Role of intracellular osteopontin in the osteoclast fusion and tumor cell invasion

• Project/Area Number: 17591950
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Showa University
• Principal Investigator: SUZUKI Keiko Showa University, Sch. of Dent., assistant professor (50119187)
• Co-Investigator(Kenkyū-buntansha): SAKAI Nobuhiro Showa University, Sch. of Dent., Assistant Professor (90286849) ; YAMADA Shoji Showa University, Sch. of Dent., Professor (00111617) ; KAMATA Nobuyuki Hiroshima University, Grad. Sch. Biomed Sci., professor (70242211)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,580,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥180,000); Fiscal Year 2007: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2006: ¥600,000 (Direct Cost: ¥600,000); Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: intracellular osteopontin / osteoclast / cell fusion / squamous cell carcinoma / tumor cell invasion / cell migration / epithelial-mesenchymal transition / confocal laser-scanning microscopy / OsteoPontin / 細胞骨格再構築 / オステオポンティン / 細胞骨格

Research Abstract

Osteoclasts (OC) are the multinucleated cells which is responsible for the bone resorption in both physiological and pathological bone metabolism. Precursors of OC need to migrate and fuse together to form the multinucleated mature cells, which is an important step in OC differentiation. That osteopontin (OPN) has a prominent role in bone resorption was first indicated by the demonstration that it colocalizes with the avβ3 integrin and mediates the attachment of OC to the bone. However, it is still unclear at which stages of OC differentiation and activation that OPN functions. To clarify this, we examined the OC generated from bone marrow mononuclear precursors by laser scanning confocal microscopy. In prefusion OC and in motile multinucleated OC, staining for OPN colocalized with CD44 and β3 integrin, which are the OPN receptors, in cell processes. OPN was also found to colocalize with Cdc42 and Racl in filopodia and ruffled membranes, respectively. Immunocytochemical analysis of OC derived from OPN-null and CD44-null showed diminished perimembranous staining for CD44 and OPN, respectively, indicating a mutual interdependency of these proteins for their localization at cell surfaces. Moreover, OC generated from these mutant mice showed reduced cell spreading, limited protrusion of pseudopodia and impaired cell fusion compared to wild-type controls, consistent with the putative role of the OPN and CD44 in mediating cytoskeletal rearrangements. These studies indicate that OPN colocalizes with CD44 within osteoclast cell processes, and that OPN may be important in regulating CD44-mediated cytoskeletal changes associated with OC migration, fusion and resorption. Furthermore, it was confirmed that OPN played a critical role in wound healing process in squamous carcimona cells, since the front-line cells which protrude pseudopods and migrate toward the space between cell layers expressed OPN and CD44 strongly.

