Receptor-mediated signalings-regulating proliferation, differentiation and neoplastic transformation of hematopoietic cells

受体介导的信号传导——调节造血细胞的增殖、分化和肿瘤转化

• 批准号: 09470231
• 负责人: KANAKURA Yuzuru
• 金额: $ 9.09万
• 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: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470231/
• 关键词: c-kit; c-mpl; tyrosine kinase; thrombopoietin; signal transduction; proliferation; differentiation; ras; ras; 造血因子; 受容体; 増殖; 分化; 腫瘍

The c-kit receptor tyrosine kinase (KIT) and its ligand, stem cell factor (SCF) transduce crucial signals in hematopoietic cells. We revealed that two point mutations, Val559→Gly (G559) mutation in the juxtamembrane domain and Asp814→Val (V814) mutation in the kinase domain, lead to constitutive and oncogenic activation of KIT. In order to determine as to which portion of mutant KITs is indispensable for receptor dimerization or self-association in the absence of SCF, we have constructed a series of deletion and chimeric mutants of KIT, including the extracellular domain truncated type KIT and chimeric type KIT in which extracellular and transmembrane domains are replaced by src myristylation signal peptide. By using murine interleukin (IL)-3-dependent Ba/F3 cells that were subjected to transfection of various c-kit genes, we showed that the constituvely activating mutations at both the juxtamembrane and kinase domains of KIT may not necessarily require their extracellular and transmem … More brane domains for their formation of receptor self-association, and that the receptor self-association of mutant KIT may be important for activation of downstream effectors that are required for factor-independent growth and tumorigenicity.In addition to the SCF/KIT system, thrombopoietin (TPO)/c-mpl system also plays a fundamental role in hematopoiesis. In an effort to clarify the mechanisms of TPO-induced proliferation and differentiation, c-mpl was introduced into human IL-3-dependent F-36P cells. F-36P-mpl cells were found to proliferate and differentiate at a high rate into mature megakaryocytes in response to TPO. By using dominant-negative (dn) forms of STATs and ras, it was suggested that TPO-induced proliferation may be mediated through activation of STAT5 and ras, and that prolonged ras activation may be involved in TPO-induced megakaryocytic differentiation. Furthermore, we found that STAT5, in addition to ras signaling, appeared to mediate transcriptional regulation of cyclin D1 during cytokine-dependent growth in hematopoietic cells. Less

c-kit受体酪氨酸激酶（KIT）及其配体干细胞因子（SCF）在造血细胞中传递关键信号。我们发现两个点突变，即近膜结构域的Val559→Gly （G559）突变和激酶结构域的Asp814→Val （V814）突变，导致KIT的组成性和致癌激活。为了确定在没有SCF的情况下，哪一部分突变体KIT对于受体二聚化或自结合是必不可少的，我们构建了一系列KIT的缺失和嵌合突变体，包括细胞外结构域截断型KIT和嵌合型KIT，其中细胞外和跨膜结构域被src肉肉酰基化信号肽取代。通过使用转染各种c-kit基因的小鼠白细胞介素(IL)-3依赖性Ba/F3细胞，我们发现KIT近膜域和激酶域的组构激活突变可能不一定需要它们的细胞外和跨膜域。突变KIT的受体自关联可能对下游效应物的激活很重要，而下游效应物是不依赖因子生长和致瘤性所必需的。除了SCF/KIT系统外，血小板生成素(TPO)/c-mpl系统在造血中也起着重要作用。为了阐明tpo诱导的增殖和分化机制，将c-mpl引入人il -3依赖性F-36P细胞。发现F-36P-mpl细胞在TPO作用下以高速率增殖和分化为成熟的巨核细胞。通过对STATs和ras的显性阴性（dn）表达，我们发现tpo诱导的巨核细胞增殖可能是通过STAT5和ras的激活介导的，而ras的长时间激活可能参与了tpo诱导的巨核细胞分化。此外，我们发现STAT5，除了ras信号外，似乎在造血细胞细胞因子依赖性生长过程中介导cyclin D1的转录调节。少

项目成果

Tsujimura, T. , et al.: "Consitutively activating mutation in the catalytic domain of c-kit elicits hematopoietic transformation by receptor self-association not at the ligand-dependent dimerization site"Blood. 93. 1319-1329 (1999)

Tsujimura, T. 等人：“c-kit 催化结构域中的组成性激活突变可通过受体自缔合而不是在配体依赖性二聚化位点引发造血转化”Blood。

Nishida,T., et al.: "Familial gastrointestinal stromal tumors with germ line mutation of the KIT gene." Nature Genet.19. 323-324 (1998)

Nishida,T., et al.：“具有 KIT 基因种系突变的家族性胃肠道间质瘤。”

Hirota,S., et al.: "Gain-of-function mutations of c-kit in human gastrointestinal stromal tumors." Science. 279. 577-580 (1998)

Hirota,S. 等人：“人胃肠道间质瘤中 c-kit 的功能获得突变。”

Matsumura,I., et al.: "Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiation of a human factor-dependent hematopoietic cell line." Mol.Cell.Biol.18. 4282-4290 (1998)

Matsumura,I. 等人：“ras 延长激活参与血小板生成素诱导的人类因子依赖性造血细胞系的巨核细胞分化。”

Matsumura, I., et al.: "Thrombopoietin-induced differentiation of a human megakaryoblastic leukemia cell line, CMK, involves transcriptional activation of p21ィイD1WAF1/Cip1ィエD1 by STAT5"Mol. Cell. Biol. 17. 2933-2943 (1997)

Matsumura, I., et al.：“血小板生成素诱导的人巨核细胞白血病细胞系 CMK 的分化涉及 STAT5 对 p21D1WAF1/Cip1D1 的转录激活”，《细胞生物学》。