造血細胞のシグナル伝達

• Project/Area Number: 10181101
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 宮島 篤 東京大学, 分子細胞生物学研究所, 教授 (50135232)
• Co-Investigator(Kenkyū-buntansha): 吉村 昭彦 九州大学, 生体防御医学研究所, 教授 (90182815) ; 田賀 哲也 熊本大学, 発生医学研究センター, 教授 (40192629) ; 福永 理己郎 大阪大学, 医学部, 助教授 (40189965) ; 金倉 譲 大阪大学, 医学部, 教授 (20177489) ; 横田 崇 金沢大学, 医学系研究科, 教授 (50134622) ; 小松 則夫 自治医科大学, 講師 (50186798)
• Project Period (FY): 1998 – 2001
• Project Status: Completed (Fiscal Year 2001)
• Budget Amount: ¥207,200,000 (Direct Cost: ¥207,200,000); Fiscal Year 2001: ¥53,800,000 (Direct Cost: ¥53,800,000); Fiscal Year 2000: ¥52,600,000 (Direct Cost: ¥52,600,000); Fiscal Year 1999: ¥50,400,000 (Direct Cost: ¥50,400,000); Fiscal Year 1998: ¥50,400,000 (Direct Cost: ¥50,400,000)
• Keywords: 造血因子 / サイトカイン / シグナル伝達 / 受容体 / キナーゼ / 転写因子 / 細胞増殖 / 細胞分化

Research Abstract

造血細胞の増殖分化に関わるシグナルリングについての本年度の主要な成果は以下のようである。
AGM領域で発生した造血幹細胞胎生肝臓での増幅をin vitroで再現し、長期の造血再構築能が発生段階が進むにつれて低下することを示した(宮島)。gp130欠損マウスマウスのAGM培養では、血球の出現がほぼ完全に抑制され、AGM内の背側大動脈に発現する分子としてendomucin-1を同定し、この分子が細胞接着阻害作用をもつことを見いだした(田賀)。c-Kitを介する細胞遊走シグナルやCa2+influxには、Y567/srcファミリーキナーゼ/p38MAPK経路とY719/PI3kinase経路の二経路が重要であることを明らかにした(金倉)。c-Kitに結合する新規分子Spredをクローニングし,細胞膜に局在し活性化されたRas, Rafと複合体を形成することで、チロシンキナーゼからのRas/MAKキナーゼ経路を選択的に阻害することを明らかにした(吉村)。Stat1とStat3の造血における機能を解析するために、抑制型Stat3トランスジェニックマウス、さらにStat1のKOマウスとの交配により、抑制型Stat3/Stat1 KOマウスを作製し、Stat3は巨核球系前駆細胞の増幅に、Stat1は赤芽球系前駆細胞の増幅に重要であることを示した(小松)。骨髄球特異的転写因子MZF-2に結合する核内因子を探索し,SWI2/SNF2型クロマチン再構成因子であるmDomino/p400を同定し,骨髄球分化に関わる遺伝子発現には,MZF-2とmDominoの相互作用によるクロマチン再構成が関与することを示唆した(福永)。新規Znフィンガータンパク質SalIのノックアウトマウスでは左右の腎臓が完全に欠失していた。胎生11.5日に尿管芽が間葉に進入して後腎の発生は開始するが、ノックアウトマウスではこの段階で発生が障害されており、この遺伝子は後腎発生のもっとも初期に必須であることが判明した。この遺伝子は腎臓未分化細胞である後腎間葉のみならず、神経幹細胞、ES細胞でも発現しており、共通の機構が幹細胞において働いている可能性が示唆された(横田)。

Research Products

• [Publications] Matsui T. et al.: "K-Ras mediates cytokine-induced formation of adherens junctions during liver development"EMBO J.. 21. 1021-1030 (2002)
• [Publications] Takeuchi M. et al.: "Cultivation of AGM-derived hematopoietic stem cells in the fetal liver microenvironment amplifies long-term repopulating activity and enhances homing to the bone marrow"Blood. 99. 1190-1196 (2002)
• [Publications] Kinoshita T. et al.: "Oncostatin M suppresses generation of progenitors in fetal liver by inhibiting the hepatic microenvironment"Experimental Hematology. 29. 1091-1097 (2001)
• [Publications] Tanaka, M. et al.: "Forkhead family transcription factor FKHRL1 is expressed in human megakaryocytes and regulates cell cycling as a downstream molecule of TPO signaling"J. Biol. Chem.. 276. 15082-15089 (2001)
• [Publications] Kirito, K. et al.: "Identification of the human erythropoietin receptor region required for Stat1 and Stat3 activation"Blood. 99. 102-110 (2002)
• [Publications] Kirito, K. et al.: "A Functional role of Stat3 in in vivo megakaryopoiesis"Blood. (in press). (2002)
• [Publications] Kirito, K. et al.: "TPO regulates Bcl-xL gene expression through Stat5 and phosphatidylinositol-3-kinase activation pathways"J. Biol. Chem.. (in press). (2002)
• [Publications] Fukurnoto, T. et al.: "The Fused protein kinase regulates Hedgehog-stimulated transcriptional activation in Drosophila Schneider 2 cells"J. Biol. Chem.. 276. 38441-38448 (2001)
• [Publications] Kawasaki, A. et al.: "Down-regulation of an AIM-1 kinase couples with megakaryocytic polyploidization of human hematopoietic cells"J. Cell Biol.. 152. 275-287 (2001)
