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2000 Fiscal Year Final Research Report Summary

molecular cloning of self-renewal factor for hematopoietic stem cells and its clinical application

Research Project

Project/Area Number 11357008
Research Category

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

Allocation TypeSingle-year Grants
Section展開研究
Research Field Pediatrics
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

NAKAHATA Tatsutoshi  Kyoto University, Graduate School of Medicine, Professor, 医学研究科, 教授 (20110744)

Co-Investigator(Kenkyū-buntansha) LIN Ying-wei  Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (40283600)
YORIFUJI Tohru  Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (60220779)
KATAMURA Kenji  Kyoto University, Graduate School of Medicine, Lecturer, 医学研究科, 講師 (40185806)
Project Period (FY) 1999 – 2000
Keywordshematopoietic stem cells / self-renewal factor / AGM region / stromal cell line / molecular cloning / mRNA / mouse / novel genes
Research Abstract

Recent studies on the developing mouse embryo have shown that although primitive erythropoiesis is detectable in the yolk sac (YS) at 7.5 days post coitum (dpc), long-term repopulating hematopoietic stem cells (LTR-HSC) are first identified in the aorta-gonad-mesonephros (AGM) region at 10 dpc, prior to such activity being observed in YS and fetal liver, and expand in 11 dpc AGM region, suggesting that the AGM region at 10 to 11 dpc provides a microenvironment suitable for the development of LTR-HSC.These observations prompted us to establish stromal cell lines from the AGM region at 10 to 11 dpc, which would support ex vivo expansion of hematopoietic stem cells. We succeeded the establishment of a novel stromal cell line (AGMA9) from the AGM region of 10.5 dpc mouse embryo. When co-cultured with the AGMA9, lin-Sca-1+c-Kit+ cells isolated form adult mouse bone marrow and human cord blood CD34+ cells, significantly proliferated without additional cytokines. Expanded cocultured human CD34+ cells with AGMA9 for 4 weeks could reconstitute bone marrow of NOD/SCID mice suggesting that the cell line express some novel molecules which affect proliferation of not only murine but also human hematopoietic stem cells. This cell line can now be used to elucidate the molecular mechanisms regulating early hematopoiesis, and provide strategies for manipulation of primitive hematopoietic progenitor/stem cells. In RT-PCR analysis, AGMA9 produced detectable levels of SCF, SDF-1, OSM, IL-6, HGF, MIP-1 γ, Gas6, MCP-3, but no detectable levels of M-CSF EPO, TPO, Flk2/Flt3 ligand, MIP-1 α. We started molecular cloning of a novel self-renewal factor of hematopoietic stem cells using cDNA libraries of AGMA9 and AGMA7 which established from the AGM region at 10.5 dpc, and could not support proliferation of hematopoietic stem cells. We succeeded the cloning of more than ten novel genes and started the analysis of their functions.

  • Research Products

    (64 results)

All Other

All Publications (64 results)

