Elucidation of the role of miRNA on the control of cellular function by ultrasound

• Project/Area Number: 20K12619
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: University of Toyama
• Principal Investigator: Tabuchi Yoshiaki 富山大学, 学術研究部薬学・和漢系, 教授 (20322109)
• Co-Investigator(Kenkyū-buntansha): 長谷川 英之 富山大学, 学術研究部工学系, 教授 (00344698) ; 鈴木 信雄 金沢大学, 環日本海域環境研究センター, 教授 (60242476) ; 古澤 之裕 富山県立大学, 工学部, 准教授 (80632306) ; 平山 順 公立小松大学, 保健医療学部, 教授 (90510363)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 超音波 / 細胞分化 / アクチン / マイクロRNA / 石灰化 / 遺伝子発現 / 熱作用 / miRNA

Outline of Research at the Start

低出力パルス超音波 (LIPUS) は，臨床で骨折治療の補助療法として用いられている．しかしながら，この安全で有効なLIPUSの作用の分子メカニズムは不明な点が多い．マイクロRNA (miRNA) は，遺伝子の発現を調節し様々な生命現象に関与することから，医療応用が期待されている．本申請では，LIPUSによる細胞の分化や増殖等の機能制御においてmiRNAがどの様に関与しているかを，骨芽細胞等で検討する．LIPUSのmiRNAに関連した分子メカニズムが明らかになれば，LIPUSの骨折治癒促進作用機序の解明や新規骨疾患治療への貢献に繋がる．

Outline of Final Research Achievements

In order to elucidate the cellular response to low-intensity pulsed ultrasound (LIPUS), the effects of LIPUS on the osteogenic differentiation in osteoblastic cells were investigated. Mouse osteoblastic MC3T3-E1 cells were exposed to LIPUS (30 mW/cm2 for 20 min; once a day). Repeated exposure of LIPUS significantly elevated mineralization level detected using alizarin red staining. Morphological changes were observed in the cells treated with siRNA for actin beta. The silencing of actin beta diminished the osteoblastic differentiation that was increased in the MC3T3-E1 cells by exposure to LIPUS. These findings will provide a molecular basis for further understanding of the mechanisms of LIPUS in osteoblastic differentiation.

Academic Significance and Societal Importance of the Research Achievements

低出力パルス超音波 (LIPUS) は，骨折治癒促進効果があるがその分子機構は完全に解明されていない．LIPUSの細胞応答を明らかにするために，骨芽細胞の骨分化に対するLIPUSの効果を検討した．マウス骨芽MC3T3-E1細胞へのLIPUSの連続照射は，骨分化を有意に上昇させ，この作用の少なくとも一部にアクチンベータが関与することが判った．得られた成績は， LIPUSの作用メカニズムを理解するための重要な分子基盤になる．

