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Regulation of self-renewal of hematopoietic stem cells by bone marrow stromal cells

骨髓基质细胞对造血干细胞自我更新的调控

基本信息

批准号：
16390274
负责人：
NAKAJIMA Hideaki
金额：
$ 9.09万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390274/
关键词：
Hematopoietic stem cell Bone marrow stromal cell Self-renewal m Kirre ISF ShIF 骨髄間質細胞ストローマ immune suppressor factor

项目摘要

In order to analyze m Kirre function in vivo, we set out to disrupt m Kirre in mouse genome. We screened BAC library to obtain genomic fragment containing m Kirre gene and isolated several genomic fragments. Restriction mapping of m Kirre genomic fragment combined with the computer data base search completed the genomic map of m Kirre locus. Targeting vector was designed to replace the first exon and large portion of the 5-promoter region with Neomycin cassette to disrupt m Kirre gene. Completed targeting vector was electroporated into E14 embryonic stem (ES) cells, and approximately 400 Neomycin-resistant clones were selected and processed to identify homologously recombinant clones by Southern hybridization. We obtained one correctly targeted clone, which was confirmed by hybridizing with 5'- and 3-probes, together with Neo-probe. We are currently making chimeric mice by injecting this clone into blastcysts from C57BL/6 mice.As for ISF, we performed cDNA microarray analysis of bone marrow stromal cell lines overexpressing ISF and ShIF. Comparing expression profiles of MS10 cells expressing ISF, ShIF or mock plasmids revealed that TIMP-3 and SFRP-1 were down-regulated by ISF overexpression. By re-expressing TIMP-3 or SFRP-1 genes in MS10/ ISF cells, the enhanced hematopoietic stem cell (HSC) supporting activity of MS10/ ISF cells reduced a level similar to that of parental MS10 cells. These results indicate that TIMP-3 and SFRP-1 play critical roles in enhanced HSC activity conferred by ISF.

为了分析m Kirre在体内的功能，我们开始破坏小鼠基因组中的m Kirre。我们筛选BAC文库获得含有m Kirre基因的基因组片段，并分离出多个基因组片段。m Kirre基因组片段的限制性内切图谱结合计算机数据库检索完成m Kirre位点的基因组图谱。设计靶向载体，用新霉素盒取代第一个外显子和大部分5启动子区域，破坏m Kirre基因。将完整的靶向载体电穿孔到E14胚胎干细胞中，选择约400个新霉素耐药克隆，并通过Southern杂交鉴定同源重组克隆。我们获得了一个正确的靶向克隆，通过与5'和3-探针以及neo -探针杂交证实了这一点。我们目前正在通过将这种克隆注射到C57BL/6小鼠的囊胚中来制造嵌合小鼠。对于ISF，我们对过表达ISF和shiff的骨髓基质细胞系进行了cDNA芯片分析。比较表达ISF、shift或模拟质粒的MS10细胞的表达谱发现，ISF过表达下调了TIMP-3和SFRP-1。通过在MS10/ ISF细胞中重新表达TIMP-3或SFRP-1基因，增强的MS10/ ISF细胞造血干细胞（HSC）支持活性降低了与亲代MS10细胞相似的水平。这些结果表明TIMP-3和SFRP-1在ISF增强HSC活性中起关键作用。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Integrin αIIbβ3 induces the adhesion and activation of mast cells through interaction with fibrinogen

整合素αIIbβ3通过与纤维蛋白原相互作用诱导肥大细胞粘附和活化

DOI：
发表时间：
2006
期刊：
J.Immunol. 176
影响因子：
0
作者：
Baba T;Mimura J;Nakamura N;Harada N;Yamamoto M;Morohashi K;Fujii-Kuriyama Y.;沖俊彦;中島秀明;中島秀明;福地由美;川島敏行;沖俊彦
通讯作者：
沖俊彦

Dimerization of MLL fusion proteins and FLT3 activation synergize to induce multiple-lineage leukemogenesis