• [Publications] Nojima, J. et al.: "Anti-prothrombin antibodies combined with lupus anticoagulant activity is an essential risk factor for venous thrombolism in patients with systemic lupus erythrmatosus"Br. J. Haematol. 114. 647-654 (2001)
• [Publications] Ueda, S. et al.: "Critical roles of c-kit tyrosine residues 567 and 719 in stem cell factor-induced chemotaxis : contribution of src family kinase and PI3-kinase on calcium mobilization and cell migration"Blood. (in press). (2002)
• [Publications] Sonoyama, J. et al.: "Functional cooperation among Ras, STAT5, and PI3-K is required for full oncogenic activities of BCR/ABL in K562 cells"J. Biol. Chem.. (in press). (2002)
• [Publications] Mizuki, M. et al.: "Role of receptor tyrosine kinases in hematological malignancies"Res. Adv. Blood. (in press). (2002)
• [Publications] Takizawa, T. et al.: "Directly linked soluble IL-6 receptor-IL-6 fusion protein induces astrocyte differentiation from neuroepithelial cells via activation of STAT3"Cytokine. 13. 272-279 (2001)
• [Publications] Nakashima, K. et al.: "BMP2-mediated alteration in the developmental pathway of fetal mouse brain cells from neurogenesis to astrocytogenesis"Proc. Natl. Acad. Sci. USA. 98. 5868-5873 (2001)
• [Publications] Ochiai, W. et al.: "Astrocyte differentiation of fetal neuroepithelial cells involving cardiotrophin-1-induced activation of stat3"Cytokine. 14. 264-271 (2001)
• [Publications] Takizawa, T. et al.: "DNA methylation is a critical cell-intrinsic determinant of astrocyte differentiation in the fetal brain"Dev. Cell. 1. 749-758 (2001)
• [Publications] Nishinakanmra, R. et al.: "Murine homolog of SALL1 is essential for ureteric bud invasion in kidney development"Development. 128. 3105-3115 (2001)
• [Publications] Wakioka, T. et al.: "Spred, a Sprouty-related suppressor of Ras signaling"Nature. 412. 647-651 (2001)
• [Publications] Shouda, T. et al.: "Induction of the cytokine signal regulator SOCS3/CIS3 as a therapeutic strategy for treating inflammatory arthritis"J. Clin. Invest.. 108. 1781-1788 (2001)
• [Publications] Hanada, T. et al.: "A mutant form of JAB/SOCS1 augments the cytokine-induced JAK/STAT pathway by accelerating degradation of wild-type JAI3/CIS family proteins through the SOCS-box"J. Biol. Chem.. 276. 40746-40754 (2001)
• [Publications] Sasaki, A. et al.: "Identification of a dominant negative form of Sproutys that potentiates FGF-but not EGF-induced ERK activation"J. Biol. Chem.. 276. 36804-36808 (2001)
• [Publications] Suzuki, A. et al.: "CIS3/SOCS3/SSI3 Plays a Negative Regulatory Role in STAT3 Activation and Intestinal Inflammation"J. Exp. Med.. 193. 471-482 (2001)
• [Publications] Kamizono, S. et al.: "The SOCS box of SOCS-1 accelerates ubiquitin-dependent proteolysis of TEL-JAK2"J. Biol. Chem.. 276. 12530-12538 (2001)
• [Publications] Minehata, K. et al.: "Macrophage-colony stimulating factor modulates the development of hematopoiesis by stimulating the differentiation of endothelial cells in the AGM region"Blood. (in press). (2002)
• [Publications] Ito Y. et al.: "Retroviral gene transfer of signaling molecules into murine fetal hepatocytes defines distinct roles for the STAT3 and Ras pathways during hepatic development."Hepatology. 32. 1370-1376 (2000)
• [Publications] Kojima N. et al.: "Cell-density-dependent regulation of hepatic development by a gp 130-independent pathway"Biochem.Biophys.Res.Commun. 277. 152-158 (2000)
