低分子天然化合物を用いた骨形成の治療標的分子の同定

• Project/Area Number: 11F01113
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 外国
• Research Field: Morphological basic dentistry
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 山口 朗 東京医科歯科大学, 大学院・医歯学総合研究科, 教授
• Co-Investigator(Kenkyū-buntansha): LEE J.-W. 東京医科歯科大学, 大学院・医歯学総合研究科, 外国人特別研究員
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2012: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2011: ¥800,000 (Direct Cost: ¥800,000)
• Keywords: 骨芽細胞 / 破骨細胞 / 低分子化合物 / 口腔癌 / CXCL2 / RANKL / 低分子天延化合物 / 骨形成 / honokiol / 骨破壊

Research Abstract

本研究では、以下の成果を得た。
1)Acer nikoenseの骨芽細胞に及ぼす作用:Acer nikoenseから分離した6種類の化合物を検討し、diphenyl ether-type cyclic diarylheptanoidsが骨芽細胞分化を促進することを明らかにした。
2)Rosmarinic acidの骨芽細胞と破骨細胞に及ぼす作用:Rosmarinic acidは、古典的Wnt経路を介して骨芽細胞分化を促進するとともに、活性型ビタミンDやRANKLで誘導された破骨細胞形成を抑制した。破骨細胞の抑制過程では、RANKLの誘導するp38のリン酸化とRANKLの誘導するNFATc1, c-Jun, c-Fosの転写活性を抑制することに起因することを明らかにした。
3)口腔癌による骨破壊メカニズム:
A:口腔癌の産生するCXCL2の役割の解析:口腔癌細胞株HSC3より十数の単一細胞に由来するクローン化細胞株を樹立し、各細胞株がST2細胞のRANKL発現に及ぼす作用とヌードマウス頭蓋部移植による骨破壊能を検討した。その結果、HSC3-C13はHSC3-C17に比べてRANKL誘導脳と骨破壊能が顕著に高いことを見いだした。そのため、両細胞間での遺伝子発現をマイクロアレイで解析したところ、HSC3-C13ではCXCL2の発現が顕著であった。そして、CXCL2は口腔癌の産生する破骨細胞性骨吸収促進因子であることを明らかにした。
B:骨破壊における口腔癌細胞と間質の産生するRANKLの役割の区別化:ヒト口腔癌細胞をヌードマウスの頭蓋部に移植する骨破壊モデルを用いた。この場合、癌細胞の産生するRANKLはヒト由来で、間質の産生するRANKLはマウス由来である。ヒトとマウスのRANKLを特異的に中和するRANKL抗体を用いて、癌細胞および間質の産生するRANKLが口腔癌の骨破壊で重要であることを明らかにした。

