Research of rheumatoid arthritis using Dbl knock-out mice

使用 Dbl 敲除小鼠进行类风湿性关节炎研究

• 批准号: 15591055
• 负责人: HASHIRAMOTO Akira
• 金额: $ 2.3万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591055/
• 关键词: Dbl; Rheumatoid arthritis; 遺伝子除去マウス

Rheumatoid arthritis (RA) is a chronic polyarthritis finally leading to joint destruction. While the ratio of the risk for siblings of patients with a disease was much greater in familial RA, suggesting that genetic factors may be important as a cause of familial clustering, we have previously reported in Japanese familial RA with 3 principal chromosomal regions of linkage, D1S253/214, D8S556 and DXS1232. Komai et al. have subsequently identified DBL proto-oncogene (wild type DBL; GenBank/EMBL/DDBJ Accession No.AB085902) located on DXS1232/984 as a candidate for RA disease gene : a mutant DBL cDNA lacking exons 23 and 24 (deleted type DBL ; GenBank/EMBL/DDBJ Accession No. AB085901) was associated with familial RA.The DBL proto-oncogene is a prototype guanine nucleotide exchange factor (GEF) that modulates activity of small G proteins, including Rho family GTPases, RhoA and Cdc42 and possibly Rac1. Dbl converts the client proteins from the GDP-bound (inactive) form to the GTP-bound (act … More ive) form, allowing its effector domain to interact with downstream signaling molecules. Previous studies have shown that the Rho family GTPases control actin cytoskeleton organization and modulate movement, proliferation, and apoptosis of the cell. In humans, Rho family GEFs play important roles in the maturation and organization of skeletal muscles or nerves, also in the maturation and cytokine production of lymphoid cells. The mutation in some of the GEF family genes has been implicated in the human disease such as faciogenital dysplasia ^<15)>. It have been reported that deleted form of Dbl showed a weaker GEF activity toward Cdc42, and that infiltration and NADPH oxidase activity of neutrophils were significantly decreased in rheumatoid patients with this deleted mutant DBL.In the present study, we established mouse embryonic fibroblast(MEF) cell lines from Dbl knock-out mice and evaluated the activity of Rho family proteins and the intracellular localization and mobility of Dbl protein in mutant or wild type MEFs. The result showed that Dbl proto-oncogene affect the activation of Rho family proteins in MEFs through the decrease of intracellular mobility. We are now going to back-cross Dbl knock-out mice with DBIJ mice to examine the effects of Dbl gene-deletion on collagen-induced arthritis in mice. Less

风湿性关节炎（RA）是一种慢性多发性关节炎，最终导致关节破坏。虽然在家族性RA中，患有疾病的患者的兄弟姐妹的风险比要大得多，这表明遗传因素可能是家族聚集的重要原因，但我们以前曾报道过日本家族性RA有3个主要的染色体连锁区域，D1 S253/214，D8 S556和DXS 1232。Komai等人随后鉴定了DBL原癌基因（野生型DBL; GenBank/EMBL/DDBJ Accession No.AB085902）定位于DXS 1232/984上作为RA疾病基因的候选基因：缺失外显子23和24的突变型DBL cDNA（删除DBL型; DBL原癌基因是一种原型鸟嘌呤核苷酸交换因子（GEF），调节小G蛋白的活性，包括Rho家族GTP酶、RhoA和Cdc 42，可能还有Rac 1。Dbl将客户蛋白从GDP结合（无活性）形式转化为GTP结合（活性）形式。 ...更多信息 iv）形式，允许其效应域与下游信号分子相互作用。以前的研究表明，Rho家族GTP酶控制肌动蛋白细胞骨架的组织，并调节细胞的运动，增殖和凋亡。在人类中，Rho家族GEF在骨骼肌或神经的成熟和组织中起重要作用，也在淋巴细胞的成熟和细胞因子产生中起重要作用。一些GEF家族基因中的突变与人类疾病如面生殖器发育不良有关。已有报道，Dbl缺失型对Cdc 42显示出较弱的GEF活性，并且在具有这种缺失突变体DBL的类风湿患者中，中性粒细胞的浸润和NADPH氧化酶活性显著降低。我们建立了小鼠胚胎成纤维细胞（MEF）细胞系，在小鼠中，评估了Rho家族蛋白的活性以及Dbl蛋白在突变型或野生型MEFs中的细胞内定位和移动性。结果表明，Db 1原癌基因通过降低细胞内迁移率影响MEFs中Rho家族蛋白的活化。我们现在将Dbl基因敲除小鼠与DBIJ小鼠回交，以检查Dbl基因缺失对小鼠胶原诱导的关节炎的影响。少

项目成果

Estrogen specifically stimulates expression and production of osteoprotegerin from rheumatoid synovial fibroblasts.

• DOI: 10.3892/ijmm.15.5.827
• 发表时间: 2005-05
• 期刊: International journal of molecular medicine
• 影响因子: 5.4
• 作者: M. Mitani;Y. Miura;R. Saura;A. Kitagawa;T. Fukuyama;A. Hashiramoto;S. Shiozawa;M. Kurosaka;S. Yoshiya

KAWASAKI H: "Human weel kinase is directly transactivated by and increased in association with c-Fos/AP-1 : rheumatoid synovial cells overexpressing these genes go into aberrant mitosis"Oncogene. 22・44. 6839-6844 (2003)

川崎 H：“人类 Weel 激酶直接被 c-Fos/AP-1 反式激活并增加：过度表达这些基因的类风湿滑膜细胞进入异常有丝分裂”Oncogene 6839-6844。

関節リウマチと分子シャペロン-HSP90を介した滑膜細胞のシグナル伝達系-

类风湿性关节炎和分子伴侣-HSP90介导的滑膜细胞信号转导系统-

• 发表时间: 2004
• 期刊: 臨床リウマチ 16.3
• 作者: Miki;Murata;Osawa K;Murata M;柱本照
• 通讯作者: 柱本照

Heat shock protein 90 (HSP90) is required for increased DNA binding activity of activator protein-1 (AP-1), a heterodimer of Fos/JunD, in rheumatoid synovial cells under inflammatory stimuli.

热休克蛋白 90 (HSP90) 是类风湿滑膜细胞在炎症刺激下增加激活蛋白 1 (AP-1)（Fos/JunD 异二聚体）的 DNA 结合活性所必需的。

• 发表时间: 2005
• 期刊: Int.J.Mol.Med. 15
• 作者: Miki;Murata
• 通讯作者: Murata

Yamashita T: "Enhanced insulin sensitivity in mice lacking ganglioside GM3"Proc Natl Acad Sci U S A. 100・6. 3445-3449 (2003)

Yamashita T：“缺乏神经节苷脂 GM3 的小鼠的胰岛素敏感性增强”Proc Natl Acad Sci U S A. 100・6 (2003)。