Elucidation of the molecular mechanism in the osteoclast function through protein kinase N3 and its clinical application.

• Project/Area Number: 19K10050
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57010:Oral biological science-related
• Research Institution: Matsumoto Dental University
• Principal Investigator: Uehara Shunsuke 松本歯科大学, 歯学部, 講師 (90434480)
• Co-Investigator(Kenkyū-buntansha): 小林 泰浩 松本歯科大学, 総合歯科医学研究所, 教授 (20264252)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 破骨細胞 / 骨吸収 / Pkn3 / 骨粗鬆症

Outline of Research at the Start

本研究の目的は、1）In vitro及びin vivoにおけるPkn3阻害剤の骨吸収に対する効果、2）Pkn3によるc-Src活性化の分子機構、を明らかにすることである。
そのために、培養破骨細胞の骨吸収活性に対するPkn3阻害剤の作用の解明、卵巣切除モデル（骨吸収亢進により骨量減少を呈する）マウスの骨量に対するPkn3阻害剤の治療効果の検証、Pkn3によりリン酸化され、c-Srcの活性化に関与する分子の同定を行う。

Outline of Final Research Achievements

In osteoporosis and inflammatory bone diseases, excessive bone resorption results in loss of bone mass. In order to prevent the deterioration of bone formation due to the failure of osteoclast-osteoblast coupling, it is necessary to develop a drug that suppresses only the bone-resorbing activity of osteoclasts. We found Protein kinase N3 (Pkn3), which enhances the function of osteoclasts downstream of the Wnt signaling. Since this protein is rarely expressed in normal tissues other than osteoclasts, it is suitable as a target for developing drugs with few side effects. In this research project, we administered a low-molecular-weight compound that had been reported to have a Pkn3 inhibitory effect on model mice showing bone loss due to increased bone resorption. This compound alleviated bone loss by suppressing bone resorption.

Academic Significance and Societal Importance of the Research Achievements

骨粗鬆症は、日本国内だけでも推定1000万人以上の患者さんが存在する疾患である。骨量の減少による、骨折の増加は、QOLを低下させるだけでなく、寝たきりにつながる可能性もある。骨量の減少は、骨吸収を行う破骨細胞の働きが、骨形成を行う骨芽細胞の働きを上回ることで生じるため、破骨細胞の骨吸収機能を抑制する化合物は、骨粗鬆症の治療薬になり得る。我々は、Pkn3というタンパク質が破骨細胞機能に重要であることを見出した。本研究はその阻害剤が、骨量減少モデルマウスの治療に利用できることを示したもので、臨床応用へつながる成果である。