  • [Publications] Futaki M.,Yamashita T.,Yabe M., Kato S.,Nakahata T.: "The IVS4+4A-T mutation of the Fanconi Anemia gene FANCC is not associated with severe phenotype in Japanese patients."Blood. 95. 1493-1498 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sugiyama H.,Nonaka T.,Komoriya K.,Tsuji K.,Nakahata T.: "Peroxisome proliferator-activated receptors negatively regurate mast cell activation"FEBS Letters. 467. 259-262 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ueda Y,Tsuji K,Yoshino H,Ebihara Y,Yagasaki H,Yasukawa K,Nakahata T: "Expansion of human NOD/SCID-repopulating cells by a combination of stem cell factor, Flk2/Flt3 ligand, thrombopoietin and a complex of interleukin-6 and soluble interleukin-6 receptor"J Chin Invest. 105. 1013-1021 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hayashi Y.,Honma Y.,Niitsu N.,Tsuji K.,Nakahata T.: "SN-1, a novel leukemic cell line with t(11;16)(q23;p13): Myeloid characteristics and resistance to retinoids and vitamin D3."Cancer Res.. 60. 1139-1145 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Misawa K.,Nosaka T.,Morita S.,Nakahata T.,Asano S.,Kitamura T.: "A method to identify cDNA based on localization of green fluorescent protein fusion products."Proc.Natl.Acad.Sci.USA. 97. 3062-3066 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ebihara Y,Xu M,Nakahata T,Tsuji K: "Exclusive expression of G-CSF receptor on myeloid progenitors in bone marrow CD34+ cells"Br J Haematol. 109. 153-161 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ueda T,Manabe A,Kikuchi A,Yoshino H,Ebihara Y,Ishii T,Tsuji K,Nakahata T: "Massive pericardial and pleural effusion with anasarca following allogeneic bone marrow transplantation"Int J Hematol. 71. 394-397 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ueda T,Yoshino H,Kobayashi K,Asano S,Nakahata T,Tsuji K: "Hematopoietic repopulating ability of cord blood CD34+ cells in NOD/Shi-scid mice"Stem Cells. 18. 204-213 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsui A.,Sato T.,Maekawa T.,Asano S.,Nakahata T.,Tsuji K.: "Glycoprotein 130 and c-kit signals synergistically induce thrombopoietin production by hematopoietic cells"Int.J.Hematol. 72. 455-462 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sato T,Maekawa T,Watanabe S,Tsuji K,Nakahata T: "Erythroid progenitors differentiate and mature in response to endogenous erythropoietin"J Clin Invest. 106. 263-270 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sasaki S,Ito E,Toki T,Maekawa T,Nakahata T,Igarashi K,Yokoyama M: "Cloning and expression of human B cell-specific transcription factor BACH2 mapped to chromosome 6q15"Oncogene.. 19. 3739-3749 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tanaka Y,Naruse I,Xu M,Nakahata T,Maekawa T,Ishii S: "Extensive brain hemorrhage and embyonic lethality in a mouse null mutant of CREB-binding protein."Mechanisms of Development. 95. 133-145 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishii T,Manabe A,Ebihara Y,Nakahata T,Asano S,Tuji K: "Improvement in bronchiolitis obliterans organizing pneumonia in a chlid after allogeneic bone marrow transplantation by a combination of oral prednisolone and low dose erythromycin"Bone Marrow Transplantation. 26. 907-910 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Muroi J,Yorifuji T,Uematsu A,Kitamura A,Nakahata T: "Molecular and clinical analysis of Japanese patients with 3-hydroxy-3-methylglutaryl CoA lyase(HL) deficiency"Hum Genet.. 107. 320-326 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Muroi J,Yorifuji T,Uematsu A,Nakahata T: "Cerebral infarction and pancreatitis : possible complications of patients with 3-hydroxy-3-methylglutaryl-CoA lyase deficiency"J.Inherit.Metab.Dis.. 23. 636-637 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Higuchi Y,Hattori H,Tsuji M,Asato R,Nakahata T: "Partial seizures in leukoencephalopathy with swelling and a discrepantly mild clinical course"Brain & Development. 22. 387-389 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakahata T.: "Ex vivo expansion of human hematopoietic stem cells"Int J Hematol.. 73(1). 6-13 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoshino H,Ueda T,Kawahata M,Asano S,Nakahata T,Tsuji K: "Natural killer cell depletion by ant asialo GM1 antiserum treatment enhances human hematopoietic stem cell engraftment in NOD/Shi-scid mice"Bone Marrow Transplant. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sugiyama H,Nonaka T,Tsuji K,Nakahata T: "Peroxisome proliferator-activated receptors are expressed in mouse bone marrow-derived mast cells"FEBS Lett,. