2000 年度実績報告書

血管構築の分子機構

研究課題

研究課題/領域番号	09281101
研究機関	東北大学
研究代表者	佐藤靖史東北大学, 加齢医学研究所, 教授 (50178779)
研究分担者	三井洋司生命工学工業技術研究所, 首席研究官西川伸一京都大学, 大学院・医学研究科, 教授 (60127115) 渋谷正史東京大学, 医科学研究所, 教授 (10107427) 藤原敬己国立循環器病センター研究所, 循環器形態部長 (10190092) 月田承一郎京都大学, 大学院・医学研究科, 教授 (50155347)
キーワード	Ets-1 / アポトーシス / VEGF / シグナル伝達 / ES細胞 / 分化 / claudin-5 / テロメラーゼ
研究概要	(1)転写因子Ets-1は血管新生を促進するが、今回このEts-1が血管内皮細胞のアポトーシスを促進すること、さらにその機序としてBid,cytochrome p450,caspase-4,p27,p21などpro-apoptoticな遺伝子の発現を2倍以上に増強し、DAD-1,AXL,Cox-2,IAP-2,MDM-2などanti-apoptoticな遺伝子の発現を1/2以下に減弱する。(2)VEGF-Aによる細胞内シグナル伝達に関して、ERKとp38の活性化の経路が大血管の内皮細胞と細小血管の内皮細胞では異なっており血管新生に関与する細小血管内皮細胞ではRasを介したシグナル伝達が主体である。(3)トランスジェニックマウスの解析からVEGF-EはVEGF-Aと同様に血管新生を誘導するが、血管透過性促進作用は認められない。(4)VEGF2型受容体(KDR)の自己リン酸化部位の同定は1175Yとl214Yであり、特にPLCgのC末側のSH2ドメインがリン酸化1175Yと結合し、MAPKの活性化とDNA合成の誘導に必須である。(5)MadCAM-1が発生初期の静脈に特異的に発現し、MadCAM-1陽性静脈内皮の誘導にBMP-4が促進的効果を有する。(6)GFPキメラ分子をES細胞へ導入し、その細胞を内皮へ分化させて各分化段階の細胞の挙動を様々な条件でモニターすることを可能にした。(7)血管内皮細胞に特異的に発現しているclaudin-5のノックアウトマウスの作製を進め、ES細胞において破壊することに成功し、また、germlineにも通すことができた。(9)ヒト臍帯静脈内皮細胞にhTERTまたはSV40T抗原遺伝子を導入することによって、細胞の分裂寿命が延長し、無限分裂寿命の細胞株を樹立できた。この細胞株は、内皮細胞に作用する新規の増殖因子を産生することが示唆された。

研究成果

(35件)

すべてその他

すべて文献書誌 (35件)

