血液・血管系幹細胞の未分化性維持機構の解析

• Project/Area Number: 14081204
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kumamoto University
• Principal Investigator: 小川 峰太郎 熊本大学, 発生医学研究センター, 教授 (70194454)
• Co-Investigator(Kenkyū-buntansha): 坂本 比呂志 熊本大学, 発生医学研究センター, 助手 (00347014)
• Project Period (FY): 2002 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥48,700,000 (Direct Cost: ¥48,700,000); Fiscal Year 2006: ¥8,500,000 (Direct Cost: ¥8,500,000); Fiscal Year 2005: ¥7,000,000 (Direct Cost: ¥7,000,000); Fiscal Year 2004: ¥11,600,000 (Direct Cost: ¥11,600,000); Fiscal Year 2003: ¥10,800,000 (Direct Cost: ¥10,800,000); Fiscal Year 2002: ¥10,800,000 (Direct Cost: ¥10,800,000)
• Keywords: 造血肝細胞 / 血管内皮細胞 / 胚性肝細胞 / 転写因子 / 細胞系譜 / 細胞接着 / 血管新生 / 細胞分化 / 造血幹細胞 / 胚性幹細胞 / リンパ球 / 多能性 / 個体発生 / 中胚葉 / 血液細胞 / 心筋細胞 / 試験管内分化

Research Abstract

1.血球分化能を持つ血管内皮細胞、中胚葉及び血管内皮から発生した造血前駆細胞をそれぞれ特異的なマーカーを利用して同定できることを明らかにした。
2.マウス胚体内組織AGM領域の血管内皮は造血幹細胞の発生起源の一つとされているが、AGM領域の血管内皮細胞はリンパ球産生に強くバイアスがかかった血液分化能しか持たないことを明らかにした。
3.転写因子c-Mybを誘導的に強制発現するES細胞を作製し、血管内皮細胞が持つ血液細胞発生プログラムにc-Mybが関与し得ることを、試験管内分化実験系により明らかにした。
4.転写因子c-Mybを欠損するES細胞に外来のc-Myb遺伝子を誘導的に発現させる実験系を構築し、血液細胞の分化過程においてc-Mybの発現が量的にも時間的にも厳密に制御されなければならないことを明らかにした。
5.ES細胞から分化誘導した血管内皮細胞において細胞接着装置の動態を解析したところ、血管内皮細胞は細胞同士の接着を維持しながら高い運動1生を保持し、細胞接着装置が動的にリモデリングされることを明らかにした。
6.転写因子Foxo1の生理的な機能に関して遺伝子を標的破壊したES細胞及びマウスを用いて解析し、血管内皮細胞がVEGFに対して適切に応答し正常な形態変化と血管新生を行うためにFoxo1が必要であることを明らかにした。
7.心筋梗塞後に、新生血管に動員される細胞及び骨髄中の血液細胞の一部において、血管内皮に特異的なVE-カドヘリン遺伝子プロモーターが活性化することを、トランスジェニックマウスを用いた解析より明らかにした。
8.Presenilin-1遺伝子を欠損するES細胞から分化誘導した血管内皮細胞の解析により、Presenilin-1が血管内皮細胞の増殖をそのβカテニン結合領域を介して負に制御していることを明らかにした。