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ma F,Yang F,Kaneko A,Asano S,Nakahata T,Tsuji K: "Cytokine requirement for the development of T-lineage potential from lymphohematopoietic progenitor cells"Br J Haematol,. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsuoka S,Ebihara Y,Xu M,Ikeda Y,Nakahata T,Tsuji K: "CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Xu M,Matsuoka S,Yang F-C,Asano S,Nakahata T,Tsuji K: "Evidence for the presence of primitive megakaryopoiesis in the early yolk sac"Blood. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamashita T,Nakahata T.: "Current knowledge on the pathophysiology of fanconi anemia : from genes to phenotypes"Int J Hematol. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nishikawa M.,Tahara T.,Nakahata T.,Shimosaka A.: "Role of the microenvironment of the embryonic aorta-gonad-mesonephros region in hematopiesis"Stem Cells. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sawano A.,Iwai S.,Sakurai Y.,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. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsuoka S,Tsuji K,Ikeda Y,Asano S.,Nakahata T: "Generation of definitive hematopoietic stem cells from murine early yolk sac and paraaortic splanchnopleura by AGM region-derived stromal cells"Blood. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ma F.,Wada,M.,Yoshino,H.,Nakahata,T.,Asano,S.,Tsuji,K.: "Development of human lymphohematopoietic stem/progenitor cells defined by CD34 and CD81 expressions"Blood. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 中畑龍俊: "造血幹細胞のex vivo増幅と臨床応用"Biotherapy. 14. 270-275 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 中畑龍俊: "ヒトES細胞と再生医療"BIO Clinica. 15(6). 322-326 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 久保田優,濱畑啓悟,渡邉健一郎,林英蔚,小石誠二,宇佐美郁哉,中畑龍俊,秋山祐一: "小児赤白血病の3症例:予後因子の文献的考察"臨床血液. 41(3). 212-217 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 中畑龍俊: "造血幹細胞と造血前駆細胞"血液・免疫・腫瘍. 5(3). 251-256 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 前川平,石井武文,西原政道,中畑龍俊: "ヒト造血細胞におけるケモカイン受容体CXCR4の発現動態とその臨床的意義."臨床免疫. 33(1). 99-105 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 中畑龍俊: "造血幹細胞のex vivo増幅と臨床応用"遺伝子医学. 4(2). 246-250 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 今井剛,瀧本哲也,秋山祐一,足立壮一,久保田優,中畑龍俊: "寛解導入不能の縦隔原発T細胞性非ホジキンリンパ腫3例の臨床的検討"日小血会誌. 14. 141-146 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakahata T,Xu M,Ueda T,Matsuoka S,Ito M,Tsuji K: "Cell Therapy"Springer. 3 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 中畑龍俊: "サイトカインの造血幹細胞移植への応用"羊土社. 6 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 中畑龍俊: "サイトカインと疾患1"医学のあゆみ. 5 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Futaki M., Yamashita T., Yabe M., kato S., Nakahata T.: "The IVS4+4A-T mutation of the Fanconi Anemia gene FANCC is not associated with severe phenotype in Japanese patients."Blood. 95. 1493-1498 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sugiyama H., Nonaka T.Komoriya K., Tsuji K., Nakahata T.: "Peroxisome proliferator-activated receptors negatively regurate mast cell activation"FEBS Letters. 467. 259-262 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ueda Y, Tsuii K, Yoshino H, Ebihara Y, Yagasaki H, Yasukawa K, Nakahata T: "Expansion of human NOD/SCID-repopulating cells by a combination of stem cell factor, Flk2/Flt3 ligand, thrombopoietin and a complex of interleukin-6 and soluble interleukin-6 receptor"J Clin Invest. 105. 1013-1021 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hayashi Y., Honma Y., Niitsu N., Tsuji K., Nakahata T.: "SN-1, a novel leukemic cell line with t(11 ; 16)(q23 ; p13) : Myeloid characteristics and resistance to retinoids and vitamin D3."Cancer Res.. 60. 1139-1145 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Misawa K., Nosaka T., Morita S., Nakahata T., Asano S., Kitamura T.: "A method to identify cDNA based on localization of green fluorescent protein fusion products."Proc.Natl.Acad.Sci.USA. 97. 3062-3066 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ebihara Y, Xu M, Nakahata T, Tsuju K: "Exclusive expression of G-CSF receptor on myeloid progenitors in bone marrow CD34+ cells"Br J Haematol. 109. 153-161 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ueda T, Manabe A, Kikuchi A, Yoshino H, Ebihara Y, Ishii T, Tsuji K, Nakahata T: "Massive pericardial and pleural effusion with anasarca following allogeneic bone marrow transplantation"Int J Hematol. 71. 