• [Publications] Miyajima A. et al.: "Roles of Oncostatin M in hematopoiesis and liver development."Cytokine and Growth Factor Reviews. 11. 177-183 (2000)
• [Publications] Sato A. et al.: "Cloning and expression pattern of a Xenopus pronephros-specific gene, XSMP-30"Mechanism of Development. 92. 273-275 (2000)
• [Publications] Nakayama,N. et al.: "A novel chordin-like protein inhibitor for bone morphogenetic proteins"Blood.. (in press). (2001)
• [Publications] Matsumura,I. et al.: "Increased D-type cyclin expression together with decreased cdc2 activity confers megakaryocytic differentiation of a human thrombopoietin-dependent hematopoietic cell line"J.Biol.Chem.. 275. 5553-5559 (2000)
• [Publications] Sakamoto H. et al.: "Mitogen-Activated Protein Kinase Plays an Essential Role in the Erythropoietin-Dependent Proliferation of CTLL-2 Cells"J.Biol.Chem.. 275. 35857-35862 (2000)
• [Publications] Sasaki A. et al.: "CIS3/SOCS3 suppresses erythropoietin signaling by binding the EPO receptor and JAK2"J.Biol.Chem.. 275. 29338-29347 (2000)
• [Publications] Yasukawa H. et al.: "Negative Regulation of Cytokine Signaling Pathways"Annu.Rev.Immunol.. 18. 143-164 (2000)
• [Publications] Ward AC. et al.: "The jak-stat pathway in normal and perturbed hematopoiesis"Blood. 95. 19-29 (2000)
• [Publications] M.Yanagisawa et al.: "Astrocyte differentiation of fetal neuroepithelial cells by interleukin-11 via activation of a common cytokine signal transducer, gpl30, and a transcription factor, STAT3"J.Neurochem. 74. 1498-1504 (2000)
• [Publications] N.Kimura et al.: "BMP2-induced apoptosis is mediated by activation of TAK1-p38 kinase pathway that is negatively regulated by Smad6"J.Biol.Chem.. 275. 17647-17652 (2000)
• [Publications] M.Yanagisawa et al.: "Signaling crosstalk underlying synergistic induction of astrocyte differentiation by BMPs and IL-6 family of cytokines"FEBS Lett.. 24546. 1-5 (2001)
• [Publications] Kashii,Y. et al.: "A member of forkhead family transcription factor FKHRL1 is one of the downstream molecule of phosphatidylinositol 3-kinaze-Akt activation pathway in erythropoietin signal transduction"Blood. 96. 941-949 (2000)
• [Publications] Tanaka,H. et al.: "GATA-1 blocks interleukin-6 induced macrophage differentiation and apoptosis through the sustained expression of cyclin D1 and bcl-2 in a murine myeloid cell line M1"Blood. 95. 1264-1273 (2000)
• [Publications] Daino,H. et al.: "Induction of apoptosis by extracellular ubiquitin in human hematopoietic cells : possible involvement of Stat3 degradation by proteasome pathway in interleukin 6-dependent hematopoietic cells."Blood. 95. 2577-2585 (2000)
• [Publications] Odajima,J. et al.: "Full oncogenic activities of v-Src are mediated by cooperation of multiple signaling pathways ; ras as an essential mediator for cell survival"J.Biol.Chem.. 275. 24096-24105 (2000)
• [Publications] Matsumura,I. et al.: "Biological significance of GATA-1 activities in ras-mediated megakaryocytic differentiation of hematopoietic cell lines"Blood. 96. 2440-2450 (2000)
• [Publications] Mizuki,M. et al.: "Transformation of 32D cells by Flt3 mutations from AML patients is mediated by the Ras And STAT5 pathways"Blood. 96. 3907-3914 (2000)
• [Publications] Kawasaki,A. et al.: "Down-regulation of an AIM-1 kinase couples with megakaryocytic polyploidization of human hematopoietic cells"J.Cell Biol. 152. 275-287 (2001)