Research Products

• [Journal Article] B cell potential can be obtained from pre-circulatory yolk sac, but with low frequency. (2007)
  • Author(s): 杉山 大介
  • Journal Title: Dev. Biol. 301 Pages: 53-61
• [Journal Article] Endothelial cell motility is compatible with functional integrity. (2007)
  • Author(s): 郭 仁勇
  • Journal Title: J. Cell. Physiol. 211 Pages: 327-335
• [Journal Article] Characterization of GATA-1^+ hemangioblastic cells in the mouse embryo. (2007)
  • Author(s): 横溝 智雅
  • Journal Title: EMBO J. 26 Pages: 184-196
• [Journal Article] Presenilin-l controls the growth and differentiation of endothelial progenitor cells through its β-catenin-binding region. (2006)
  • Author(s): 中島 光業
  • Journal Title: Cell Biol. Int. 30 Pages: 239-243
• [Journal Article] Cardiac ischemia activates vascular endothelial cadherin promoter in both pre-existing vascular cells and bone marrow cells involved in neovascularization. (2006)
  • Author(s): 小形 直子
  • Journal Title: Circ. Res. 98 Pages: 897-904
• [Journal Article] Proper levels of c-Myb are discretely defined at distinct steps of hematopoietic cell development. (2006)
  • Author(s): 坂本 比呂志
  • Journal Title: Blood 108 Pages: 896-903
• [Journal Article] Over-expression of c-Myb increases the frequency of hemogenic precursors in the endothelial cell population. (2006)
  • Author(s): 戴 国友
  • Journal Title: Genes Cells 11 Pages: 859-870
• [Journal Article] Accelerated acquisition of permeability barrier function in the skin of presenilin-1-deficient embryos. (2006)
  • Author(s): 中島 光業
  • Journal Title: Arch. Dermatol. Res. 298 Pages: 339-345
• [Journal Article] High level of endothelial cell-specific gene expression by a combination of the 5' flanking region and the 5'half of the first intron of the VE-cadherin gene. (2005)
  • Author(s): 久恒 洋
  • Journal Title: Blood 105・12 Pages: 4657-4663
• [Journal Article] Cell adhesion to ephrinb2 is induced by EphB4 independently of its kinase activity. (2004)
  • Author(s): 坂本 比呂志
  • Journal Title: Biochem.Biophys, Res. Comm. 321 Pages: 681-687
• [Journal Article] Cell adhesion to ephrinb2 is induced by EphB4 independently of its kinase activity. (2004)
  • Author(s): 坂本比呂志
  • Journal Title: Biochem.Biophys.Res.Comm. 321 Pages: 681-687
• [Journal Article] Hemogenic and non-hemogenic endothelium can be distinguished by the activity of fetal liver kinase (FlK)-l promoter/enhancer during mouse embryogenesis. (2003)
  • Author(s): 平位 秀世
  • Journal Title: Blood 101・3 Pages: 886-893
• [Journal Article] Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells. (2003)
  • Author(s): 日高 京子
  • Journal Title: FASEB J. 17 Pages: 740-742
• [Journal Article] Erythropoiesis from acetyl LDL incorporating endothelial cells at the pre-liver stage. (2003)
  • Author(s): 杉山 大介
  • Journal Title: Blood 101・12 Pages: 4733-4738
• [Journal Article] Putative intermediate precursor between hematogenic endothelial cells and blood cells in the developing embryo. (2003)
  • Author(s): Fraser Stuart
  • Journal Title: Develop. Growth Differ. 45 Pages: 63-75
• [Journal Article] Recombination signal sequence-binding protein Jk alters mesoderm cell fate decisions by suppressing cardiomyogenesis. (2003)
  • Author(s): Schroeder Timm
  • Journal Title: Proc. Natl. Acad. Sci. U.S.A. 100・7 Pages: 4018-4023
• [Journal Article] Hemogenic and non-hemogenic endothelium can be distinguished by the activity of fetal liver kinase (Flk)-1 promoter/enhancer during mouse embryogenesis. (2003)
  • Author(s): 平位秀世
  • Journal Title: Blood 101・3 Pages: 886-893
• [Journal Article] Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells. (2003)
  • Author(s): 日高京子
  • Journal Title: FASEB J. 17 Pages: 740-742
• [Journal Article] Erythropoiesis from acetyl LDL incorporating endothelial cells at the pre-liver stage. (2003)
  • Author(s): 杉山大介
  • Journal Title: Blood 101・12 Pages: 4733-4738
• [Journal Article] Putative intermediate precursor between hematogenic endothelial cells and blood cells in the developing embryo. (2003)
  • Author(s): Fraser Stuart
  • Journal Title: Develop.Growth Differ. 45 Pages: 63-75
