腫瘍の形成と維持の機構

• Project/Area Number: 12215024
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 渋谷 正史 東京大学, 医科学研究所, 教授 (10107427)
• Co-Investigator(Kenkyū-buntansha): 後藤 典子 東京大学, 医科学研究所, 講師 (10251448) ; 矢花 直幸 東京大学, 医科学研究所, 助手 (90292846) ; 櫻井 佳子 東京大学, 医科学研究所, 助手 (10361619) ; 大場 基 昭和大学, 腫瘍分子医学研究所, 助手 (70297018) ; 中村 佐千枝 東京大学, 医科学研究所, 助手 (60313087) ; 丸 義朗 東京大学, 医科学研究所, 助教授 (00251447)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥213,000,000 (Direct Cost: ¥213,000,000); Fiscal Year 2004: ¥60,000,000 (Direct Cost: ¥60,000,000); Fiscal Year 2003: ¥38,000,000 (Direct Cost: ¥38,000,000); Fiscal Year 2002: ¥38,000,000 (Direct Cost: ¥38,000,000); Fiscal Year 2001: ¥32,000,000 (Direct Cost: ¥32,000,000); Fiscal Year 2000: ¥45,000,000 (Direct Cost: ¥45,000,000)
• Keywords: 腫瘍血管 / VEGF / VEGF受容体 / ノックインマウス / シグナル伝達 / 血管新生 / VEGFR-2 / VEGFR-1 / がん転移 / 血管透過性 / VEGFR-2 (KDR) / VEGFR-1 (Flt-1) / VEGFR-2(KDR) / VEGFR-1(Flt-1) / 系統発生 / トランスジェニックマウス / VEGF-E

Research Abstract

VEGFとその受容体のシグナル伝達機構をマウス個体レベルで解析し、(1)内皮細胞増殖の主なポジティブシグナルを発信するのはVEGF受容体2(KDR/Flk-1)であること、また自己リン酸化部位1173チロシンは血管発生・新生に必須であること、を明らかにした。すなわち、1173チロシンをフェニールアラニンに置換したマウス(1173F/Fマウス)を作出して解析したところ、この変異マウスは血管系をほとんど構築することができず、胎生8.5-9.0日で死亡することが明らかとなった。この部位はPLCgamma-PKC-MAPキナーゼ経路の開始点であり、腫瘍血管系制御の新規分子標的として利用できると期待される。(2)発生早期におけるVEGF受容体の役割をキナーゼ阻害剤などにより解析し、受容体発現細胞(ヘマンジオブラスト)をVEGF-A濃度勾配に依存して頭部へ遊走させるのが重要な機能であることを明らかにした。(3)FRS2ファミリードッキング分子FRS2αの変異マウスの解析より、FRS2αが、FGF受容体によって活性化することにより、胎盤及び網膜の幹細胞の維持、増殖に重要な役割を果たすことが明らかとなった。(4)FRS2βは、EGF受容体シグナル伝達を抑制的に制御することが示された。(5)上皮特異的な増殖抑制・分化誘導作用を有するPKCηは、cyclin D1の発現抑制とトランスグルタミナーゼ1の酵素活性上昇を伴って、E-cadherin、β/γ-cateninを発現するヒト食道扁平上皮癌細胞の増殖を抑制した。この増殖抑制能は、ヌードマウスを用いた皮膚担癌実験により、個体レベルでも確認された。

