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Elucidation of the mechanism of vascular cell sensing and gene responses to mechanical stresses

Research Project

Project/Area Number 11308035
Research Category

Grant-in-Aid for Scientific Research (A).

Allocation TypeSingle-year Grants
Section一般
Research Field Biomedical engineering/Biological material science
Research InstitutionUniversity of Tokyo

Principal Investigator

ANDO Joji  University of Tokyo, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (20159528)

Co-Investigator(Kenkyū-buntansha) KAMIYA Akira  Nihon University, Interdisciplinary Science Center, Professor, グローバルビジネス研究科, 教授 (50014072)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥27,900,000 (Direct Cost: ¥27,900,000)
Fiscal Year 2000: ¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1999: ¥20,900,000 (Direct Cost: ¥20,900,000)
Keywordsendothelial cells / intracellular signal transduction / calcium signaling / purinoceptor / mechano-sensitive channel / calcium ion channel / adenosine triphosphate / fluid shear stress / 血行力学因子 / 遺伝子 / 転写因子 / チロシンキナーゼ / FAK
Research Abstract

We investigated signal transduction leading to shear stress responsive element-mediated transcription. Bovine endothelial cells were transfected with luciferase reporter vectors containing tandem repeats of TRE, CRE or NFkB element, and exposed to a shear stress of 15 dynes/cm^2. Transcriptional activity via each element began to increase significantly at 3 h after the onset of shear stress and increased further with time, peaking at 12h. These responses to shear stress were significantly suppressed by the treatment of cells with a tyrosine kinase inhibitor (Herbimycin A, 1 mM). Shear stress induced tyrosine phosphorylation of proteins of around 30, 45-65, and 120 kDa including focal adhesion kinase (FAK) in endothelial cells. Overexpression of FRNK (FAK-related non-kinase), which acts as an inhibitor of FAK, markedly inhibited the shear stress-induced increase in transcription via TRE, CRE or NFkB element. These results suggest that tyrosine kinase and FAK signaling are critical for s … More hear stress induction of TRE-, CRE- and NFkB element-mediated transcription in endothelial cells.
We also studied molecular mechanism of Ca^<2+> signaling, including the ion channels responsible for the shear stress-induced Ca^<2+> responses in endothelial cells. Human umbilical vein ECs (HUVECs) loaded with the Ca^<2+> indicator lndo-1/AM were exposed to laminar flow of Hanks' balanced salt solution (HBSS) at various concentrations of ATP and changes in intraceliular Ca^<2+> concentrations ([Ca^<2+>]i) were monitored by confocal laser scanning microscopy. A stepwise increase in flow rate elicited a corresponding stepwise-increase in [Ca^<2+>]i at 250 nmol/L ATP.The flow rate-dependent increase in [Ca^<2+>]i disappeared after the chelation of extracellular Ca^<2+> with EGTA.Antisense oligonucleotides designed to knockout P2X4 purinoceptor expression abolished the flow-induced Ca^<2+> influx. Human embryonic kidney 293 cells showed no Ca^<2+> response to flow at 2μmol/L ATP, but when transfected with P2X4 cDNA they began to show flow-rate-dependent Ca^<2+> influx. P2X4 purinoceptors may have a "shear-transducer" property by which shear stress is perceived directly or indirectly and transmitted into the cell interior via Ca^<2+> signaling. Less

