakamoto Kentaro、Onodera Takae、Sasaki Yuka、Imamichi Shoji、Isumi Mayu、Nozaki Tadashige、Kamada Nobuo、Jishage Kou-ichi、Masutani Mitsuko
  • Journal Title: Biochemical Pharmacology Volume: 167 Pages: 116-124
  • DOI: 10.1016/j.bcp.2019.07.003
  • Peer Reviewed / Open Access
• [Journal Article] Dysfunction of Poly (ADP-Ribose) Glycohydrolase Induces a Synthetic Lethal Effect in Dual Specificity Phosphatase 22-Deficient Lung Cancer Cells (2019)
  • Author(s): Sasaki Yuka、Fujimori Hiroaki、Hozumi Miyuki、Onodera Takae、Nozaki Tadashige、Murakami Yasufumi、Ashizawa Kazuto、Inoue Kengo、Koizumi Fumiaki、Masutani Mitsuko
  • Journal Title: Cancer Research Volume: 79 Issue: 15 Pages: 3851-3861
  • DOI: 10.1158/0008-5472.can-18-1037
  • Peer Reviewed / Open Access
• [Journal Article] Measurement of Poly(ADP-ribose) Level with Enhanced Slot Blot Assay with Crosslinking (2018)
  • Author(s): Kudo Yuko、Sasaki Yuka、Onodera Takae、Hashimoto Jun、Nozaki Tadashige、Tamura Kenji、Watanabe Masatoshi、Masutani Mitsuko
  • Journal Title: Challenges Volume: 9 Issue: 2 Pages: 27-27
  • DOI: 10.3390/challe9020027
  • Peer Reviewed / Open Access