Research Products

• [Journal Article] Membrane-fixation of VEGFR1 ligand-binding domain is important for vasculogenesis/angiogenesis in mice. (2005)
  • Author(s): Hiratsuka, S., Nakao, K., Nakamura, K., Katsuki, M., Maru, Y., Shibuya, M.
  • Journal Title: Mol.Cell.Biol. 25 Pages: 346-354
• [Journal Article] Essential role of Flk-1 (vascular endothelial growth factor receptor-2) tyrosine residue-1173 in vasculogenesis in mice. (2005)
  • Author(s): Sakurai, Y., Ohgimoto, K., Kataoka, Y., Yoshida, N., Shibuya, M.
  • Journal Title: Proc.Natl.Acad.Sci.USA 102 Pages: 1076-1081
• [Journal Article] A Novel Snake Venom Vascular Endothelial Growth Factor (VEGF) Predominantly Induces Vascular Permeability through Preferential Signaling via VEGF Receptor-1. (2004)
  • Author(s): Takahashi, H., Hattori, S., Iwamatsu, A., Takizawa, H., Shibuya, M.
  • Journal Title: J.Biol.Chem. 279 Pages: 46304-46314
• [Journal Article] Tyrosine phosphorylation sites on FRS2alpha responsible for Shp2 recruitment are critical for induction of lens and retina. (2004)
  • Author(s): Gotoh, N., Ito, M., Yamamoto, S., Yoshino, I, Song, N., Wang, Y., Lax, I., Schlessinger, J., Shibuya, M., Lang, R.A.
  • Journal Title: Proc.Natl.Acad.Sci.USA 101 Pages: 17144-17149
• [Journal Article] Development of primitive and definitive hematopoiesis from nonhuman primate embryonic stem cells in vitro. (2004)
  • Author(s): Umeda, K., Heike, T., Yoshimoto, M., Shiota, M., Suemori, H., Luo, H.Y., Chui, D.H., Torii, R., Shibuya, M., Nakatsuji, N., Nakahata, T.
  • Journal Title: Development 131 Pages: 1869-1879
• [Journal Article] The c-Cb1/CD2AP complex regulates VEGF-induced endocytosis and degradation of Flt-1 (VEGFR-1). (2004)
  • Author(s): Kobayashi, S., Sawano, A., Nojima, Y., Shibuya, M., Maru, Y.
  • Journal Title: FASEB J. 18 Pages: 929-931
• [Publications] Kiba, A., Yabana, N., Shibuya, M.: "A set of loop-1 and -3 structures in the novel VEGF family member, VEGF-ENZ-7, is essential for the activation of VEGFR-2 signaling"J.Biol.Chem.. 278. 13453-13461 (2003)
• [Publications] Kiyono, M., Shibuya, M.: "Bone morphogenetic protein 4 mediates apoptosis of capillary endothelial cells during rat pupillary membrane regression"Mol.Cell Biol.. 23. 4627-4636 (2003)
• [Publications] Autiero, M., Waltenberger, J., Communi, D., Kranz, A., Moons, L., Lambrechts, D., Kroll, J., Plainsance, S., De Mol, M., Bono, F., Kliche, S., Fellbrich, G., Ballmer-Hofer, K., Maglione, D., Mayr-Beyrle, U., Dewerchin, M., Dombrowski, S., Stanimovic, D., Humnelen, P.V., Dehio, C., Hicklin, D., Persico, G., Herbert, J.M., Communi, D., Shibuya, M., Collen, D., Conway, E.M., Carmeliet, P.: "Role of P1GF in the intra-and inter-molecular crolltalk between VEGF receptors Flt1 and Flk1"Nature Med.. 9. 936-943 (2003)
• [Publications] Sasaki, A., Taketomi, T., Kato, R., Saeki, K., Nonami, A., Sasaki, M., Kuriyama, M., Saito, N., Shibuya, M., Yoshimura: "Mammalian Sprouty4 suppresses Ras-independent ERK activation by binding to Raf1"Nature Cell Biol.. 5. 427-432 (2003)
• [Publications] Gotoh, N., Laks, S., Nakashima, M., Lax, I., Schlessinger, J.: "FRS2 family docking proteins with overlapping roles in activation of MAP kinase have distinct spatial-temporal patterns of expression of their transcripts"FEBS lett.. (in press).
• [Publications] Matsumura, M, Tanaka, N., Kuroki, T., Ichihashi M, Ohba, M.: "The η isoform of protein kinase C inhibits UV-induced activation of caspase-3 in normal human keratinocytes"Biochem.Biophys.Res.Commun.. 303. 350-356 (2003)
• [Publications] Shibuya, M.: "Vascular endothelial growth factor receptor family genes : When did the three genes phylogenetically segregate?"Biol. Chem.. 383. 1573-1579 (2002)
• [Publications] Yabana, N., Shibuya, M.: "Adaptor Protein APS Binds the NH_2-Terminal Autoinhihitory Domain of Guanine Nucleotide Exchange Factor Vav3 and Augments its Activity"Oncogene. 21. 7720-7729 (2002)
• [Publications] Hiratsuka, S., Nakamura, K., Iwai, S., Murakami, M., Itoh, T., Kijima, H., Shipley, J.M., Senior, R.M., Shibuya, M.: "MIMP9 induction by Vascular Endothelial Growth Factor Receptor-1 is involved in lung specific metastasis"Cancer Cell. 2. 289-300 (2002)