Research Products

• [Journal Article] Hydroxylated benzo[c]phenanthrene metabolites cause osteoblast apoptosis and skeletal abnormalities in fish (2022)
  • Author(s): Suzuki Nobuo, Honda Masato, Sekiguchi Toshio, Toriba Akira, Tang Ning, Edward Nagato Go, Zhang Lulu, Hayakawa Kazuichi, et all.
  • Journal Title: Ecotoxicology and Environmental Safety Volume: 234 Pages: 113401-113401
  • DOI: 10.1016/j.ecoenv.2022.113401
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Osteoclastic and Osteoblastic Responses to Hypergravity and Microgravity: Analysis Using Goldfish Scales as a Bone Model (2022)
  • Author(s): Yamamoto Tatsuki、Ikegame Mika、Furusawa Yukihiro、Tabuchi Yoshiaki、Hatano Kaito、Watanabe Kazuki、Kawago Umi、Hirayama Jun、Yano Sachiko、Sekiguchi Toshio、Kitamura Kei-ichiro、Endo Masato、Nagami Arata、Matsubara Hajime、Maruyama Yusuke、Hattori Atsuhiko、Suzuki Nobuo
  • Journal Title: Zoological Science Volume: 39 Issue: 4 Pages: 388-396
  • DOI: 10.2108/zs210107
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Activation of RANKL-producing cells under simulated microgravity with a three-dimensional clinostat in regenerating goldfish scales (2022)
  • Author(s): Yamamoto, T., Ikegame, M., Kuroda, K., Kobayashi-Sun, J., Hirayama, J., Kobayashi, I., Kawamura, R., Endo, M., Tabuchi, Y., Furusawa, Y., Yachiguchi, K., Sekiguchi, T., Matsubara, H., Yano, S., Hattori, A. and Suzuki, N.
  • Journal Title: Biological Sciences in Space Volume: 36 Issue: 0 Pages: 9-14
  • DOI: 10.2187/bss.36.9
  • ISSN: 0914-9201, 1349-967X
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Glyoxal‐induced formation of advanced glycation end‐products in type 1 collagen decreases both its strength and flexibility in vitro (2021)
  • Author(s): Kitamura Kei‐ichiro、Hirayama Jun、Tabuchi Yoshiaki、Minami Takao、Matsubara Hajime、Hattori Atsuhiko、Suzuki Nobuo
  • Journal Title: Journal of Diabetes Investigation Volume: 12 Issue: 9 Pages: 1555-1559
  • DOI: 10.1111/jdi.13528
  • Peer Reviewed / Open Access
• [Journal Article] Neurotoxicity of a pyrethroid pesticide deltamethrin is associated with the imbalance in proteolytic systems caused by mitophagy activation and proteasome inhibition (2021)
  • Author(s): Hirano Tetsushi、Suzuki Nihei、Ikenaka Yoshinori、Hoshi Nobuhiko、Tabuchi Yoshiaki
  • Journal Title: Toxicology and Applied Pharmacology Volume: 430 Pages: 115723-115723
  • DOI: 10.1016/j.taap.2021.115723
  • Peer Reviewed / Open Access
• [Journal Article] Low-intensity ultrasound inhibits melanoma cell proliferation in vitro and tumor growth in vivo (2021)
  • Author(s): Feril Loreto B.、Yamaguchi Kazuki、Ikeda-Dantsuji Yurika、Furusawa Yukihiro、Tabuchi Yoshiaki、Takasaki Ichiro、Ogawa Ryohei、Cui Zheng-Guo、Tachibana Katsuro
  • Journal Title: Journal of Medical Ultrasonics Volume: 48 Issue: 4 Pages: 451-461
  • DOI: 10.1007/s10396-021-01131-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genetic response to low-intensity ultrasound on mouse ST2 bone marrow stromal cells (2021)
  • Author(s): Tabuchi Yoshiaki、Hasegawa Hideyuki、Suzuki Nobuo、Furusawa Yukihiro、Hirano Tetsushi、Nagaoka Ryo、Hirayama Jun、Hoshi Nobuhiko、Mochizuki Takashi
  • Journal Title: Molecular Medicine Reports Volume: 23 Issue: 3 Pages: 173-173
  • DOI: 10.3892/mmr.2020.11812
  • Peer Reviewed / Open Access
• [Journal Article] Expression of sclerostin in the regenerating scales of goldfish and its increase under microgravity during space flight (2020)
  • Author(s): Yamamoto, T., Ikegame, M., Hirayama, J., Kitamura, K., Tabuchi, Y., Furusawa, Y., Sekiguchi, T., Endo, M., Mishima, H., Seki, A., Yano, S., Matsubara H., Hattori, A. and Suzuki, N.
  • Journal Title: Biomedical Research Volume: 41 Issue: 6 Pages: 279-288
  • DOI: 10.2220/biomedres.41.279
  • NAID: 130007948844
  • ISSN: 0388-6107, 1880-313X
  • Year and Date: 2020-12-01
  • Peer Reviewed / Open Access
• [Journal Article] Detection of RANKL-producing cells and osteoclastic activation by the addition of exogenous RANKL in the regenerating scales of goldfish. (2020)
  • Author(s): Yamamoto, T., Ikegame, M., Kawago, U., Tabuchi, Y., Hirayama, J., Sekiguchi, T., Furusawa, Y., Yachiguchi, K., Matsubara, H., Urata, M., Hattori, A. and Suzuki, N.
  • Journal Title: Biological Science in Space Volume: 34 Issue: 0 Pages: 34-40
  • DOI: 10.2187/bss.34.34
  • NAID: 130007884615
  • Peer Reviewed / Open Access
• [Presentation] マウス骨由来細胞における骨形成分化に対する低出力パルス超音波の効果 (2022)
  • Author(s): 田渕圭章，平野哲史，長岡 亮，長谷川英之
  • Organizer: 日本超音波医学会第95回学術集会
• [Presentation] 哺乳動物細胞における低出力パルス超音波誘導遺伝子発現に対する熱作用の関与の検討 (2021)
  • Author(s): 田渕圭章，長谷川英之，望月 剛
  • Organizer: 日本超音波医学会第94回学術集会
  • Invited
• [Presentation] 骨髄間質細胞における低出力パルス超音波に対する初期応答遺伝子群の同定 (2020)
  • Author(s): 田渕圭章，長谷川英之，鈴木信雄，平野哲史，長岡 亮，望月 剛
  • Organizer: 日本超音波医学会第93回学術集会