• [Publications] Suzuki A. et al.: "CIS3/SOCS3/SSI3 Plays a Negative Regulatory Role in STAT3 Activation and Intestinal Inflammation"J.Exp.Med.. (in press). (2001)
• [Publications] Kamizono S. et al.: "The SOCS box of SOCS-1 accelerates ubiquitin-dependent proteolysis of TEL-JAK2"J.Biol.Chem.. (in press). (2001)
• [Publications] Uchida,M. et al.: "A functional role of mitogen-activated protein kinaze, Erk1 and Erk2, in the differentiation of a human leukemia cell line UT-7/GM : a possible key factor for cell fate determination toward erythroid and megakaryocytic lineages"Int.Haematol.. 73. 78-83 (2001)
• [Publications] Kirito,K. et al.: "Identification of a novel Stat1 and Stat3 activation motif within the erythropoietin receptor"Blood.. (in press). (2001)
• [Publications] Tanaka,M. et al.: "Forkhead family transcription factor FKHRL1 regulates cell cycle of megakaryocytic cells as a downstream molecule of TPO signaling"J.Biol.Chem.. (in press). (2001)
• [Publications] Hefner,Y. et al.: "Serine 727 Phosphorylation and Activation of Cytosolic Phospholipase A2 by MNK1-related Protein Kinases"J.Biol.Chem.. 275. 37542-37551 (2000)
• [Publications] Kinoshita T. et al.: "Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis."Proc. Natl. Acad. Sci. U.S.A.. 96. 7265-7270 (1999)
• [Publications] Hara T. et al.: "Identification of podocalyxin like protein 1 as a novel cell surface marker for hemangioblasts in the murine aorta-gonad-mesonephros region"Immunity. 11. 567-578 (1999)
• [Publications] Nakayama K. et al.: "The novel Oncostatin M-Inducible Gene OIG37 Forms a Gene Family with MyD118 and GADD45 and Negative Regulates Cell Growth"J. Biol. Chem.. 274. 24766-24772 (1999)
• [Publications] Matusda, T. et al.: "STAT3 activation is sufficient to maintain an undifferentiated state of mouse embrvonic stem cells."EMBO J.. 18. 4261-4269 (1999)
• [Publications] Takagi, M. et al.: "Chimeric cytokine receptor can transduce expansion signals in interleukin 6 receptor a (IL-6Ra), IL-11Ra-, and gp 130 low to -negative primitive hematopoietic progenitors"Mol Biol Cell. 10. 3633-3642 (1999)
• [Publications] Nishinakamura, R. et al.: "Activation of Stat3 by cytokine receptor gp130 ventralizes Xenopus embryos independent of BMP-4"Dev Biol.. 216. 481-490 (1999)
• [Publications] Nakashima, K. et al.: "Synergistic signaling in fetal brain by STAT3- Smad1 complex bridged by p300"Science. 284. 479-482 (2000)
• [Publications] Matsumura, I.. et al.: "Increased D-type cyclin expression together with decreased cdc2 activity confers megakaryocytic differentiation of human thrombopoietin-dependent hematopoietic cell line."J. Biol. Chem.. 275. 5553-5559 (2000)
• [Publications] Tanaka, H. et al.: "GATA-1 blocks interleukin-6 induced macrophage differentiation and apoptosis through the sustained expression of cyclin D1 and bcl-2 in a murine myeloid cell line M1"Blood. 95. 1264-1273 (2000)
• [Publications] Tominaga, S. et al.: "Presence and expression of a novel variant form of ST2 gene production in human leukemic cell line UT-7/GM"Biochem. Biophs. Res. Commum.. 264. 14-18 (1999)
• [Publications] Kashii, Y. et al.: "A member of forkhead family transcription factor FKHRL1 is one of the downstream molecule of phosphatidylinositol 30 kinase-Akt activation pathway in erythropoietin signal transduction"Blood in press.
