1999 Fiscal Year Annual Research Report

ヒト造血幹細胞の解析と白血病発症機構の研究

Research Project

Project/Area Number	11139215
Research Institution	Kyoto University
Principal Investigator	中畑龍俊京都大学, 医学研究科, 教授 (20110744)
Co-Investigator(Kenkyū-buntansha)	依藤亨京都大学, 医学研究科, 助手 (60220779) 辻浩一郎東京大学, 医科学研究所, 助教授 (50179991)
Keywords	G-CSF / G-CSF受容体(G-CSFR) / トランスジェニックマウス / 造血幹細胞 / 血球分化 / Kostmann症候群 / 好中球減少症 / 白血病
Research Abstract	G-CSFRからの細胞増殖シグナル伝達には、細胞内領域のbox1,box2をふくむN端側が、分化シグナル伝達にはN末端とC末端の両者が必要であることが報告されている.先天性好中球減少症であるKostmann症候群の一部の患者ではG-CSFR遺伝子の変異によりG-CSFRの細胞内領域のC末端の欠失が見られ、これが患者で見られる好中球系前駆細胞の成熟障害の直接的原因であろうと考えられている。近年、G-CSFを長期使用されているKostmann症候群患者にMDSや急性白血病が高頻度に合併することが明らかとなり、G-CSFRの変異と急性白血病発症との関係が注目されるとともに臨床的に大きな問題となっている。本研究ではwild typeおよびC末端を欠失した変異G-CSFRを発現するTgマウスを作製し、変異G-CSFRを有する個体へのG-CSFの長期投与が急性白血病発症の原因となるか否か検討することを主目的としている。また、G-CSFRを介する刺激による造血幹細胞の増殖・分化過程を検討すると共に、2種類のG-CSFRからの細胞内シグナル伝達の違いを明らかにし、白血病発症との関係を解析することを目的としている。Wild typcおよびKostmann症候群患者で見られるG-CSF受容体(G-CSFR)遺伝子のmutationを持ち、C末端を欠失した2種類の変異G-CSFRトランスジェニック(Tg)マウスを作成した。Wild typeのG-CSFR Tgマウスにおいて、末梢血中の血球数に変化はなく、G-CSFは骨髄細胞から好中球、マクロファージ、好酸球、巨核球、肥満細胞、芽球など様々な細胞の増殖・分化・成熟を刺激した.このようなG-CSFの作用はclone-sortingした細胞の単細胞培養でも確認された。以上の結果より、血球分化におけるサイトカイン受容体シグナルの特異性は極めて低いことが示唆された。Kostmann症候群患者で見られるG-CSFR遺伝子のmutationを持ち、C末端を欠失した2種類の変異G-CSFR-Tgマウスの作成に成功した.いずれにおいても著明な好中球減少が見られており、世界で始めてKostmann症候群のモデルマウスが作成できたと考えられる。現在、好中球減少機序の詳細な検討及びこれらのマウスへのhG-CSFのin vivo投与により、白血病、MDSが惹起されるか否か検討している。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] Sui X., Tsuji K., Ebihara Y., TKishimoto T., Nakahata T.: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34+ cells through gp130 signaling"Blood. 93. 2525-2532 (1999)
[Publications] Hasegawa S., Pawankar R., Suzuki K., Nakahata T., Furukawa S., Okumura K., Ra C: "Functional expression of the high affinity receptor for IgE (Fc epsilon RI) in human Platelets and its intracellular expression in human megakaryocytes"Blood. 93. 2543-2551 (1999)
[Publications] Honda H., Suzuki T., Inaba T., Tsuji K., Nakahata T., Yazaki Y., Look T., Hirai H.: "Expression of E2a-HLF chineric protein induced T-cell apotosis, B-cell maturation arrest, and development of acute lymphoblastic leukemia"Blood. 93. 2780-2790 (1999)
[Publications] Kampurai D., Saito H., Kaneko A., Fukagawa K., Tsuji K., Nakahata T.: "Characterization of mast-cell-committed progenitors present in human cord blood"Blood. 93. 3338-3346 (1999)
[Publications] Kobayashi M., Ueda K., Kojima S., Shimbo T., Nishihira H., Nakahata T.: "Serum granulocyte colony-stimulating factor levels in patients with chronic neutropenia of childhood : Modulation of G-CSF levels by myeloid precursor cell mass"Brit. J. Haematol.. 105. 486-490 (1999)
[Publications] Taniguchi, T., Endo H., Asano S>, Fujita T., Nakahata T., Motokura T.: "Expression of p21 Cip1/Waf1/Sdi1 and p27 kip1 cyclin-dependent kinase inhibitors during human hematopoiesis"Blood. 83. 4167-4178 (1999)
[Publications] Kinoshita T., Sekiguchi T., Xu M-J., Ito Y., Tsuji K., Nakahata T., Miyajima A.: "Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis"Proc. Natl. Acad. Sci. USA. 96. 7265-7270 (1999)