[文献書誌] Tanaka,M., et al.: "Expression of the 37-kDa laminin binding protein in murine lung tumor cell correlates with tumor angiogenesis."Cancer Lett.. 153. 161-168 (2000)
[文献書誌] Kanno,S.,et al.: "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)
[文献書誌] Sato,Y., et al.: "Properties of two VEGF receptors, Flt-1 and KDR, in the signal transduction."Ann.N.Y.Acad.Sci.. 901. 201-207 (2000)
[文献書誌] Nakano,T., et al.: "Angiogenesis Inhibition by Transdominant Mutant Ets-1."J.Cell.Physiol.. 184. 255-262 (2000)
[文献書誌] Sato,Y.: "Molecular mechanism of angiogenesis : transcription factors and their therapeutic relevance."Pharmacol.Ther.. 87. 51-60 (2000)
[文献書誌] Sato,Y., et al.: "Signal transduction and transcriptional regulation of angiogenesis."Adv.Exp.Med.Biol.. 476. 109-115 (2000)
[文献書誌] Sato,Y.: "Role of ETS family transcription factors in vascular development and angiogenesis."Cell Struct.Funct.. (in press). (2001)
[文献書誌] Teruyama,K., et al.: "Role of transcription factor Ets-1 in the apoptosis of human vascular endothelial cells."J.Cell.Physiol.. (in press). (2001)
[文献書誌] Yashima,R., et al.: "Heterogeneity of the signal transduction pathways for VEGF-indued MAPKs activation in human vascular endothelial cells."J.Cell.Physiol.. (in press). (2001)
[文献書誌] Maru,Y., et al.: "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)
[文献書誌] Chen,H., et al.: "Inhibition of vascular endothelial growth factor activity by transfection with the soluble FLT-1 gene."J.Cardiovasc.Pharmacol.. 36. 498-502 (2000)
[文献書誌] Maeda,Y., et al.: "Adeno-associated virus-mediated vascular endothelial growth factor gene transfer into cardiac myocytes."J.Cardiovasc.Pharmacol.. 36. 438-443 (2000)
[文献書誌] Sawano,A., et al.: "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)
[文献書誌] Takahashi,T., et al.: "The overexpression of PKCd is involved in vascular endothelial growth factor-resistant apoptosis in cultured primary sinusoidal endothelial cells."Biochem.Biophys.Res.Commun.. 280. 415-420 (2001)
[文献書誌] Hiratsuka,S., et al.: "Involvement of Flt-1 Tyrosine Kinase (Vascular Endothelial Growth Factor Receptor-l ) in Pathological Angiogenesis."Cancer Res.. (in press). (2001)
[文献書誌] Luo,J.-C., et al.: "A variant of nuclear localization signal of bipartite-type is required for the nuclear translocation of hypoxia inducible factors (la,2a and 3a)."Oncogene. (in press). (2001)
[文献書誌] Shibuya,M.: "Structure and dual function of vascular endothelial growth factor receptor-1 (Flt-1)."Int.J.Biochem.Cell Biol.. (in press). (2001)
[文献書誌] Tsukita,S.h., et al.: "Pores in the wall: Claudins constitute tight junction strands containing aqueous pores."J.Cell Biol.. 149. 13-16 (2000)
[文献書誌] Fujita,K., et al.: "Clostridium perfringens enterotoxin binds to the second extracellular loop of claudin-3,a tight junction integral membrane protein."FEBS Lett.. 476. 258-261 (2000)
[文献書誌] Sitou,M., et al.: "Complex phenotype of mice lacking occludin, a component of tight junction strands."Mol.Cell Biol.. 11. 4131-4142 (2000)
[文献書誌] Tsukita,Sh.: "Tight junctions involving occludin and claudins."Cornea. 19. S142-145 (2000)
[文献書誌] Tsukita,S.h., et al.: "The structure and function or claudins, cell adhesion molecules at tight junctions."Ann.N.Y.Acad.Sci.. 915. 129-135 (2000)
[文献書誌] Furuse,M., et al.: "Conversion of Zonula Occludens from tight to leaky strand type by introducing claudin-2 into MDCK I cells."J.Cell Biol.. (in press). (2001)
[文献書誌] Tsukita,S.h., et al.: "Multifunctional strands of tight junctions."Nature Review (Mol.Cell Biol.). (in press). (2001)
[文献書誌] Tanaka,Y., et al.: "Endothelin-1 is involved in the growth promotion of vascular smooth muscle cells by hyaluronic acid."Int.J.Cardiol.. 76. 39-47 (2000)
[文献書誌] Kaul,S.C., et al.: "Transcriptional inactivation of p53 by deletions and single amino acid changes in mouse mot-1 protein."Biochem.Biophys.Res.Commun.. 279. 602-606 (2000)
[文献書誌] Nishikawa,S.I., et al.: "All B cells are progeny of endothelial cells : a new perspective."Immunol Rev.. 175. 112-119 (2000)
[文献書誌] Kubo,H., et al.: "Vascular Endothelial Growth Factor Receptor-3 is essential for maintenance of integrity of endothelial cell lining during tumor angiogenesis. "Blood. 96. 546-553 (2000)
[文献書誌] Yamashita,J., et al.: "Embryonic stem cell derived Flk1-positive cells serve as vascular progenitors."Nature. 408. 92-96 (2000)
[文献書誌] Nagaoka,H., et al.: "Ras mediated effector patheways responsible for pre-B cell survival, which is essential for the developmental progression to the late pre-B cell stage."J.Exp.Med.. 192. 171-181 (2000)
[文献書誌] Yokomizo,T., et al.: "Requirement of Runx1/AML1/PEBP2B for the generation of hematopoietic and endothelial cells."Genes to Cells. 6. 13-24 (2001)
[文献書誌] Hemmi,H., et al.: "Temporal and spatial localization of osteoclasts in colonies from embryonic stem cells."Biochem.Biophys.Res.Commun.. 280. 526-534 (2001)
[文献書誌] Stacker,S.A., et al.: "Vascular endothelial growth factor-D promotes tumor growth and spread via the lymphatics. "Nature Med.. 7. 186-191 (2001)
[文献書誌] Makinen,T., et al.: "Inhibition of lymphangiogenesis and lymphedema in transgenic mice expressing soluble VEGF receptor 3."Nature Med. 7. 199-205 (2001)
[文献書誌] 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)

