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.虽然致白血病酪氨酸激酶激活常见的下游分子，但这些LTK引起的白血病的表型是不同的。在本研究中，我们利用嗜酸性粒细胞增多综合征/慢性嗜酸性粒细胞白血病的致病基因F1P1L1-PDGFRα(F-PRα)分析其机制。当引入 c-Kit^<high>Sca-1^+ 谱系细胞 (KSL) 时，F- 。 PRa而非TEL-PDGFRβ (T-PRβ)增强了Gr-1^<+>IL-5Rα^<+>嗜酸性粒细胞祖细胞(EoPs)的发育。此外，F-PRα促进了常见骨髓祖细胞(CMPs)的嗜酸性粒细胞发育。此外，当在巨核细胞/红细胞祖细胞 (MEP) 和共同淋巴祖细胞 (CLP) 中表达时，关于这一机制，F-PRα 异常地从 MEP 和 CLP 中发育出 EoP。RT-PCR 分析表明，F-PRα 增强了 C/EBPα 和 GATA-2 的表达，同时减少了 PU 下游 Ras 的表达。 2 活性，而它们在荧光素酶测定中抑制 PU.1 活性。2。在这项研究中，我们生成了 AM 转基因（Tg）小鼠，尽管 AM Tg 小鼠没有自发产生任何肿瘤，但由于 B 细胞的生长而观察到了明显的脾肿大。使用免疫组织化学染色，AM 是弥漫性大 B 细胞淋巴瘤特殊亚型的不良预后因素。 3．使用人间充质干细胞（hMSC）作为基质的脐带血，我们可以从 2000 个 CD34_+ 细胞中培养出 1-5 x 10^<5> CD10_+ 细胞。 4周后出现。此外，我们发现激活素A选择性抑制B淋巴细胞的产生。