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] R.Korenaga: "Sp1-mediated downregulation of P2X4 receptor gene transcription in endothelial cells exposed to shear stress."Am.J.Physiol Heart Circ.Physiol.. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Kato: "mRNA expression on shape-engineered endothelial cells : Adhesion molecules ICAM-1 and VCAM-1"J.Biomed.Mater Res.. 54. 366-372 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Yamamoto: "Fluid shear stress activates Ca^<2+> influx into human endothelial cells via P2X4 purinoceptors."Circ.Res.. 87. 385-391 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Yamamoto: "P2X4 receptors mediate ATP-induced calcium influx in human vascular endothelial cells."Am.J.Physiol Heart Circ.Physiol.. 279. H285-H292 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Yamaguchi: "The effect of flow on the neutrophil-mediated Ca^<2+> responses In human vascular endothelial cells stimulated by endotoxin."Jpn.J.Surg.. 29. 966-969 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] "Shear Stress-Dependent Regulation of Endothelial Cell Functions In "New Frontier in Vascular Biology ; Thrombosis & Hemostasis"."K.Suzuki et al.eds.,Eibun Press,Osaka. 16 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] J.Ando: "Flow-induced endothelial gene regulation.In"Mechanical Forces and the Endothelium""P.I.Lelkes ed.,Harwood Academic Publishers,London. 15 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Kato, J.Ando, and T.Matsuda: "mRNA expression on shape-engineered endothelial cells : Adhesion molecules ICAM-1 and VCAM-1."J.Biomed.Mater.Res.. 54. 366-372 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Yamamoto, R.Korenaga, A.Kamiya, and J.Ando: "Fluid shear stress activates Ca2+ influx into human endothelial cells via P2X4 purinoceptors."Circ.Res.. 87. 385-391 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Yamamoto, R.Korenaga, A.Kamiya, Z.Qi, M.Sokabe, and J.Ando: "P2X4 receptors mediate ATP-induced calcium influx in human vascular endothelial cells."Am.J.Physiol Heart Circ.Physiol.. 279. H285-H292 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] J.Ando, R.Korenaga, and A.Kamiya: "Shear Stress-Dependent Regulation of Endothelial Cell Functions."In "New Frontier in Vascular Biology ; Thrombosis & Hemostasis", K.Suzuki et al. eds., Eibun Press, Osaka. 199-215 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] J.Ando, R.Korenaga, and A.Kamiya: "Flow-induced endothelial gene regulation."In "Mechanical Forces and the Endothelium", P.I.Lelkes ed., Harwood Academic Publishers, London. 111-126 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Yamaguchi, H.Machida, R.Korenaga, N.Toyama-Sorimachi, J.Ando, M.Miyasaka, T.Matsumoto, H.Nakano, K.Kumada, and M.Takeda: "The effect of flow on the neutrophil-mediated Ca2+ responses in human vascular endothelial cells stimulated by endotoxin."Jpn.J.Surg.. 29. 966-969 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Nagase, J.Abe, K.Takahashi, J.Ando, S.Hirose, and T.Fujita: "Genomic organization and regulation of expression of the lectin-like oxidized low-density lipoprotein receptor (LOX-1) gene."J.Biol.Chem.. 273. 33702-33707 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Nakai, I.Sakuma, T.Ohta, J.Ando, A.Kitabatake, Y.Nakazato, and T.Takahashi: "Permeability characteristics of hemoglobin derivatives across cultured endothelial cell monolayers."J.Lab.Clin.Med.. 132. 313-319 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Murase, N.Kume, R.Korenaga, J.Ando, T.Sawamura, T.Masaki, and T.Kita: "Fluid shear stress transcriptionally induces lectin-like oxidized LDL receptor-1 in vascular endothelial cells."Circ.Res.. 83. 328-333 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Tsuzuki, N.Toyama-Sorimachi, F.Kitamura, H.Tusboi, J.Ando, T.Sakurai, N.Morii, S.Narumiya, and M.Miyasaka: "Intracellular signal-transducing elements involved in transendothelial migration of lymphoma cells."Jpn.J.Cancer Res.. 89. 571-577 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Masatugu, H.Itoh, T-H.Chun, Y.Ogawa, N.Tamura, J.Yamashita, K.Doi, M.Inoue, Y.Fukunaga, N.Sawada, T.Saito, R.Korenaga, J.Ando, and K.Nakao: "Physiologic shear stress suppresses endothelin-converting enzyme-1 expression in vascular endothelial cells."J.Cardiovasc.Pharmacol.. 31. S42-S45 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Isshiki, J.Ando, R.Korenaga, H.Kogo, T.Fujimoto, T.Fujita, and A.Kamiya: "Endothelial Ca2+ waves preferentially originate at specific loci in caveolin-rich cell edges."Proc.Natl.Acad.Sci.U.S.A.. 95. 5009-5014 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Watanabe, R.Takahashi, X-X.Zhang, Y.Goto, H.Hayashi, J.Ando, M.Isshiki, M.Seto, H.Hidaka, I.Niki, and R.Ohno: "An essential role of myosin light-chain kinase in the regulation of agonist- and fluid-flow-stimulated Ca2+ influx in endothelial cells."FASEB J. 12. 341-348 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Kosaki, J.Ando, R.Korenaga, T.Kurokawa, and A.Kamiya: "Fluid shear stress increases the production of granulocyte-macrophage colony-stimulating factor by endothelial cells via mRNA stabilization."Circ.Res.. 81. 794-803 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] R.Korenaga: "Sp1-mediated downregulation of P2X4 receptor gene transcription in endothelial cells exposed to shear stress."Am.J.Physiol Heart Circ.Physiol.. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.Kato: "mRNA expression on shape-engineered endothelial cells : Adhesion molecules ICAM-1 and VCAM-1"J.Biomed.Mater Res.. 54. 366-372 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] K.Yamamoto: "Fluid shear stress activates Ca^<2+> influx into human endothelial cells via P2X4 purinoceptors."Circ.Res.. 87. 385-391 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] K.Yamamoto: "P2X4 receptors mediate ATP-induced calcium influx in human vascular endothelial cells."Am.J.Physiol Heart Circ.Physiol.. 279. H285-H292 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] M.Yamaguchi: "The effect of flow on the neutrophil-mediated Ca^<2+> responses In human vascular endothelial cells stimulated by endotoxin."Jpn.J.Surg.. 29. 966-969 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] J.Ando: "Shear Stress-Dependent Regulation of Endothelial Cell Functions In "New Frontier in Vascular Biology ; Thrombosis & Hemostasis""K.Suzuki et al.eds., Eibun Press, Osaka. 16 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] J.Ando: "Flow-induced endothelial gene regulation.In "Mechanical Forces and the Endothelium""P.I.Lelkes ed., Harwood Academic Publishers, London. 15 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] K. Yamamoto: "P2×4 receptors mediate ATP-induced calcium influx in human vascular endothelial cells"Am. J. Physiol. Heart and Circulatory Physiology. (in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] M. Yamaguchi: "The effect of flow on the neutrophil-mediated Ca^<2+> responses in human vascular endothelial cells stimulated by endotoxin"Jpn. J. Sung.. 29. 966-969 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] J. Ando: "Shear Stress-dependent Regulation of Endothelial Cell Functions. In "New Frontier in Vascular Biology ; Thrombosis & Hemostasis""K. Suzuki et al. eds., Eibun Press, Osaka (in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] J. Ando: "Flow-induced endothelial gene regulation. In "Mechanical Forces and the Endothelium""P. I. Leikes ed., Harwood Academic Publishers, London. 111-126 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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