• [Publications] Ward AC. et al.: "The jak-stat pathway in normal and perturbed hematopoiesis"Blood. 95. 19-29 (2000)
• [Publications] Marine JC. et al.: "SOCS3 Is Essential in the Regulation of Fetal Liver Erythropoiesis"Cell. 98. 617-627 (1999)
• [Publications] Marine JC et al.: "SOCS1 Deficiency Causes a Lymphocyte-Dependent Perinatal Lethality"Cell. 98. 609-616 (1999)
• [Publications] Mitsui K. et al.: "Cloning and Characterization of a Novel p21 cip1/Waf1-Interacting Zinc Finger Protein, Ciz1"Biochem. Biophs. Res. Commum.. 264. 457-464 (1999)
• [Publications] Matsumoto A. et al.: "Suppression of STAT5 Functions I Liver, Mammary Glands, and T Cells in Cytokine-Inducible SH2-Containing Protein 1 Transgenic Mice"Mol. Cell Biol.. 19. 6396-6407 (1999)
• [Publications] Sasaki A. et al.: "Cytokine-inducible SH2 protein-3 (CIS3/SOCS3) inhibits Janus tyrosine kinase by binding through the N-terminal kinase main."Genes to Cells. 4. 339-351 (1999)
• [Publications] Suzuki,R.et al.: "ICIS3 and JAB have different regulatory roles in interleukin-6 mediateddifferentiation and STAT3 activation in M1 leukemia cells" Oncogene. 17. 2271-2278 (1998)
• [Publications] Kirito,K.et al.: "A novel function of Statla and Stat3 in the erythropoletin-induced erythroid differentiation of a human leukemla cell line" Blood.92. 412-417 (1998)
• [Publications] Itoh,T.et al.: "Definition of the role of tyrosine residues of the common b subunit regulating multiple signaling pathwavs of granulocvte-macrophage colonv-stimulating factor receptor" Mol.Cell.Biol.18. 742-752 (1998)
• [Publications] Matsmura,I.et al.: "Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiation of a human factor-dependent hematopoletic cell line" Mol.Cell.Biol.18. 4282-4290 (1998)
• [Publications] Tsujimura,T.et al.: "Constitutively activating mutation in the catalytic domain of c-kit receptor tyrosine kinase elicits hematopoletic transformation by receptor self-asociation not at the ligand-dependent dimerization site" Blood. 93. 1319-1329 (1999)
• [Publications] Matsumura,I.et al.: "Transcriptional regulation of cyclin D1 promoter by STAT5:its involvement in cytokine-dependent growth of hematopoletic cells" EMBO J.in press. (1999)
• [Publications] Jiang,W.et al.: "R.PRC1:a human mitotic spindle-associated CDK substrate protein required for cytokinesis" Mol.Cell. 2. 877-885 (1998)
• [Publications] X.-J.Wang.et al.: "gp130,the cytokine common signal-transducer of interleukin-6 cytokine family,is down-regulated in T cells in vivo by interleukin-6" Blood. 91. 3308-3314 (1998)
• [Publications] Mukouyama,Y.et al.: "In vitro expansion of murine hematopoietic progenitors derived from the embryonic aorta gonad mesonephros region." immunity. 8. 105-114 (1998)
• [Publications] Tanaka,M.et al.: "Reconstitution of the functional mouse oncostatin M(OSM)receptot:molecular cloning of the mouse OSM receptor b subunit." Blood. 93. 804-815 (1999)
• [Publications] Chida,D.et al.: "Role of cytokine signaling molecules in erythroid differentiation of mouse fetal liver hematopoietic cells:functional analvsis of signaling molecules by retrovirus-mediated gene expression." Blood in press. (1999)
• [Publications] Yasukawa,H.et al.: "The JAK-Binding Protein JAB Inhibits Janus Tyrosine Kinase Activity Through Binding in the Activation Loop" EMBO J.in press. (1999)
• [Publications] Yokouchi,M.et al.: "APS,an adaptor protein containing PH and SH2 domains,is associated with the PDGF receptor and c-Cb1,and inhibits PDGF-induced mitogenesis" Oncogene in press. (1999)
• [Publications] Sakamoto,H.et al.: "A Janus kinase inhibitor,JAB,is an interferon-g inducible gene andconfers resistance to interferons" Blood. 92. 1668-1676 (1998)