Research Products

• [Journal Article] Inhibitor of protein kinase N3 suppresses excessive bone resorption in ovariectomized mice (2022)
  • Author(s): Shunsuke Uehara, Hideyuki Mukai, Teruhito Yamashita, Masanori Koide, Kohei Murakami, Nobuyuki Udagawa, Yasuhiro Kobayashi
  • Journal Title: J Bone Miner Metab . Volume: 40 Issue: 2 Pages: 251-261
  • DOI: 10.1007/s00774-021-01296-1
  • Peer Reviewed / Open Access
• [Journal Article] Characterization, pharmacokinetics, and pharmacodynamics of anti-Siglec-15 antibody and its potency for treating osteoporosis and as follow-up treatment after parathyroid hormone use. (2021)
  • Author(s): Tsuda E, Fukuda C, Okada A, Karibe T, Hiruma Y, Takagi N, Isumi Y, Yamamoto T, Hasegawa T, Uehara S, Koide M, Udagawa N, Amizuka N, Kumakura S.
  • Journal Title: Bone. Volume: Epub 2021 Oct 27. Pages: 116241-116241
  • DOI: 10.1016/j.bone.2021.116241
  • Peer Reviewed / Open Access
• [Journal Article] Osteoclast differentiation by RANKL and OPG signaling pathways (2020)
  • Author(s): Udagawa Nobuyuki、Koide Masanori、Nakamura Midori、Nakamichi Yuko、Yamashita Teruhito、Uehara Shunsuke、Kobayashi Yasuhiro、Furuya Yuriko、Yasuda Hisataka、Fukuda Chie、Tsuda Eisuke
  • Journal Title: Journal of Bone and Mineral Metabolism Volume: 39 Issue: 1 Pages: 19-26
  • DOI: 10.1007/s00774-020-01162-6
  • Peer Reviewed / Open Access
• [Journal Article] The Vitamin D Receptor in Osteoblast-Lineage Cells Is Essential for the Proresorptive Activity of 1α,25(OH)2D3 In Vivo (2020)
  • Author(s): Tomoki Mori, Kanji Horibe, Masanori Koide, Shunsuke Uehara, Yoko Yamamoto, Shigeaki Kato, Hisataka Yasuda, Naoyuki Takahashi, Nobuyuki Udagawa, Yuko Nakamichi
  • Journal Title: Endocrinology . Volume: 161 Issue: 11 Pages: 1-14
  • DOI: 10.1210/endocr/bqaa178
  • Peer Reviewed / Open Access
• [Journal Article] Sclerostin expression in trabecular bone is downregulated by osteoclasts (2020)
  • Author(s): Masanori Koide, Teruhito Yamashita, Kohei Murakami, Shunsuke Uehara, Keigo Nakamura, Midori Nakamura, Mai Matsushita, Toshiaki Ara, Hisataka Yasuda, Josef M Penninger, Naoyuki Takahashi, Nobuyuki Udagawa, Yasuhiro Kobayashi
  • Journal Title: Sci Rep . Volume: 10 Issue: 1 Pages: 13751-13751
  • DOI: 10.1038/s41598-020-70817-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulations of osteoclast formation and function by Wnt signals. (2019)
  • Author(s): Kobayashi Y, Uehara S, Koide M
  • Journal Title: Clin Calcium Volume: 29 Pages: 309-315
• [Journal Article] Regulation of osteoclast function via Rho-Pkn3-c-Src pathways. (2019)
  • Author(s): Uehara S, Udagawa N, Kobayashi Y
  • Journal Title: J Oral Biosci Volume: 61 Issue: 3 Pages: 135-140
  • DOI: 10.1016/j.job.2019.07.002
  • Peer Reviewed
• [Journal Article] PKN1 kinase-negative knock-in mice develop splenomegaly and leukopenia at advanced age without obvious autoimmune-like phenotypes. (2019)
  • Author(s): Siddique SM, Kubouchi K, Shinmichi Y, Sawada N, Sugiura R, Itoh Y, Uehara S, Nishimura K, Okamura S, Ohsaki H, Kamoshida S, Yamashita Y, Tamura S, Sonoki T, Matsuoka H, Itoh T, Mukai H.
  • Journal Title: Sci Rep. Volume: 27;9(1) Issue: 1 Pages: 13977-13993
  • DOI: 10.1038/s41598-019-50419-2
  • NAID: 120006733136
  • Peer Reviewed / Open Access
• [Journal Article] The Regulation of Bone Metabolism and Disorders by Wnt Signaling. (2019)
  • Author(s): Maeda K, Kobayashi Y, Koide M, Uehara S, Okamoto M, Ishihara A, Kayama T, Saito M, Marumo K
  • Journal Title: Int J Mol Sci Volume: 20 Issue: 22 Pages: 5525-5525
  • DOI: 10.3390/ijms20225525
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] プロテインキナーゼN3 (Pkn3) 阻害剤は、卵巣切除に伴う骨量減少を骨吸収抑制により軽減する (2020)
  • Author(s): 上原俊介，山下照仁，村上康平，小出雅則，宇田川信之，小林泰浩
  • Organizer: 第38回日本骨代謝学会学術集会