Activation of Arterial Wall Cells during Atherogenesis : A study using a in vitor co-culture model system

• Project/Area Number: 15209026
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Circulatory organs internal medicine
• Research Institution: The University of Tokyo
• Principal Investigator: KODAMA Tatsuhiko The University of Tokyo, Research Center for Advanced Science and Technology, Professor, 先端科学技術研究センター・任期付特別教員(教授) (90170266)
• Co-Investigator(Kenkyū-buntansha): NAITO Makoto Niigata University, Graduate School of Medical and Dental Sciences, Professor, 大学院・医歯学総合研究科, 教授 (30045786) ; DOI Takefumi Osaka University, Graduate School of Pharmaceutical Sciences, Professor, 大学院・薬学研究科, 教授 (00211409) ; NOGUCHI Noriko The University of Tokyo, Research Center for Advanced Science and Technology, Associate Professor, 先端科学技術研究センター, 科学技術振興特任教員(特任助教授) (40198578) ; MINAMI Takashi The University of Tokyo, Research Center for Advanced Science and Technology, Associate Professor, 先端科学技術研究センター, 科学技術振興特任教員(特任助教授) (00345141)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥49,270,000 (Direct Cost: ¥37,900,000、Indirect Cost: ¥11,370,000); Fiscal Year 2004: ¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000); Fiscal Year 2003: ¥33,670,000 (Direct Cost: ¥25,900,000、Indirect Cost: ¥7,770,000)
• Keywords: ATHEORSCLEROSIS / ENDOTHELIAL CELL / ADHESION MOLECULE / TRANSCRIPTINAL FACTORS / NFATC / DSCR1 / NRF2 / GATA / Rac1 / コレステロール / ラフト / 炎症 / マイクロアレイ / 血管内皮細胞 / トロンビン / TNF-a / target transgenic mouse / Tie-2

Research Abstract

<Purpose>A new in vitro system for the co-culture of vascular endothelial cell(EC), smooth muscle cell(SMC) and monocyte/macrophage has been developed for the anaylysis of pathogenesis of atherosclerosis.
<Method>We have established a vascular cell under flow. Using this system we examined the molecular pathogenesis of atherosclerosis and endothelial activation.
Results
(1)Oxidative modification of lipoproteins : We established a method for tracing oxidative modification. Transcriptome analysis indicated lyso-PC affects many genes in EC.
(2)Using co-culture system, we found the involvement of NRF2 in shear stress sensing.
(3)Changes in transcriptome were analyzed during inflammation, growth and coagulation in EC. Adding to Ets, GATA and Hex, the role of NFATc in these signal transduction was found. We also found that DSCRl can inhibit NFATc signaling.
(4)We found a novel inhibitor for upregulation of adhesion molecule using co-culture system. This drug is also effective for the treatment of psoriasis.

