Attempt to induce hematoopoietic stem cells from hemangioblasts

尝试从成血管细胞诱导造血干细胞

基本信息

批准号：
11470206
负责人：
HARA Takahiko
金额：
$ 8.64万
依托单位：
Tokyo Metropolitan Organization For Medical Research, The Tokyo Metropolitan Institute of Medical Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470206/
关键词：
AGM region hemangioblast PCLP1 AML1 c-myb hematopoietic stem cells endothelial cells stem cell plasticity Runix-1

项目摘要

Hematopoietic stem cells are known to be derived from hemangioblasts, common precursor cells for blood and endothelial cells. We have identified podocalyxin-like protein 1 (PCLP1) as a novel hemangioblast marker. PCLP1+CD45-cells in the AGM region are differentiated to hematopoietic cells and endothelial cells in vitro. When these cells were transplanted into busulfan-treated neonatal mice, they gave rise to hematopoietic stem cells (HSCs) as well as endothelial cells, smooth muscle cells, and stromal cells in lung, small intestine, and uterus. By retrovirus marking of stem cells in the AGM region, we proved that blood cells and vascular cells in small intestine are derived from single stem cells. Similar in vivo differentiation capacity was detected in the bone marrow CD45-positive side population, which are highly enriched for HSCs. Therefore, it is likely that hemangioblasts in the AGM region generated HSCs that are in turn transdifferentiated to non-hematopoietic cells in vivo.We have shown that two transcription factors, AMI-1 and c-myb, are essential for the generation of HSCs in the AGM region. To know functions of these proteins, we newly established immortalized cell lines from the AGM regions of knockout mice ofAMI-1 or c-myb genes. Then we reintroduced missing genes by taking advantage of inducible promoter. Gene expression studies revealed that mRNA for insulin-like growth factor binding protein-3 is suppressed by AML1.Further search of down stream effectors of AML-1 and c-myb would clarify the molecular mechanism of HSC development from hemangioblasts in the AGM region.

已知造血干细胞源自成血管细胞，这是血液和内皮细胞的常见前体细胞。我们已经鉴定出足萼蛋白样蛋白 1 (PCLP1) 作为一种新型成血管细胞标记物。 AGM 区域中的 PCLP1+CD45-细胞在体外分化为造血细胞和内皮细胞。当这些细胞被移植到白消安治疗的新生小鼠体内时，它们会在肺、小肠和子宫中产生造血干细胞（HSC）以及内皮细胞、平滑肌细胞和基质细胞。通过逆转录病毒标记AGM区域的干细胞，我们证明了小肠中的血细胞和血管细胞来源于单个干细胞。在骨髓 CD45 阳性侧群体中检测到类似的体内分化能力，该群体高度富集 HSC。因此，AGM区域的成血管细胞很可能产生HSC，然后在体内转分化为非造血细胞。我们已经证明，AMI-1和c-myb这两种转录因子对于AGM区域中HSC的产生至关重要。年度股东大会区域。为了了解这些蛋白质的功能，我们从 AMI-1 或 c-myb 基因敲除小鼠的 AGM 区域新建立了永生化细胞系。然后我们利用诱导型启动子重新引入缺失的基因。基因表达研究表明，胰岛素样生长因子结合蛋白-3的mRNA被AML1抑制。进一步寻找AML-1和c-myb的下游效应子将阐明AGM区域成血成血管细胞发育HSC的分子机制。