2000 年度 実績報告書

血管構築の分子機構

研究代表者

佐藤 靖史 東北大学, 加齢医学研究所, 教授 (50178779)

研究成果

[文献書誌] Tanaka,M., et al.: "Expression of the 37-kDa laminin binding protein in murine lung tumor cell correlates with tumor angiogenesis."Cancer Lett.. 153. 161-168 (2000)

[文献書誌] Kanno,S.,et al.: "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)

[文献書誌] Sato,Y., et al.: "Properties of two VEGF receptors, Flt-1 and KDR, in the signal transduction."Ann.N.Y.Acad.Sci.. 901. 201-207 (2000)

[文献書誌] Nakano,T., et al.: "Angiogenesis Inhibition by Transdominant Mutant Ets-1."J.Cell.Physiol.. 184. 255-262 (2000)

[文献書誌] Sato,Y.: "Molecular mechanism of angiogenesis : transcription factors and their therapeutic relevance."Pharmacol.Ther.. 87. 51-60 (2000)

[文献書誌] Sato,Y., et al.: "Signal transduction and transcriptional regulation of angiogenesis."Adv.Exp.Med.Biol.. 476. 109-115 (2000)

[文献書誌] Sato,Y.: "Role of ETS family transcription factors in vascular development and angiogenesis."Cell Struct.Funct.. (in press). (2001)

[文献書誌] Teruyama,K., et al.: "Role of transcription factor Ets-1 in the apoptosis of human vascular endothelial cells."J.Cell.Physiol.. (in press). (2001)

[文献書誌] Yashima,R., et al.: "Heterogeneity of the signal transduction pathways for VEGF-indued MAPKs activation in human vascular endothelial cells."J.Cell.Physiol.. (in press). (2001)

[文献書誌] Maru,Y., et al.: "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)

[文献書誌] Chen,H., et al.: "Inhibition of vascular endothelial growth factor activity by transfection with the soluble FLT-1 gene."J.Cardiovasc.Pharmacol.. 36. 498-502 (2000)

[文献書誌] Maeda,Y., et al.: "Adeno-associated virus-mediated vascular endothelial growth factor gene transfer into cardiac myocytes."J.Cardiovasc.Pharmacol.. 36. 438-443 (2000)

[文献書誌] Sawano,A., et al.: "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)

[文献書誌] Takahashi,T., et al.: "The overexpression of PKCd is involved in vascular endothelial growth factor-resistant apoptosis in cultured primary sinusoidal endothelial cells."Biochem.Biophys.Res.Commun.. 280. 415-420 (2001)

[文献書誌] Hiratsuka,S., et al.: "Involvement of Flt-1 Tyrosine Kinase (Vascular Endothelial Growth Factor Receptor-l ) in Pathological Angiogenesis."Cancer Res.. (in press). (2001)