[Publications] Yang FC., Tsuji K., Oda A., Xu MJ., Ebihara Y., Watanabe S., Ikeda Y., Nakahata T.: "Effects of human granulocyte colony-stimulating factor (hG-CSF) on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice"Blood. 94. 950-9958 (1999)
[Publications] Matsumoto T., Seki Y., Kubo M., Nakahata T., Yamada S., Yoshimura A.: "Suppression of STAT5 functions in liver, mammary grands, and T cells in cytokine-inducible SH2 protein-1 (CIS1) transgenic mice"Mol. Cell. Biol.. 19. 6396-6407 (1999)
[Publications] Ui M., Takeda M., Arai T., Yamada K., Nakahata T., Tokino T., Nakamura Y., Iba H.: "Retrovirus vectors designed for efficient transduction of cytotoxic or cytostatic genes"Gene Therapy. 6. 1670-1678 (1999)
[Publications] Ishii T., Nakahata T., Maekawa T.: "Expression of SDF-1/PBSF receptor CXCR4 on CD34+ human bone marrow cellsis a phenotypic alteration for committed lymphoid progenitors"J. Immunol.. (in press).
[Publications] Kajiwara M., Nonoyama S., Eguchi M., Imai K., Ochs HD., Nakahata T., Yata J.: "In vitro megakaryopocyte differentiation from CD34 positive cells is defective in patients with Wiscott-Aldrich syndrome and X-linked thrombocytopenia"Brit. J. Haematol.. (in press).
[Publications] Takagi M., Nakamura T., Sawada T., Nakahata T., Yokota T., Heike T.: "Chimeric M-CSF-leukemia inhibitory factor (LIF) receptor can transduce expansion signals in primitive hematopoietic progenitors lacking IL-6Ra, IL-11Ra and gp130"Mol. Cell. Biol.. (in press).
[Publications] Ishiguro A., Nakahata T., Kato T., Suzuki Y., Shimbo T.: "Age-related changes in serum thrombopoietin in children : high levels in infants by one month of age"Pediatr, Res.. (in press).
[Publications] Futaki M., Yamashita T., Yabe M., Kato S., Asano S., Nakahata T.: "The IVS4+4A-T mutation of the Fanconi Anemia Gene FANCC is not associated with severe phenotype in Japanese patients"Blood. (in press).
[Publications] Matsuoka S., Tsuji K., Xu M., Yang FC., Kaneko K., Kikuchi A., Nakahata T.: "CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).
[Publications] Ueda T., Yoshino H., Ebihara Y., Ito M., Tsuji K., Nakahata T.: "Ex vivo expansion of human NOD/SCID-repopulating cells by stem cell factor, Flk2/Flt3 ligand, thrombopoietin interleukin-6 and soluble interleukin-6 receptor"J. Clin. Invest.. (in press).
[Publications] Hayashi Y., Honma Y., Nitsu N., Taki T., 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.. (in press).
[Publications] Sugiyama H., Nonaka T., Kishimoto T., Komoriya K, Tsuji K., Nakahata T.: "Peroxisome proliferator-activated receptors negatively regurate mast cell activation"FEBS Letters. (in press).
[Publications] Sugiyama H., Nonaka T., Kishimoto T., Komoriya K., Tsuji K., Nakahata T.: "Peroxisome proliferator-activated receptors are expressed in human cultured mast cells : a possible role for these receptors in negative regulation of mast cell activation"Blood. (in press).
[Publications] 中畑龍俊: "In vitroでの末梢血幹細胞の増幅. 症例に学ぶG-CSFの臨床"医薬ジャーナル. 14 (1999)
[Publications] 中畑龍俊、二木真琴: "遺伝子治療開発研究ハンドブック"日本遺伝子治療学会編、エヌ・テイ・エス. 6 (1999)
[Publications] 辻浩一郎、中畑龍俊: "内科学教科書"文光堂(東京)(印刷中).