394-397 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ueda T, Yoshino H, Kobayashi K, Asano S, Nakahata T, Tsuji K: "Hematopoietic repopulating ability of cord blood CD34+ cells in NOD/Shi-scid mice"Stem Cells.. 18. 204-213 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Matsui A., Sato T., Maekawa T., Asano S., Nakahata T., Tsuji K.: "Glycoprotein 130 and c-kit signals synergistically induce thrombopoietin production by hematopoietic cells"Int.J.Hematol.. 72. 455-462 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sato T, Maekawa T, Watanabe S, Tsuji K, Nakahata T: "Erythroid progenitors differentiate and mature in response to endogenous erythropoietin"J Clin Invest, .. 106. 263-270 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sasaki S, Ito E, Toki T, Maekawa T, Nakahata T, Igarashi K, Yokoyama M: "Cloning and expression of human B cell-specific transcription factor BACH2 mapped to chromosome 6q15"Oncogene.. 19. 3739-3749 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tanaka Y, Naruse I, Xu M, Nakahata T, Maekawa T, Ishii S: "Extensive brain hemorrhage and embyonic lethality in a mouse null mutant of CREB-binding protein."Mechanisms of Development. 95. 133-145 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishii T, Manabe A, Ebihara Y, Nakahata T, Asano S, Tuji K: "Improvement in bronchiolitis obliterans organizing pneumonia in a child after allogeneic bone marrow transplantation by a combination of oral prednisolone and low dose erythromycin"Bone Marrow Transplantation.. 26. 907-910 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Muroi J, Yorifuji T, Uematsu A, Kitamura A, Nakahata T: "Molecular and clinical analysis of Japanese patients with 3-hydroxy-3-methylglutaryl CoA lyase (HL) deficiency"Hum Genet.. 107. 320-326 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Muroi J, Yorifuji T, Uematsu A, Nakahata T: "Cerebral infarction and pancreatitis : possible complications of patients with 3-hydroxy-3-methylglutaryl-CoA lyase deficiency"J.Inherit.Metab.Dis.. 23. 636-637 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Higuchi Y, Hattori H, Tsuji M, Asato R, Nakahata T: "Partial seizures in leukoencephalopathy with swelling and a discrepantly mild clinical course"Brain & Development.. 22. 387-389 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakahata T.: "Ex vivo expansion of human hematopoietic stem cells"Int J Hematol.. 73(1). 6-13 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshino H, Ueda T, Kawahata M, Asano S, Nakahata T, Tsuji K: "Natural killer cell depletion by ant asialo GM1 antiserum treatment enhances human hematopoietic stem cell engraftment in NOD/Shi-scud mice"Bone Marrow Transplant.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sugiyama H, Nonaka T, Tsuji K, Nakahata T: "Peroxisome proliferator-activated receptors are expressed in mouse bone marrow-derived mast cells"FEBS Lett.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ma F, Yanx F, Kaneko A, Asano S, Nakahata T, Tsuji K: "Cytokine requirement for the development of T-lineage potential from lymphohematopoietic progenitor cells"Br J Haematol. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Matsuoka S, Ebihara Y, Xu M, Ikeda Y, Nakahata T, Tsuji K: "CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Xu M, Matsuoka S, Yang F-C, Asano S, Nakahata T, Tsuji K: "Evidence for the presence of primitive megakaryopoiesis in the early yolk sac"Blood. in press.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamashita T, Nakahayta T.: "Current knowledge on the pathophysiology of fanconi anemia : from genes to phenotypes"Int J Hematol. in press.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nishikawa M., Tahara T., Nakahata T., Shimosaka A.: "Role of the microenvironment of the embryonic aorta-gonad-mesonephros region in hematopoiesis"Stem Cells in press. in press.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sawano A., Iwai S., Sakurai Y., 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. in press.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Matsuoka S, Tsuii K, Ikeda Y, Asano S., Nakahata T: "Generation of definitive hematopoietic stem cells from murine early yolk sac and paraaortic splanchnopleura by AGM region-derived stromal cells"Blood. in press.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ma F., Wada, M., Yoshino, H., Nakahata, T., Asano, S., Tsuji, K.: "Development of human lymphohematopoietic stem/progenitor cells defined by CD34 and CD81 expressions"Blood. in press.

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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