[文献書誌] Luo,J.-C., et al.: "A variant of nuclear localization signal of bipartite-type is required for the nuclear translocation of hypoxia inducible factors (la,2a and 3a)."Oncogene. (in press). (2001)

[文献書誌] Shibuya,M.: "Structure and dual function of vascular endothelial growth factor receptor-1 (Flt-1)."Int.J.Biochem.Cell Biol.. (in press). (2001)

[文献書誌] Tsukita,S.h., et al.: "Pores in the wall: Claudins constitute tight junction strands containing aqueous pores."J.Cell Biol.. 149. 13-16 (2000)

[文献書誌] Fujita,K., et al.: "Clostridium perfringens enterotoxin binds to the second extracellular loop of claudin-3,a tight junction integral membrane protein."FEBS Lett.. 476. 258-261 (2000)

[文献書誌] Sitou,M., et al.: "Complex phenotype of mice lacking occludin, a component of tight junction strands."Mol.Cell Biol.. 11. 4131-4142 (2000)

[文献書誌] Tsukita,Sh.: "Tight junctions involving occludin and claudins."Cornea. 19. S142-145 (2000)

[文献書誌] Tsukita,S.h., et al.: "The structure and function or claudins, cell adhesion molecules at tight junctions."Ann.N.Y.Acad.Sci.. 915. 129-135 (2000)

[文献書誌] Furuse,M., et al.: "Conversion of Zonula Occludens from tight to leaky strand type by introducing claudin-2 into MDCK I cells."J.Cell Biol.. (in press). (2001)

[文献書誌] Tsukita,S.h., et al.: "Multifunctional strands of tight junctions."Nature Review (Mol.Cell Biol.). (in press). (2001)

[文献書誌] Tanaka,Y., et al.: "Endothelin-1 is involved in the growth promotion of vascular smooth muscle cells by hyaluronic acid."Int.J.Cardiol.. 76. 39-47 (2000)

[文献書誌] Kaul,S.C., et al.: "Transcriptional inactivation of p53 by deletions and single amino acid changes in mouse mot-1 protein."Biochem.Biophys.Res.Commun.. 279. 602-606 (2000)

[文献書誌] Nishikawa,S.I., et al.: "All B cells are progeny of endothelial cells : a new perspective."Immunol Rev.. 175. 112-119 (2000)

[文献書誌] Kubo,H., et al.: "Vascular Endothelial Growth Factor Receptor-3 is essential for maintenance of integrity of endothelial cell lining during tumor angiogenesis. "Blood. 96. 546-553 (2000)

[文献書誌] Yamashita,J., et al.: "Embryonic stem cell derived Flk1-positive cells serve as vascular progenitors."Nature. 408. 92-96 (2000)

[文献書誌] Nagaoka,H., et al.: "Ras mediated effector patheways responsible for pre-B cell survival, which is essential for the developmental progression to the late pre-B cell stage."J.Exp.Med.. 192. 171-181 (2000)

[文献書誌] Yokomizo,T., et al.: "Requirement of Runx1/AML1/PEBP2B for the generation of hematopoietic and endothelial cells."Genes to Cells. 6. 13-24 (2001)

[文献書誌] Hemmi,H., et al.: "Temporal and spatial localization of osteoclasts in colonies from embryonic stem cells."Biochem.Biophys.Res.Commun.. 280. 526-534 (2001)

[文献書誌] Stacker,S.A., et al.: "Vascular endothelial growth factor-D promotes tumor growth and spread via the lymphatics. "Nature Med.. 7. 186-191 (2001)

[文献書誌] Makinen,T., et al.: "Inhibition of lymphangiogenesis and lymphedema in transgenic mice expressing soluble VEGF receptor 3."Nature Med. 7. 199-205 (2001)

[文献書誌] 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)

2000 年度実績報告書

佐藤靖史東北大学, 加齢医学研究所, 教授 (50178779)