Research Products

• [Journal Article] The isolation of mouse aortic endothelial cells by a simple method. (2005)
  • Author(s): Kobayashi M, Inoue K, Warabi E, Minami T, Kodama T
  • Journal Title: J Atheroscler Thromb (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression of the LXR{alpha} Protein in Human Atherosclerotic Lesions. (2005)
  • Author(s): Watanabe Y, Jiang S, Takabe W, Ohashi R, Tanaka T, Uchiyama Y, Katsumi K, Iwanari H, Noguchi N, Naito M, Hamakubo T, Kodama T.
  • Journal Title: Arterioscler Thromb Vasc Biol. 25(3) Pages: 622-627
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression of the LXR{alpha} Protein in Human Atherosclerotic Lesions. (2005)
  • Author(s): Watanabe Y, Jiang S, Takabe W, Ohashi R, Tanaka T, Uchiyama Y, Katsumi K, Iwanari H, Noguchi N, Naito M, Hamakubo T, Kodama T.
  • Journal Title: Arterioscler Thromb Vase Biol 25(3) Pages: 622-627
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression of the LXR{alpha} Protein in Human Atherosclerotic Lesions. (2005)
  • Author(s): Watanabe Y., Jiang S, Takabe W, Ohashi R, Tanaka T, Uchiyama Y, Katsumi K, Iwanari H, Noguchi N, Naito M, Hamakubo T, Kodama T.
  • Journal Title: Arterioscler Thromb Vasc Biol. 25(3) Pages: 622-627
• [Journal Article] VEGF- and thrombin-induced termination factor, down syndrome critical region-1,attenuates endothelial cell proliferation, and angiogenesis. (2004)
  • Author(s): Minami T, Horiuchi K, Miura M, Abid R, Takabe W, Kohro T, Ge X, Aburatani H, Hamakubo T, Kodama T, Aird WC.
  • Journal Title: J Biol Chem. 279(48) Pages: 50537-50554
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Effect on endothelial cell gene expression of shear stress, oxygen concentration, and low-density lipoprotein as studied by a novel flow cell culture system. (2004)
  • Author(s): Warabi E, Wada Y, Kajiwara H, Kobayashi M, Koshiba N, Hisada T, Shibata M, Ando J, Tsuchiya M, Kodama T, Noguchi N.
  • Journal Title: Free Radic Biol Med. 1;37(5) Pages: 682-694
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Interaction between Hex and GATA transcription factors in vascular endothelial cells inhibits flk-1/KDR-mediated VEGF signaling. (2004)
  • Author(s): Minami T, Murakami T, Horiuchi K, Miura M, Noguchi T, Miyazaki JI, Hamakubo T, Aird WC, Kodama T.
  • Journal Title: J Biol Chem. 279(20) Pages: 20626-20635
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] VEGF-and thrombin-induced termination factor, down syndrome critical region-1, attenuates endothelial cell proliferation, and angiogenesis. (2004)
  • Author(s): Minami T, Horiuchi K, Miura M, Abid R, Takabe W, Kohro T, Ge X, Aburatani H, Hamakubo T, Kodama T, Aird WC.
  • Journal Title: J Biol Chem. Nov 26;279(48) Pages: 50537-542001
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Effect on endothelial cell gene expression of shear stress, oxygen concentration, and low-density lipoprotein as studied by a novel flow cell culture system (2004)
  • Author(s): Warabi E, Wada Y, Kajiwara H, Kobayashi M, Koshiba N, Hisada T, Shibata M, Ando J, Tsuchiya M, Kodama T, Noguchi N.
  • Journal Title: Free Radic Biol Med. Sep 1;37(5) Pages: 682-694
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] VEGF-and thrombin-induced termination factor, down syndrome critical region-1, attenuates endothelial cell proliferation, and angiogenesis. (2004)
  • Author(s): Minami T, Horiuchi K, Miura M, Abid R, Takabe W, Kohro T, Ge X, Aburatani H, Hamakubo T, Kodama T, Aird WC.
  • Journal Title: J Biol Chem. Sep 23 ([Epubahesd of print])
• [Journal Article] Effect on cell gene expression by shear stress, oxygen concentration, low-density lipoprotein as studied by a novel flow cell culture system. (2004)
  • Author(s): Warabi E, Wada Y, Kajiwara H, Kobayashi M, Koshiba N, Hisada T, Shibata M, Ando J, Tsuchiya M, Kodama T, Noguchi N.
  • Journal Title: Free Radic Biol Med. 1;37(5) Pages: 682-692
• [Journal Article] Interaction between Hex and GATA transcription factors in vascular endothelial cells Inhibits flk-1/KDR-mediated VEGF signaling. (2004)
  • Author(s): Minami T, Murakami T, Horiuchi K, Miura M, Noguchi T, Miyazaki JI, Hamakubo T, Aird WC, Kodama T.
  • Journal Title: J Biol Chem. 279(20) Pages: 20626-20635
• [Journal Article] Thrombin stimulation of VCAM-1 in endothelial cells is mediated by PKC-delta -NF-kappa B and PKC-zeta -GATA signaling pathways. (2003)
  • Author(s): Minami T, Abid MR, Zhang J, King G, Kodama T, _Aird WO.
  • Journal Title: J Biol Chem. 278 Pages: 6976-6984
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Thrombin stimulation of VCAM-1 in endothelial cells is mediated by PKC-delta -NF-kappa B and PKC-zeta -GATA signaling pathways. (2003)
  • Author(s): Minami T, Abid MR, Zhang J, King G, Kodama T, Aird WC.
  • Journal Title: J Biol Chem. 278(9) Pages: 6976-6984
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Thrombin stimulation of VCAM-1 in endothelial cells is mediated by PKC-delta -NF-kappa B and PKC-zeta -GATA signaling pathways. (2003)
  • Author(s): Minami T, Abid MR, Zhang J, King G, Kodama T, Aird WC.
  • Journal Title: J Biol Chem. 278 Pages: 6976-6984
• [Publications] T.Minami, T.Kodama, et al.: "Thrombin and Phenotype Modulation of the Endothelium."Arterioscler.Thromb.Vasc.Biol.. 24. 41-53 (2004)
• [Publications] T.Minami, T.Kodama, et al.: "Ets Motifs are Necessary for Endothelial Cell Specific Expression of a 723-bp Tie-2 Promoter/Enhancer in Hprt-Targeted Transgenic Mice."Arterioscler.Thromb.Vasc.Biol.. 23. 2041-2043 (2003)
• [Publications] N.Noguchi, R.Hanyu, A.Nonaka, Y.Okimoto, T.Kodama: "Inhibition of THP-1 cell adhesion to endothelial cells by α-tocopherol and α-tocotrienol is dependent on intracellular concentration of the antioxidants."Free Rad.Biol.Med. 34. 1614-1620 (2003)
• [Publications] Y.Okimoto, E.Warabi, Y.Wada, E.Niki, T.Kodama, N.Noguchi: "A novel method of following oxidation of low-density lipoprotein using a sensitive fluorescent probe, diphenyl-1-pyrenylphosphine."Free Rad.Biol Med.. 5. 576-585 (2003)