• [Journal Article] Recombination signal sequence-binding protein JK alters mesoderm cell fate decisions by suppressing cardiomyogenesis. (2003)
  • Author(s): Schroeder Timm
  • Journal Title: Proc.Natl.Acad.Sci.U.S.A. 100・7 Pages: 4018-4023
• [Journal Article] Embryonic stem cell differentiation as a model to study hematopoietic and endothelial cell development. (2002)
  • Author(s): Fraser Stuart
  • Journal Title: Methods Mol.Biol. 185 Pages: 71-81
• [Journal Article] Definitive hematopoietic commitment within the embryonic vascular endothelial-cadherin+ population. (2002)
  • Author(s): Fraser Stuart
  • Journal Title: Exp.Hematol. 30・9 Pages: 1070-1078
• [Journal Article] SCL/tal-1 dependent process determines a competence to select the definitive hematopoietic lineage prior to endothelial differentiation. (2002)
  • Author(s): 遠藤 充浩
  • Journal Title: EMBO J. 21・24 Pages: 6700-6708
• [Journal Article] SCL/tal-1 dependent process determines a competence to select the definitive hematopoietic lineage prior to endothelial differentiation. (2002)
  • Author(s): 遠藤充浩
  • Journal Title: EMBO J. 21・24 Pages: 6700-6708
• [Journal Article] Distinct hemogenic potentials of endothelial cells and CD41^+ cells in the mouse embryos.
  • Author(s): 橋本 和明
  • Journal Title: Dev. Growth Differ. (印刷中)
• [Journal Article] Presenilin-1 controls the growth and differentiation of endothelial progenitor cells through its β-catenin-binding region.
  • Author(s): 中島 光業
  • Journal Title: Cell Biol.Int. 印刷中
• [Journal Article] Cardiac ischemia activates vascular endothelial cadherin promoter in both pre-existing vascular cells and bone marrow cells involved in neovascularization.
  • Author(s): 小形 直子
  • Journal Title: Circ.Res, 印刷中
• [Publications] Fraser Stuart: "Embryonic stem cell differentiation as a model to study hematopoietic and endothelial cell development."Methods Mol.Biol.. 185. 71-81 (2002)
• [Publications] Fraser Stuart: "Definitive hematopoietic commitment within the embryonic vascular endothelial- cadherin+ population"Exp.Hematol.. 30・9. 1070-1078 (2002)
• [Publications] 遠藤充浩: "SCL/tal-1 dependent process determines a competence to select the definitive hematopoietic lineage prior to endothelial differentiation."EMBO J.. 21・24. 6700-6708 (2002)
• [Publications] 平位秀世: "Hemogenic and non-hemogenic endothelium can be distinguished by the activity of fetal liver kinase (Flk)-1 promoter/enhancer during mouse embryogenesis."BLOOD. 101・3. 886-893 (2003)
• [Publications] 日高京子: "Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells."FASEB J.. 17. 740-742 (2003)
• [Publications] 杉山大介: "Erythropoiesis from acetyl LDL incorporating endothelial cells at the pre-liver stage."BLOOD. 101・12. 4733-4738 (2003)
• [Publications] Fraser Stuart: "Putative intermediate precursor between hematogenic endothelial cells and blood cells in the developing embryo"Develop.Growth Differ.. 45. 63-75 (2003)
• [Publications] Schroeder Timm: "Recombination signal sequence-binding protein Jk alters mesodermal cell fate decisions by suppressing cardiomyogenesis."Proc.Natl.Acad.Sci.U.S.A.. 100・7. 4018-4023 (2003)
• [Publications] Fraser Stuart: "Embryonic stem cell differentiation as a model to study hematopoietic and endothelial cell development"Methods in Molecular Biology. 185. 71-78 (2002)
• [Publications] Fraser Stuart: "Definitive hematopoietic commitment within the embryonic vascular endothelial-cadherin + population"Experimental Hematology. 30・9. 1070-1078 (2002)
• [Publications] 遠藤充浩: "SCL/tal-1 dependent process determines a competence to select the definitive hematopoietic lineage prior to endothelial differentiation"The EMBO Journal. 21・24. 6700-6708 (2002)
• [Publications] 平位秀世: "Hemogenic and non-hemogenic endothelium can be distinguished by the activity of fetal liver kinase (Flk)-1 promoter/enhancer during mouse"BLOOD. 101・3. 886-893 (2003)
• [Publications] 日高京子: "Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells"The FASEB Journal. (印刷中).
• [Publications] 杉山大介: "Erythropoiesis from acety1 LDL incorporating endothelial cells at the preliver stage"BLOOD. (印刷中).