2000 年度実績報告書

造血幹細胞自己複製因子の遺伝子クローニングとその臨床応用

研究課題

研究課題/領域番号	11357008
研究機関	京都大学
研究代表者	中畑龍俊京都大学, 医学研究科, 教授 (20110744)
研究分担者	林英蔚京都大学, 医学研究科, 助手 (40283600) 依藤亨京都大学, 医学研究科, 助手 (60220779) 片村憲司京都大学, 医学研究科, 講師 (40185806)
キーワード	造血幹細胞 / 自己複製因子 / AGM領域 / ストローマ細胞株 / 遺伝子クローニング / mRNA / マウス / 未知遺伝子
研究概要	造血幹細胞が発生、増幅することが明らかとなったマウスの胎生10.5日のマウス胎児aorta-gonad-mesonephros(AGM)領域からストローマ細胞株の樹立を試みた。すでに複数のストローマ細胞株の樹立に世界で初めて成功し、この内の1つ(AGMA9)は、サイトカイン無添加でマウス及びヒトいずれの造血幹細胞の長期にわたる増殖支持能を持つという画期的なものであった。また、同じAGM領域から造血幹細胞増殖支持能を持たないストローマ細胞株(AGMA7)の樹立にも成功した。AGMA9、AGMA7およびOP9などの既知の数種類のストローマ細胞株よりmRNAを抽出し、RT-PCR法などを用いて各種サイトカインmRNAの発現を検討した。AGMA9から取り出したmRNAからはSCF,SDF-1,OSM,IL-6,HGF,MIP-1γ,Gas6,MCP-3などが検出されたが、M-CSF,EPO,TPO,Flk2/Flt3 ligand,MIP-1αなど未分化造血前駆細胞に作用することが知られているサイトカインのmRNAは検出されなかった。マウスSCF,OSM,IL-6を組み合わせてヒトCD34+CD38-細胞を培養したが、造血前駆細胞の増幅は認められなかった。造血幹細胞増殖支持能を持つAGMA9および持たないAGMA7や弱い活性を認めるOP9よりcDNAライブラリーを作成し、signal sequence trap,gene discovery arrayなどを用いて、A9に発現していてA7に発現していないまたは発現が少ない遺伝子を中心にクローニングを行っている。そのような遺伝子として既に200個以上同定され、未知の遺伝子も数十個同定されている。現在それらの機能解析が進行中である。

研究成果

(37件)

すべてその他

すべて文献書誌 (37件)

[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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 Clin Invest. 105. 1013-1021 (2000)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Nakahata T.: "Ex vivo expansion of human hematopoietic stem cells"Int J Hematol.. 73(1). 6-13 (2001)
[文献書誌] 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).
[文献書誌] 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).
[文献書誌] 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).
[文献書誌] 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).
[文献書誌] 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).
[文献書誌] Yamashita T,Nakahata T.: "Current knowledge on the pathophysiology of fanconi anemia : from genes to phenotypes"Int J Hematol. (in press).
[文献書誌] Nishikawa M.,Tahara T.,Nakahata T.,Shimosaka A.: "Role of the microenvironment of the embryonic aorta-gonad-mesonephros region in hemtopoiesis"Stem Cells. (in press).
[文献書誌] 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).
[文献書誌] 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).
[文献書誌] 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).
[文献書誌] 中畑龍俊: "造血幹細胞のex vivo増幅と臨床応用"Biotherapy. 14. 270-275 (2000)
[文献書誌] 中畑龍俊: "ヒトES細胞と再生医療"BIO Clinica. 15(6). 322-326 (2000)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Nakakhata T,Xu M,Ueda T,Matsuoka S,Ito M,Tsuji K: "Cell Therapy"Springer. 3 (2000)
[文献書誌] 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)
[文献書誌] 中畑龍俊: "サイトカインと疾患 1"医学のあゆみ. 5 (2000)

2000 年度 実績報告書

造血幹細胞自己複製因子の遺伝子クローニングとその臨床応用

研究代表者

中畑 龍俊 京都大学, 医学研究科, 教授 (20110744)

研究成果

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] Nakahata T.: "Ex vivo expansion of human hematopoietic stem cells"Int J Hematol.. 73(1). 6-13 (2001)

[文献書誌] 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).

[文献書誌] 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).

[文献書誌] 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).

[文献書誌] 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).

[文献書誌] 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).

[文献書誌] Yamashita T,Nakahata T.: "Current knowledge on the pathophysiology of fanconi anemia : from genes to phenotypes"Int J Hematol. (in press).

[文献書誌] Nishikawa M.,Tahara T.,Nakahata T.,Shimosaka A.: "Role of the microenvironment of the embryonic aorta-gonad-mesonephros region in hemtopoiesis"Stem Cells. (in press).

[文献書誌] 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).

[文献書誌] 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).

[文献書誌] 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).

[文献書誌] 中畑龍俊: "造血幹細胞のex vivo増幅と臨床応用"Biotherapy. 14. 270-275 (2000)

[文献書誌] 中畑龍俊: "ヒトES細胞と再生医療"BIO Clinica. 15(6). 322-326 (2000)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] Nakakhata T,Xu M,Ueda T,Matsuoka S,Ito M,Tsuji K: "Cell Therapy"Springer. 3 (2000)

[文献書誌] 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)

[文献書誌] 中畑龍俊: "サイトカインと疾患 1"医学のあゆみ. 5 (2000)

2000 年度実績報告書

中畑龍俊京都大学, 医学研究科, 教授 (20110744)