Research Products

• [Journal Article] Up-regulation of stromal cell-derived factor-1alpha and its receptor CXCR4 expression accompanied with epithelial-mesenchymal transition in human oral squamous cell carcinoma (2008)
  • Author(s): Taki M, Higashikawa K, Yoneda S, Ono S, Shigeishi H, Nagayama M, Kamata N
  • Journal Title: Oncol Reports 19(4) Pages: 993-998
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Gene expression profiling to identify genes associated with high-invasiveness in human squamous cell carcinoma with epithelial-to-mesenchymal transition (2008)
  • Author(s): Higashikawa K, Yoneda S, Taki M, Shigeishi H, Ono S, Tobiume K, Kamata N
  • Journal Title: Cancer Letter Mar7(Epub ahead of print)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Gene expression profiling to identify genes associated with high-invasiveness in human squamous cell carcinoma with epithelial-to-mesenchymal transition (2008)
  • Author(s): Higashikawa, K, Yoneda, S, Taki, M, Shigeishi, H, Ono, S, Tobiume, K, Kamata, N
  • Journal Title: Cancer Lett (Epub ahead of print)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Up-regulation of stromal cell-derived factor-lalpha and its receptor CXCR4 expression accompanied with epithelial-mesenchymal transition in human oral squamous cell carcinoma (2008)
  • Author(s): Taki, M, Higashikawa, K, Yoneda, S, Ono, S, Shigeishi, H, Nagayama, M, Kamata, N
  • Journal Title: Oncol Rep 19(4) Pages: 993-998
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Concrescence of teeth : cemental union between the crown of an impacted tooth and the roots of an erupted tooth (2007)
  • Author(s): Sugiyama, M, Ogawa, I, Suei, Y, Tohmori, H, Higashikawa, K, Kamata, N
  • Journal Title: J Oral Pathol Med 36(1) Pages: 60-62
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular characterization of GDD1/TMEM16E, the gene product responsible for autosomal dominant gnathodiaphyseal dysplasia (2007)
  • Author(s): Mizuta, K, Tsutsumi, S, Inoue, H, Sakamoto, Y, Miyatake, K, Miyawaki, K, Noji, S, Kamata, N, Itakura, M
  • Journal Title: BBRC 357(1) Pages: 126-132
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Snail-induced down-regulation of DeltaNp63alpha acquires invasive phenotype of human squamous cell carcinoma (2007)
  • Author(s): Higashikawa, K, Yoneda, S, Tobiume, K, Taki, M, Shigeishi, H, Kamata, N
  • Journal Title: Cancer Res 67(19) Pages: 9207-9213
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Interaction of E2F-Rb family members with corepressors binding to the adjacent E2F site. (2007)
  • Author(s): Nakajima, Y, Yamada, S, Kamata, N, Ikeda, MA
  • Journal Title: BBRC 364(4) Pages: 1050-1055
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Functional significance of intracellular form of osteopontin in the migration and fusion of osteoclasts (2007)
  • Author(s): Suzuki K., Yamada S
  • Journal Title: Dentistry in Japan 43 Pages: 150-153
  • NAID: 10021953675
• [Journal Article] Correlation of human Bubl expression with tumor-proliferating activity in salivary gland tumors (2006)
  • Author(s): Shigeishi, H, Yoneda, S, Taki, M, Nobumori, T, Ohta, K, Higashikawa, K, Yasui, W, Kamata, N
  • Journal Title: Oncol Rep 15(4) Pages: 933-938
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic analyses in patients with familial isolated hyperparathyroidism and hyperparathyroidism-jaw tumour syndrome (2006)
  • Author(s): Mizusawa, N, Uchino, S, Iwata, T, Tsuyuguchi, M, Suzuki, Y, Mizukoshi, T, Yamashita, Y, Sakurai, A, Suzuki, S, Beniko, M, Tahara, H, Fujisawa, M, Kamata, N, Fujisawa, K, Yashiro, T, Nagao, D, Golam, HM, Sano, T, Noguchi, S, Yoshimoto, K
  • Journal Title: Clin Endocrinol(Oxf) 65(1) Pages: 9-16
  • NAID: 120006631416
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Increased expression of CENP-H gene in human oral squamous cell carcinomas harboring high-proliferative activity (2006)
  • Author(s): Shigeishi, H, Higashikawa, K, Ono, S, Mizuta, K, Ninomiya, Y, Yoneda, S, Taki, M, Kamata, N
  • Journal Title: Oncol Rep 16(5) Pages: 1071-1075
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibitory effects of bisphosphonates on the differentiation osteoclasts (2006)
  • Author(s): K Suzuki
  • Journal Title: J Pharmacol Sci 100 Pages: 189-194
• [Journal Article] Risedronate Does Not Interfere with Bone Formation Lipopolysaccharide-Stimulated Bone Resorption (2006)
  • Author(s): K Suzuki
  • Journal Title: Dentistry in Japan 42 Pages: 38-42
• [Journal Article] Inhibitory effects of bisphosphonates on the differentiation and activity of osteoclasts. (2006)
  • Author(s): K Suzuki
  • Journal Title: J Pharmacol Sci 100 Pages: 189-194
• [Journal Article] Risedronate Does Not Interfere with Bone Formation Coupled with Lipopolysaccharide-Stimulated Bone Resorption. (2006)