Research Products

• [Journal Article] RANKL synthesized by both stromal cells and cancer cells plays a crucial role in osteoclastic bone resorption induced by oral cancer (2013)
  • Author(s): Kiyoshi Sato, Ji-Won Lee, Kei Sakamoto, Tadahiro Iimura, Kou Kayamori, Hisataka Yasuda, Masanobu Shindoh, Masako Ito, Ken Omura, Akira Yamaguchi
  • Journal Title: American Journal of Pathology Volume: 182 Issue: 5 Pages: 1890-1899
  • DOI: 10.1016/j.ajpath.2013.01.038
  • Peer Reviewed
• [Journal Article] Effects of veraguensin and galgravin on osteoclast differentiation and function (2012)
  • Author(s): Asai M, Ji-Won Lee, Itakura Y, Choi BK, Yonezawa T, Teruya T, Cha BY, Ohnishi M, Yamaguchi A, Woo JT
  • Journal Title: Cytotechnology Volume: 64 Issue: 3 Pages: 315-322
  • DOI: 10.1007/s10616-011-9416-z
  • Peer Reviewed
• [Journal Article] CxCL2 synthesized by oral squamous cell carcinoma is involved in cancer-associated bone destruction (2012)
  • Author(s): Oue E, Lee Ji-Won, Sakamoto K, Iimura K, Aoki K, Kayamori K, Michi Y, Yamagshiro M, Harada K, Amagasa T, Yamaguchi A
  • Journal Title: Biochem Biophys Res Commun Volume: 424 Issue: 3 Pages: 456-461
  • DOI: 10.1016/j.bbrc.2012.06.132
  • Peer Reviewed
• [Journal Article] Down-regulation of keratin 4 and keratin 13 expression in oral squamous cell carcinoma and epithelial dysplasia : a clue for histopathogenesis (2011)
  • Author(s): Sakamoto K, Aragaki T, Kawachi H, Katsube K, Miki Y, Takizawa T, Omura K, Morita K, Okada N, Yamaguchi A
  • Journal Title: Histopathol Volume: 58 Pages: 531-542
  • Peer Reviewed
• [Journal Article] Acerogenin A, a natural compound isolated from Acer nikoense Maxim, stimulates osteoblast differentiation through bone morphogenetic protein action (2011)
  • Author(s): Kihara K, Ichikawa S, Yonezawa T, Lee JW, Akihisa T, Woo JT, Michi Y, Amagasa T, Yamaguchi A
  • Journal Title: Biochem Bioph Res Commun Volume: 406 Issue: 2 Pages: 211-217
  • DOI: 10.1016/j.bbrc.2011.02.017
  • Peer Reviewed
• [Journal Article] Comparative morphology of osteocytes in aquatic and land vertebrates (2011)
  • Author(s): Cao L, Moriishi T, Miyazaki T, Iimura T, Tamamura S, Komori T, Yamaguchi A
  • Journal Title: J Bone Miner Metab Volume: 29 Issue: 6 Pages: 662-670
  • DOI: 10.1007/s00774-011-0268-6
  • Peer Reviewed
• [Journal Article] TGF-β derives epithelial-mesenchymal transition through δEF1-mediated downregulation of ESRP (2011)
  • Author(s): 堀口華奈
  • Journal Title: Oncogene Volume: (掲載確定)(印刷中) Issue: 26 Pages: 3190-3201
  • DOI: 10.1038/onc.2011.493
  • Peer Reviewed
• [Journal Article] Structural differences in the osteocyte networ between the calvaria and long bone revealed by three-dimensional fluorescence morphometry, possibly reflecting dstinct mechano-adaptations and sensitivities (2011)
  • Author(s): Himeno-Ando A, Izumi Y, Yamaguchi A, Iimura T
  • Journal Title: Biochem Bioph Res Commun Volume: 417 Issue: 2 Pages: 765-770
  • DOI: 10.1016/j.bbrc.2011.12.031
  • Peer Reviewed
• [Journal Article] Reduction of NOTCH1 expression pertains to maturation abnormalities of keratinocytes in squamous neoplasms (2011)
  • Author(s): Sakamoto K, Aragaki T, Kawachi H, Katsube K, Miki Y, Takizawa T, Omura K, Morita K, Okada N, Yamaguchi A
  • Journal Title: Lob Invest Volume: 92 Issue: 4 Pages: 688-702
  • DOI: 10.1111/j.1365-2559.2011.03759.x
  • Peer Reviewed
