Molecular mechanisms regulating the growth and differentiation of hematopoietic stem cells

调节造血干细胞生长和分化的分子机制

• 批准号: 12470200
• 负责人: KANAKURA Yuzuru
• 金额: $ 8.83万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470200/
• 关键词: ras; proliferation; differentiation; signal transduction; STAT; transcription factor; megakaryocyte; AIM-1

(1) Stem cell factor (SCF) has crucial roles in proliferation, survival and differentiation of hematopoietic stem cells through binding to c-Kit receptor (KIT). We made a series of 22 KIT mutants, in which Tyr (Y) residue was substituted to Phe (F) in the cytoplasmic domain, and introduced into BAF3 cells. On stimulation with SCF, BAF3 expressing KIT^<WT>(WT) showed cell migration and Ca^<2+> mobilization. Among 22YF mutants, Y567F and Y719F showed significantly reduced cell migration and Ca^<2+> mobilization. Analysis on signaling cascades suggested that Y567-mediated Src family kinase (SFK) activation led to Ca^<2+> influx and migration, and that P38MAPkinase (P38MAPK) and Erk1/2 were also regulated by Y567/SFK and involved in cell migration. Also, Y719-mediated PI3K pathway was suggested to be involved in the migration. These results indicate that two major KIT signaling pathways lead to cell migration, one is Y567-SFK-p38MAPK-Erk and another is Y719-PI3 kinase.(2) Thrombopoietin (T … More PO) and its receptor c-mpl play crucial roles in growth and megakaryocytic differentiation of hematopoietic stem cells. We found that Ras activation was involved in thrombopoietin (TPO)-induced megakaryocytic differentiation. Furthermore, we demonstrated that GATA-1 activities was required for the Ras-mediated megakaryocytic differentiation, and that GATA-1 activities were regulated negatively by the direct interaction with other lineage-specific transcription factors, PU.1 and c-Myb.(3) AIM-1 belongs to an Aurora/Ipl1 serine threonine kinase family, and is supposed to play key roles in mitosis. In human hematopoietic cells, expression of AIM-1 was restrictedly observed at G2/M phase of cell cycle. In contrast, AIM-1 was continuously repressed during megakaryocytic polyploidization. Supplement of AIM-1 activities by the induced expression of wild-type AIM-1 canceled TPA-induced polyploidization of K562 cells, and the suppression of AIM-1 activities by dominant-negative AIM-1 led to polyploidization. These results suggested that down-regulation of AIM-1 at M phase may be involved in abortive mitosis and polyploid formation of megakaryocytes. Less

（1）干细胞因子（SCF）通过与C-KIT接收器（KIT）的结合在造血干细胞的增殖，存活和分化中具有至关重要的作用。我们制作了一系列22个试剂盒突变体，其中Tyr（Y）居住在细胞质结构域中的PHE（F），并引入BAF3细胞中。在用SCF刺激时，表达Kit^<wt>（WT）的BAF3显示了细胞迁移和Ca^<2+>动员。在22年突变体中，Y567F和Y719F显示出细胞迁移和动员的细胞迁移显着降低。对信号级联的分析表明，Y567介导的SRC家族激酶（SFK）激活导致CA^<2+>影响和迁移，并且p38mapkinase（p38mapk）和ERK1/2也受到Y567/SFK的调节，并参与细胞迁移。另外，建议Y719介导的PI3K途径参与迁移。这些结果表明，两种主要的套件信号通路导致细胞迁移，一个是Y567-SFK-P38MAPK-ERK，另一个是Y719-PI3激酶。（2）血栓植物（T…更多PO）及其接收器C-MPL在生长和巨核细胞中的血液中起着至关重要的作用。我们发现RAS激活参与血小板蛋白（TPO）引起的巨核细胞分化。此外，我们证明了RAS介导的巨核细胞分化需要GATA-1活性，并且与与其他特定谱系特异性转录因子PU.1和C-MYB的直接相互作用（3）AIM-1属于AURORA/IPL1柔软的Terreon kinase Kiners，以及对MIT的键入，对GATA-1的活性受到负面调节。在人造血细胞中，在细胞周期的G2/m期有限观察到AIM-1的表达。相反，在巨核细胞多倍化过程中，AIM-1连续反射。通过诱导的野生型AIM-1的表达来补充AIM-1活性，取消了TPA诱导的K562细胞的多倍体化，以及通过显性阴性的AIM-1抑制AIM-1活性导致多倍体化。这些结果表明，在M期的AIM-1下调可能与巨核细胞的流产有丝分裂和多倍体形成有关。较少的

项目成果

Kawasaki, A., Matsumura, I., Miyagawa, J., Ezoe, S., Tanaka, H., Terada, Y., Tatsuka, M., Machii, T., Miyazaki, H., Furukawa, Y., and Kanakura, Y.: "Down-regulation of and AIM-1 kinase couples with megakaryocytic polyploidization of human hematopoietic ce

川崎，A.，松村，I.，宫川，​​J.，Ezoe，S.，田中，H.，寺田，Y.，龙香，M.，町井，T.，宫崎，H.，古川，Y.，

Nishimura, J-I., Philips, K.L., Ware, R.E., Hall, S., Wilson, L., Gentry, T.L., Howard, T.A.., Murakami, Y., Shibano, M., Machii, T., Gilboa, E., Kanakura, Y., Takeda, J., Kinoshita, T., Rosse, W.F., and Smith, C.A.: "Efficient retrovirus-mediated PIG-A g

Nishimura, J-I.、Philips, K.L.、Ware, R.E.、Hall, S.、Wilson, L.、Gentry, T.L.、Howard, T.A.、Murakami, Y.、Shibano, M.、Machii, T.、Gilboa, E

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Ueda, S., et al.：“c-kit 酪氨酸残基 567 和 719 在干细胞因子诱导的趋化性中的关键作用”血液。

Matsumura, I., Hashimoto, K, Ikeda, H., Odajima, J., Tanaka, H., Kato, T, Miyazaki, H., and Kanakura, Y.: "Increased D-type cyclin expression together with decreased cdc2 activity confers megakaryocytic differentiation of a human thrombopoietin-dependent

Matsumura, I.、Hashimoto, K、Ikeda, H.、Odajima, J.、Tanaka, H.、Kato, T、Miyazaki, H. 和 Kanakura, Y.：“D 型细胞周期蛋白表达增加，同时 cdc2 减少

Sonoyama, J., et al.: "Functional cooperation among Ras, STAT5, and P13-K is required for full oncogenic activitics of BCR/ABL in K562 cells"J. Biol. Chem.. (in press).

Sonoyama, J. 等人：“K562 细胞中 BCR/ABL 的完全致癌活性需要 Ras、STAT5 和 P13-K 之间的功能合作”。