  • Author(s): K Suzuki
  • Journal Title: Dentistry in Japan 42 Pages: 38-42
  • NAID: 10019595348
• [Journal Article] Osteoclast Responses to Lipopolysaccharide, Parathyroid Hormone and Bisphosphonates in Neonatal Murine Calvaria Analyzed by Laser Scanning Confocal Microscopy. (2005)
  • Author(s): K Suzuki
  • Journal Title: J Histochem Cytochem 53/12 Pages: 1525-1537
• [Journal Article] Exogenous Osteopontin Fails to Restore the Impaired Cell Spreading and Migration Observed in OPN-null Osteoclasts (2005)
  • Author(s): K Suzuki
  • Journal Title: Oral Ther Pharmacol 24/3 Pages: 87-94
  • NAID: 10017098144
• [Journal Article] オステオポンチンの遺伝子、構造、生合成. (2005)
  • Author(s): 鈴木 恵子
  • Journal Title: 日本臨床 63/10 Pages: 608-612
• [Journal Article] Pharmacological topics of bone metabolosm:A novel bisphosphonate for the treatment of periodontitis
  • Author(s): H.Shinoda, S.Takeyama, K.Suzuki, S.Murakami and S.Yamada
  • Journal Title: J Pharmacol.Sci
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Pharmacological topics of bone metabolosm : A novel bisphosphonate for the treatment of periodontitis
  • Author(s): Hisashi, Shinoda, Sadaaki, Takeyama, Keiko, Suzuki, Shinobu, Murakami, Shoji, Yamada
  • Journal Title: J Pharmacol. Sci (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Pharmacological topics of bone metabolosm:A novel bisphosphonate for the treatment of periodontitis
  • Author(s): H. Shinoda, S. Takeyama, K. Suzuki, S. Murakami, S. Yamada
  • Journal Title: J Pharmacol. Sci. (in press)
  • Peer Reviewed
• [Journal Article] Racl and Cdc42,members of Rho-family GTPase,have different subcellular distributions in osteoclasts
  • Author(s): Suzuki K., Yamada S
  • Journal Title: Oral Ther.Pharmacol. (in press)
  • Peer Reviewed
• [Journal Article] Functional significance of intracellular form of osteopontin in the migration and fusion of osteoclasts
  • Author(s): K Suzuki
  • Journal Title: Dentistry in Japan in press
  • NAID: 10021953675
• [Presentation] Pam_3CSK_4, a TLR2 agonist, induces osteoclastogenesis RANKL-independently (2008)
  • Author(s): Keiko, Suzuki, Akiyoshi, Hoshino, Kenji, Yamamoto, Shoji, Yamada
  • Organizer: The 81st Annual Meeting of The Japanese Pharmacological Society
  • Place of Presentation: Yokohama
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Intracellular Form of Osteopontin Plays an Important Role in Osteoclastogenesis and Bone Resorption (2007)
  • Author(s): K.Suzuki, SR.Rittling, DT.Denhardt, SYamada and J.Sodek
  • Organizer: Gordon Research Conference
  • Place of Presentation: Biddeford,USA
  • Year and Date: 2007-08-07
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Intracellular Form of Osteopontin Plays an Important Role in Osteoclastogenesis and Bone Resorption (2007)
  • Author(s): K. Suzuki, SR. Rittling, DT. Denhardt., S. Yamada, J. Sodek
  • Organizer: Gordon Research Conference
  • Place of Presentation: Biddeford.USA
  • Year and Date: 2007-08-07
• [Presentation] γ-glutamyltranspeptidase(GGT), a marker of inflammation, induces osteoclastogenesis by stimulating cytoskeletal rearrangement of preosteoclasts (2007)
  • Author(s): Keiko, Suzuki, Sawako, Moriwaki, Shumpei, Niida, Shoji, Yamada
  • Organizer: The 80th Annual Meeting of The Japanese Pharmacological Society
  • Place of Presentation: Nagoya
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] A novel bisphosphonate for the treatment of periodontitis (2007)
  • Author(s): Hisashi, Shinoda, Sadaaki, Takeyama, Keiko, Suzuki, Shinobu, Murakami, Shoji, Yamada
  • Organizer: The 80th Annual Meeting of The Japanese Pharmacological Society
  • Place of Presentation: Nagoya
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Relationship between bone resorption and bone formation in neonatal calvaria cultured with bisphosphonates (2007)
  • Author(s): Keiko, Suzuki, Sadaaki, Takeyama, Shinobu, Murakami, Hisashi, Shinoda, Shoji, Yamada
  • Organizer: The 25th Annual Meeting of The Japanese Society for Bone and Mineral Research
  • Place of Presentation: Osaka
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Jaro Sodek Intracellular Form of Osteopontin Plays an Important Role in Osteoclastogenesis and Bone Resorption (2007)
  • Author(s): Keiko, Suzuki, Susan, R., Rittling, David, T., Denhardt, Shoji, Yamada
  • Organizer: Gordon Research Conference
  • Place of Presentation: Biddeford, USA
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] GGT gamma -Glutamyltranspeptidase) Stimulates Osteoclast formation by Acting Directly on Osteoclast Precursor Cells (2007)
  • Author(s): S., Moriwaki, K., Suzuki, M., Takami, K., Ikeda, S., Niida
  • Organizer: 29th Annual Meeting of the American Society for Bone and Mineral Research
  • Place of Presentation: Hawaii, USA
  • Description: 「研究成果報告書概要(欧文)」より