Analysis of novel chemokine "WHCHE" in development and hematopoies

新型趋化因子“WHCHE”在发育和造血中的分析

• 批准号: 12670997
• 负责人: OHNEDA Osamu
• 金额: $ 2.11万
• 依托单位: University of Tsukuba (2001-2002)Kumamoto University (2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670997/
• 关键词: Chemokine; Endothelial cell; Flk-1; WHCHE(CXCL15); Transgenic mouse; 血管新生; 造血細胞; WECHE; アデノウイルス; 胎仔肝臓; 赤芽球造血; 遺伝子破壊マウス

A novel chemokine 'WECHE' (CXCLI 5) was isolated from a dorsal aorta-derived endothelial stromal cell line. It was reported previously that WECHE inhibits the proliferation of yolk sac-derived endothelial cells and the differentiation of fetal liver-derived hematopoletic cells. In order to investigate a role of WECHE in vivo, WECHE was overexpressed in vivo by using Adenovirus system and transgenic mouse.(1) Adenovirus systemAdenovirus was constructed by the ligation of WECHE cDNA under the controlled of chicken beta-actin promoter. The Adeno-WECHE was infected to E13.5 embryos and fetal liver was analyzed at E17. WECHE infected fetal liver contains two-fold higher number of CFU-E compared to the control (Adeno-LacZ).(2) Transgenit mouseCAAG 'promoter was used to generate WECHE transgenic mouse. Over fifty numbers of mice were analyzed, and it was found that one mouse has the transgene by PCR analysis. Then, several kinds of organs were extracted for mRNA, and WECHE expression was analyzed by RT-PCR. None of WECHE mRNA was observed in the transgenic mouse. We are going to generate Flk-1-WECHE transgenic mouse in order to examine the effect of VVECHE for endothelial cell development in vivo.

一种新的趋化因子‘WECHE’(CXCLI5)是从背主动脉来源的内皮基质细胞系中分离得到的。此前有报道称，WECHE抑制卵黄囊来源的内皮细胞的增殖和胎肝来源的造血细胞的分化。为了研究WECHE在体内的作用，利用腺病毒系统和转基因小鼠在体内进行了WECHE的高效表达。(1)腺病毒系统：在鸡β-肌动蛋白启动子的控制下，连接WECHE基因，构建成重组腺病毒。将重组腺病毒感染E13.5胚胎，在E17胎龄时对胎肝进行分析。(2)利用转基因小鼠CAAG启动子构建了WECHE转基因小鼠，通过对50多只小鼠的分析，发现有1只小鼠表达了WECHE基因。然后提取几种器官的mRNA，用RT-PCR分析WECHE基因的表达。在转基因小鼠中没有检测到WECHE基因的表达。我们将建立Flk-1-WECHE转基因小鼠，以检测VVECHE对体内血管内皮细胞发育的影响。

项目成果

Nomiyama, H.: "Organization of the chemokine genes in the human and mouse major cluster of CC and CXC chemokines : diversificaiton between the two species."Genes and Immunity. 2. 110-113 (2001)

Nomiyama, H.：“人类和小鼠 CC 和 CXC 趋化因子主要簇中趋化因子基因的组织：两个物种之间的多样性。”基因与免疫。

Arai F, Ohneda O, Miyamoto T, Zhang XQ, Suda T.: "Mesenchymal stem cells in perichondrium express activated leukocyte cell adhesion molecule and participate in bone marrow formation."Journal of Experimental Medicine. 195. 1549-1563 (2002)

Arai F、Ohneda O、Miyamoto T、Zhang XQ、Suda T.：“软骨膜中的间充质干细胞表达活化的白细胞细胞粘附分子并参与骨髓形成。”实验医学杂志。

Suzuki N, Ohneda O, Takahashi S, Higuchi M, Mukai H, Nakahata T, Imagawa S, Yamamoto M.: "Erythroid-specific expression of erythropoietin receptor rescued its null mutant mice from lethality."Blood. 100. 2279-2288 (2002)

Suzuki N、Ohneda O、Takahashi S、Higuchi M、Mukai H、Nakahata T、Imakawa S、Yamamoto M.：“促红细胞生成素受体的红细胞特异性表达使其无效突变小鼠免于死亡。”血液。

Miyamoto T: "Bifurcation of osteoclasts and dendritic cells from common progenitors"Blood. 98. 2544-2554 (2001)

宫本 T：“来自共同祖细胞的破骨细胞和树突状细胞的分叉”血液。

Suzuki, N.: "Erythroid-specific expression of erythropoietin receptor rescued its null mutant mice from lethality."Blood. 100. 2279-2288 (2002)

Suzuki, N.：“促红细胞生成素受体的红细胞特异性表达使其无效突变小鼠免于死亡。”血液。