• [Journal Article] Expression of basal cell keratin 15 and keratin 19 in oral squamous cell carcinoma represent diverse phthophysiologies (2011)
  • Author(s): Khanom R, Sakamoto K, Pal SK, Shimada Y, Morita K-i, Omura K, Miki Y, Yamaguchi A
  • Journal Title: Histol Histopathol Volume: (In press)
  • Peer Reviewed
• [Journal Article] Canine oral mucosal fibroblasts differentiate into osteoblastic cellsin response to BMP-2 (2011)
  • Author(s): Umenara K, Iimura T, Sakamoto K, Lin Z, Kasugai S, Igarashi Y, Yamaguchi A
  • Journal Title: Anat Rec Volume: (In press)
  • Peer Reviewed
• [Journal Article] Clinical significance of lymphatic and blood vessel invasion in oral tongue squamous cell carcinomas (2011)
  • Author(s): Michikawa C, Uzawa N, Kayamori K, Sonoda I, Ohyama Y, Okada N, Yamaguchhi A, Amagasa T
  • Journal Title: Oral Oncol Volume: 48 Issue: 4 Pages: 320-324
  • DOI: 10.1016/j.oraloncology.2011.11.014
  • Peer Reviewed
• [Journal Article] Cdc42 is required for chondrogenesis and interdigital programmed cell death during limb development (2011)
  • Author(s): Aizawa R, Yamada A, Suzuki D, Iimura T, Kassai H, Harada T, Tsukasaki M, Yamamoto G, Tachikawa T, Nakao K, Yamamoto M, Yamagchi A, Aiba A, Kamijo R
  • Journal Title: AMech Dev Volume: (In press) Issue: 1-4 Pages: 38-50
  • DOI: 10.1016/j.mod.2012.02.002
  • Peer Reviewed
• [Journal Article] Ectopic production of hair keratin constitutes Rushton's hyaline bodies in association with hematogenous deposits° (2011)
  • Author(s): Sakamoto K, Khanom R, Hamagaki M, Yamaguchi A
  • Journal Title: J Oral Pathol Med Volume: (In press) Issue: 8 Pages: 637-641
  • DOI: 10.1111/j.1600-0714.2012.01150.x
  • Peer Reviewed
• [Presentation] 口腔癌の骨破壊 (2012)
  • Author(s): 山口朗
  • Organizer: 第30回日本口腔腫瘍学会
  • Place of Presentation: 大宮(教育講演)
  • Year and Date: 2012-01-27
• [Presentation] 腔癌による骨破壊の分子メカニズム (2011)
  • Author(s): 山口朗
  • Organizer: 第56回日本口腔外科学会総会
  • Place of Presentation: 大阪(シンポジウム)
  • Year and Date: 2011-10-22
• [Presentation] Histopathological characteristics of bisphosphonate-related osteomyelitis (osteonecrosis) of jaw (2011)
  • Author(s): Akira Yamaguchi
  • Organizer: 5^<th> Meeting of Asian Society of Oral and Maxillofacial Pathology
  • Place of Presentation: 福岡(Keynote Lecture)
  • Year and Date: 2011-08-30
• [Presentation] Molecular mechanism of bone destruction by oral cancer (2011)
  • Author(s): Yamaguchi A
  • Organizer: The Second Chinese National Conference on Oral Maxillofacial Development and Regeneration
  • Place of Presentation: Wuyishan city, Fujian Province, China
  • Year and Date: 2011-07-29
• [Presentation] 血管石灰化と骨・軟骨関連因子 (2011)
  • Author(s): 山口朗
  • Organizer: 第56回日本透析医学会学術総会
  • Place of Presentation: 横浜市(シンポジウム)
  • Year and Date: 2011-06-18
• [Presentation] ビスフォスフォネート関連顎骨壊死(骨髄炎)の病理学的所見 (2011)
  • Author(s): 山口朗
  • Organizer: 第31回日本骨形態計測学会
  • Place of Presentation: 岐阜市(シンポジウム)
  • Year and Date: 2011-05-20
• [Presentation] 骨再生の分子基盤 (2011)
  • Author(s): 山口朗
  • Organizer: 第65回日本口腔科学会学術集会
  • Place of Presentation: 東京(シンポジウム)
  • Year and Date: 2011-04-15
• [Presentation] ビスホスホネート関連顎骨壊死(骨髄炎) (2011)
  • Author(s): 山口朗
  • Organizer: 第65回日本口腔科学会学術集会
  • Place of Presentation: 東京(シンポジウム)
  • Year and Date: 2011-04-14
• [Presentation] 再生医療「骨再生」-そのメカニズムと応用- (2010)
  • Author(s): 山口朗
  • Organizer: 平成22年日本歯科医師会生涯研修セミナー
  • Place of Presentation: 津市
  • Year and Date: 2010-06-27