サイトカインによるシグナル伝達機構の解析

细胞因子诱导的信号转导机制分析

• 批准号: 07409001
• 负责人: MIYAJIMA Atsushi
• 金额: $ 23.36万
• 依托单位: Institute of Molecular and Cellular Biosciencesa, The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07409001/
• 关键词: cytokine; cytokine receptors; signal transduction; hematopoiesis; differentiation; apoptosis; RAS; STAT; サイトカイン; サイトカイン受容体; 血管内皮; 発生; 誘導遺伝子; 細胞分化; 造血因子; JAK; Ras; 幹細胞; 造血細胞; 増殖; 細胞死; キナーゼ; 細胞増殖

As studies on signaling by using the mutant IL-3/GM-CSF/IL-5 receptors revealed two major signaling pathways, RAS and STAT5, we have focused our efforts to elucidate the role of these pathways in cytokine functions. Hematopoietic cells require cytokines for their growth and also for their survival. We found that activation of RAS is important for suppression of cell death. RAS activates both RAF/MAP kinase and P13 kinase and both pathways were found to be involved in suppression of cell death. We also showed that caspase-3 is involved in cell death induced by cytokine depletion in hematopoietic cells. To find the role of STAT5 in cytokine functions, we isolated STAT5 target genes. Among them, ClS encodes a novel SH2 protein that negatively regulates cytokine signaling. Expression of a dominant negative form of STAT5 blocked proliferation induced by IL-3 and erythroid differentiation induced by EPO, indicating that STAT5 plays a distinct role depending on the cell tynes. Cross talk between STAT5 and RAS was also demonstrated. In addition, we studied the function of Oncostatin M(OSM), another STAT5 target gene, and found that OSM plays an important role in development of hematopoietic cells and germ cells. The OSM receptor cDNA was cloned and the molecular structure of the functional receptor was elucidated.

由于使用突变型IL-3/GM-CSF/IL-5受体进行的信号转导研究揭示了两种主要的信号转导途径，RAS和STAT 5，因此我们集中精力阐明这些途径在细胞因子功能中的作用。造血细胞需要细胞因子用于其生长和存活。我们发现RAS的激活对于抑制细胞死亡是重要的。RAS激活RAF/MAP激酶和P13激酶，发现这两种途径都参与抑制细胞死亡。我们还表明，caspase-3参与了造血细胞中细胞因子耗尽诱导的细胞死亡。为了发现STAT 5在细胞因子功能中的作用，我们分离了STAT 5靶基因。其中，ClS编码一种新的SH 2蛋白，负调节细胞因子信号传导。STAT 5的显性负性形式的表达阻断了由IL-3诱导的增殖和由EPO诱导的红系细胞分化，表明STAT 5根据细胞类型发挥不同的作用。STAT 5和RAS之间的串扰也被证明。此外，我们还研究了STAT 5的另一个靶基因Oncostatin M（OSM）的功能，发现OSM在造血细胞和生殖细胞的发育中起重要作用。克隆了OSM受体cDNA，并阐明了功能性受体的分子结构。

项目成果

Chida, D.et al.: "Role of cytokine signaling molecules in erythrold defferentiation of mouse fetal liver hematopoietic cells : functional analvsis of signaling molecules by retrovirus-mediated gene expression." Blood. (in press). (1999)

Chida, D.等人：“细胞因子信号分子在小鼠胎肝造血细胞红细胞分化中的作用：通过逆转录病毒介导的基因表达对信号分子进行功能分析。”

Mui,A.L-F.et.al: "Interleukin-3,granulocyte-macrophage colony stimulating factor,and interleukin-5 transduce signals through two STAT5 homologues" EMBO J.14. 1166-1175 (1995)

Mui，A.L-F.et.al：“白细胞介素 3、粒细胞巨噬细胞集落刺激因子和白细胞介素 5 通过两个 STAT5 同源物转导信号”EMBO J.14。

Wakao, H., Harada, N., Kitamura, T., Mui, A.L., and Miyajima, A.: "Interleukin-2 and erythropoietin activate STAT5 via distinct pathways." EMBO J.14. 2527-2535 (1995)

Wakao, H.、Harada, N.、Kitamura, T.、Mui, A.L. 和 Miyajima, A.：“Interleukin-2 和促红细胞生成素通过不同的途径激活 STAT5。”

Furusawa, T. et al.: "Integrin-associated protein (IAP, also termed CD47) is involved in stroma-supported erythropoiesis." J. Biochem.123. 101-106 (1998)

Furusawa, T. 等人：“整合素相关蛋白（IAP，也称为 CD47）参与基质支持的红细胞生成。”

Kinoshita, T., M.Shirouzu, A.Kamiya, K.Hashimoto, S.Yokoyama and A.Miyajima: "Raf/MAPK-and rapamycin-sensitive pathways mediate the anti-apoptotic function of p21Ras in IL-3 dependent hematopoietic cells." Oncogene. 15. 619-627 (1997)

Kinoshita, T., M.Shirouzu, A.Kamiya, K.Hashimoto, S.Yokoyama 和 A.Miyajima：“Raf/MAPK 和雷帕霉素敏感途径介导 IL-3 依赖性造血细胞中 p21Ras 的抗凋亡功能