Synthetic Analysis on the mechanisms of survival and differentiation of hematopoietic cells

造血细胞存活和分化机制的综合分析

基本信息

批准号：
18209034
负责人：
KANAKURA Yuzuru
金额：
$ 28.87万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-18209034/
关键词：
tyrosine kinase apootosis hematopoietic stem cell

项目摘要

1. Although leukemogenic tyrosine kinases activate common downstream molecules, the phenotypes of leukemia caused by these LTKs are distinct. In this study, we analyzed its mechanism using F1P1L1-PDGFRα(F-PRα), a causative gene of hypereosinophilic syndrome/chronic eosinophilic leukemia. When introduced into c-Kit^<high>Sca-1^+ Lineage cells (KSLs), F- PRa but not TEL-PDGFRβ (T-PRβ) enhanced the development of Gr-1^<+>IL-5Rα^<+> eosinophil progenitors(EoPs). Also, F- PRα promoted eosinophil development from common myeloid progenitors (CMPs). Furthermore, when expressed in megakaryocyte/erythrocyte progenitors (MEPs) and common lymphoid progenitors (CLPs), F-PRα aberrantly developed EoPs from MEPs and CLPs. Regarding this mechanism, RT-PCR analysis revealed that F-PRα augmented the expression of C/EBPα and GATA-2, while it reduced PU. 1 expression. Furthermore, F-PRα and its downstream Ras enhanced GATA- 2 activity, while they inhibited PU.1 activity in luciferase assays.2. We previously cloned a novel anti-apoptotic gene, Anamorsin(AM). In this study, we generated transgenic(Tg) mice for AM. Although AM Tg mice did not develop any tumors spontaneously, marked splenomegaly due to the outgrowth of B cells was observed. In addition, we found that the expression of AM was a poor prognostic factor for the special subtype of diffuse large B-cell lymphoma using immunohistochemical staining.3. We developed a long-term culture system to produce B lymphocytes from human CD34_+ cells purified from umbilical cord blood using human mesenchymal stem cells (hMSC) as stroma. Using this cocultures, we could develop 1-5 x 10^<5> CD10_+ cells from 2000 CD34_+ cells. In this system, surface IgM_+ immature B cells began to appear after 4 weeks. In addition, we found that Activin A selectively suppressed B lymphocyte production.

1。尽管白血病酪氨酸激酶激活了共同的下游分子，但这些LTK引起的白血病的表型却截然不同。在这项研究中，我们使用F1P1L1-PDGFRα（F-PRα）分析了其机制，这是一种严重的嗜酸性综合征/慢性嗜酸性白血病的严重基因。当引入c-kit^<高> SCA-1^+谱系细胞（KSLS）时，F-PRA，而不是Tel-PDGFRβ（T-PRβ）增强了GR-1^<+> IL-5Rα^<+>嗜酸性嗜酸性粒细胞元素（EOPS）的发展。同样，F-PRα促进了普通髓样祖细胞（CMP）的嗜酸性粒细胞发育。此外，当在巨核细胞/红细胞祖细胞（MEP）和普通淋巴祖细胞（CLPS）中表达时，F-PRα异常从MEP和CLPS产生了EOPS。关于这种机制，RT-PCR分析表明，F-PRα增强了C/EBPα和GATA-2的表达，而它降低了PU。 1表达。此外，F-PRα及其下游RAS增强了GATA-2活性，而它们抑制了PU.1荧光素酶测定中的活性。2。我们以前以一种新型的抗凋亡基因Anamorsin（AM）克隆。在这项研究中，我们为AM生成了转基因（TG）小鼠。尽管AM TG小鼠并未自发发育任何肿瘤，但由于B细胞的生长而显着脾肿大。此外，我们发现AM的表达是使用免疫组织化学染色的弥漫性大B细胞淋巴瘤特殊亚型的预后因素不良。3。我们开发了一种长期培养系统，以使用人间充质干细胞（HMSC）作为基质从脐带血纯化的人CD34_+细胞产生B淋巴细胞。使用这种共培养，我们可以从2000 CD34_+细胞中开发1-5 x 10^<5> CD10_+细胞。在该系统中，表面IgM_+未成熟的B细胞在4周后开始出现。此外，我们发现活化素A活化蛋白有选择地抑制的B淋巴细胞产生。