1999 Fiscal Year Annual Research Report

ヒト造血幹細胞の解析と白血病発症機構の研究

Principal Investigator

中畑 龍俊 京都大学, 医学研究科, 教授 (20110744)

Research Products

[Publications] Sui X., Tsuji K., Ebihara Y., TKishimoto T., Nakahata T.: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34+ cells through gp130 signaling"Blood. 93. 2525-2532 (1999)

[Publications] Hasegawa S., Pawankar R., Suzuki K., Nakahata T., Furukawa S., Okumura K., Ra C: "Functional expression of the high affinity receptor for IgE (Fc epsilon RI) in human Platelets and its intracellular expression in human megakaryocytes"Blood. 93. 2543-2551 (1999)

[Publications] Honda H., Suzuki T., Inaba T., Tsuji K., Nakahata T., Yazaki Y., Look T., Hirai H.: "Expression of E2a-HLF chineric protein induced T-cell apotosis, B-cell maturation arrest, and development of acute lymphoblastic leukemia"Blood. 93. 2780-2790 (1999)

[Publications] Kampurai D., Saito H., Kaneko A., Fukagawa K., Tsuji K., Nakahata T.: "Characterization of mast-cell-committed progenitors present in human cord blood"Blood. 93. 3338-3346 (1999)

[Publications] Kobayashi M., Ueda K., Kojima S., Shimbo T., Nishihira H., Nakahata T.: "Serum granulocyte colony-stimulating factor levels in patients with chronic neutropenia of childhood : Modulation of G-CSF levels by myeloid precursor cell mass"Brit. J. Haematol.. 105. 486-490 (1999)

[Publications] Taniguchi, T., Endo H., Asano S>, Fujita T., Nakahata T., Motokura T.: "Expression of p21 Cip1/Waf1/Sdi1 and p27 kip1 cyclin-dependent kinase inhibitors during human hematopoiesis"Blood. 83. 4167-4178 (1999)

[Publications] Kinoshita T., Sekiguchi T., Xu M-J., Ito Y., Tsuji K., Nakahata T., Miyajima A.: "Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis"Proc. Natl. Acad. Sci. USA. 96. 7265-7270 (1999)

[Publications] Yang FC., Tsuji K., Oda A., Xu MJ., Ebihara Y., Watanabe S., Ikeda Y., Nakahata T.: "Effects of human granulocyte colony-stimulating factor (hG-CSF) on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice"Blood. 94. 950-9958 (1999)

[Publications] Matsumoto T., Seki Y., Kubo M., Nakahata T., Yamada S., Yoshimura A.: "Suppression of STAT5 functions in liver, mammary grands, and T cells in cytokine-inducible SH2 protein-1 (CIS1) transgenic mice"Mol. Cell. Biol.. 19. 6396-6407 (1999)

[Publications] Ui M., Takeda M., Arai T., Yamada K., Nakahata T., Tokino T., Nakamura Y., Iba H.: "Retrovirus vectors designed for efficient transduction of cytotoxic or cytostatic genes"Gene Therapy. 6. 1670-1678 (1999)

[Publications] Ishii T., Nakahata T., Maekawa T.: "Expression of SDF-1/PBSF receptor CXCR4 on CD34+ human bone marrow cellsis a phenotypic alteration for committed lymphoid progenitors"J. Immunol.. (in press).

[Publications] Kajiwara M., Nonoyama S., Eguchi M., Imai K., Ochs HD., Nakahata T., Yata J.: "In vitro megakaryopocyte differentiation from CD34 positive cells is defective in patients with Wiscott-Aldrich syndrome and X-linked thrombocytopenia"Brit. J. Haematol.. (in press).

[Publications] Takagi M., Nakamura T., Sawada T., Nakahata T., Yokota T., Heike T.: "Chimeric M-CSF-leukemia inhibitory factor (LIF) receptor can transduce expansion signals in primitive hematopoietic progenitors lacking IL-6Ra, IL-11Ra and gp130"Mol. Cell. Biol.. (in press).

[Publications] Ishiguro A., Nakahata T., Kato T., Suzuki Y., Shimbo T.: "Age-related changes in serum thrombopoietin in children : high levels in infants by one month of age"Pediatr, Res.. (in press).

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

[Publications] Matsuoka S., Tsuji K., Xu M., Yang FC., Kaneko K., Kikuchi A., Nakahata T.: "CD34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).

[Publications] Ueda T., Yoshino H., Ebihara Y., Ito M., Tsuji K., Nakahata T.: "Ex vivo expansion of human NOD/SCID-repopulating cells by stem cell factor, Flk2/Flt3 ligand, thrombopoietin interleukin-6 and soluble interleukin-6 receptor"J. Clin. Invest.. (in press).

[Publications] Hayashi Y., Honma Y., Nitsu N., Taki T., 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.. (in press).

[Publications] Sugiyama H., Nonaka T., Kishimoto T., Komoriya K, Tsuji K., Nakahata T.: "Peroxisome proliferator-activated receptors negatively regurate mast cell activation"FEBS Letters. (in press).

[Publications] Sugiyama H., Nonaka T., Kishimoto T., Komoriya K., Tsuji K., Nakahata T.: "Peroxisome proliferator-activated receptors are expressed in human cultured mast cells : a possible role for these receptors in negative regulation of mast cell activation"Blood. (in press).

[Publications] 中畑龍俊: "In vitroでの末梢血幹細胞の増幅. 症例に学ぶG-CSFの臨床"医薬ジャーナル. 14 (1999)

[Publications] 中畑龍俊、二木真琴: "遺伝子治療開発研究ハンドブック"日本遺伝子治療学会編、エヌ・テイ・エス. 6 (1999)

[Publications] 辻浩一郎、中畑龍俊: "内科学教科書"文光堂(東京)(印刷中).

中畑龍俊京都大学, 医学研究科, 教授 (20110744)