• [Publications] Kiba, A., Yabana, N., Shibuya, M.: "A set of loop-1 and -3 structures in the novel VIEGF family member, VEGF-ENZ-7, is essential for the activation of VEGFR-2 signaling"J. Biol. Chem.. (in press). (2003)
• [Publications] Nakamura, T., Komiya, M., Sone, K., Hirose, E., Gotoh, N., Morii, H.Ohta, Y., Mori, N.: "Grit, a GTPase-activating protein for the Rho family, regulates neurite extension through association with the TrkA receptor and N-Shc and Crk/Crk adapter molecules"Mol. Cell. Biol.. 22. 8721-8734 (2002)
• [Publications] Matsumura M, Tanaka N., Kuroki T., Ichihashi M, Ohba M.: "The η isoform of protein kinase C inhibits UV-induced activation of caspase-3 in normal human keratinocytes"Biochem. Biophys. Res. Commun.. (in press). (2003)
• [Publications] Sawano, A., Iwai, S., Sakurai, Y., Ito, M., Shitara, K., Nakahata, T., Shibuya, M.: "Vascular Endothelial Growth Factor Receptor-1(Flt-1) is a Novel Cell Surface Marker for the Lineage of Monocyte-Macrophages in humans"Blood. 97. 785-791 (2001)
• [Publications] Hiratsuka, S., Maru, Y., Okada, A., Seiki, M., Noda, T., Shibuya, M.: "Involvement of Fit-1 Tyrosine Kinase (Vascular Endothelial Growth Factor Receptor-1) in Pathological Angiogenesis"Cancer Res.. 61. 1207-1213 (2001)
• [Publications] Luo, J.-C., Shibuya, M.: "A variant of nuclear localization signal of bipartite-type is required for the nuclear translocation of hypoxia inducible factors (1α,2α and 3α)"Oncogene. 20. 1435-1444 (2001)
• [Publications] Kin, Y., Shibuya, M., Maru, Y.: "Inhibition of protein kinase C delta has negative effect on anchorage-independent growth of BCR-ABL-transformed Rat1 cells"Leuk. Res.. 25. 821-825 (2001)
• [Publications] Takahashi, T., Yamaguchi, S., Chiba, K., Shibuya, M.: "A single autophosphorylation site on KDR/Flk-1 is essential for VEGF-A-dependent activation of PLC-γ and DNA synthesis in vascular endothelial cells"EMBO J.. 20. 2768-2778 (2001)
• [Publications] Maru, Y., Bergmann, E., Coin, F., Egly, J., Shibuya, M.: "TFIIH functions are altered by the P21OBCR-ABL oncoprotein produced on the Philadelphia chromosome"Mutat. Res.. 483. 83-88 (2001)
• [Publications] Maru,Y.,Hirosawa,H.,and Shibuya,M: "An oncogenic form of the Flt-1 kinase has a tubulogenic potential in a sinusoidal endothelial cell line."Eur.J.Cell.Biol.. 79. 130-143 (2000)
• [Publications] Shibuya,M.: "Structure and dual function of vascular endothelial growth factor receptor-1 (Flt-1)."Int.J.Biochem.Cell Biol.,. (in press.). 2001
• [Publications] Kanno S,Oda N,Abe M,Terai Y,Ito M,Shitara K,Tabayashi K,Shibuya M,Sato Y.: "Roles of two VEGF receptors, Flt-1 and KDR, in the signal transduction of VEGF effects in human vascular endothelial cells."Oncogene. 19. 2138-2146 (2000)
• [Publications] Shibuya,M.: "Development of the liver vascular system, in Morphogenesis of Endothelium, edited by W.Risau, and G.M.Rubanyi,"Harwood Academic Publishers,. 175-188 (2000)
• [Publications] Sawano,A.,Iwai,S.,Sakurai,Y.,Ito,M.,Shitara,K,Nakahata,T.,and Shibuya,M.: "Vascular Endothelial Growth Factor Receptor-1 (Flt-1) is a Novel Cell Surface Marker for the Lineage of Monocyte-Macrophages in Humans."Blood.. 97. 785-791 (2001)
• [Publications] Chen H,Ikeda U,Shimpo M,Maeda Y,Shibuya M,Ozawa K,Shimada K: "Inhibition of vascular endothelial growth factor activity by tranfection with the soluble FLT-1 gene."J Cardiovasc Pharmacol. 36. 498-502 (2000)
• [Publications] Maeda Y,Ikeda U,Shimpo M,Shibuya M,Monahan J,Urabe M,Ozawa K,Shimada K.: "Adeno-associated virus-mediated vascular endothelial growth factor gene transfer into cardiac myocytes."J Cardiovasc Pharmacol.. 4. 438-443 (2000)
• [Publications] Takahashi,T.and Shibuya,M.: "The overexpression of PKCδ is involved in vascular endothelial growth factor-resistant apoptosis in cultured primary sinusoidal endothelial cells."Biochem Biophys Res Commun.. 280. 415-420 (2001)
• [Publications] Hiratsuka,S.,Maru,Y.,Okada,A.,Seiki,M.,Noda,T.,and Shibuya,M.: "Involvement of Flt-1 Tyrosine Kinase (Vascular Endothelial Growth Factor Receptor-1) in Pathological Angiogenesis"Cancer Res.,. (in press). (2001)
• [Publications] Luo,J.-C.,and Shibuya,M.: "A variant of nuclear localization signal of bipartite-type is required for the nuclear translocation of hypoxia inducible factors (1α, 2α and 3α)."Oncogene,. (in press). (2001)