MOLECULAR BASIS OF STEM CELL SYSTEMS AND ITS APPLICATION

干细胞系统的分子基础及其应用

• 批准号: 14207006
• 负责人: NAKANO Toru
• 金额: $ 28.54万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14207006/
• 关键词: cell differentiation; stem cell; hematopoiesis; transcription factor; embryonic stem cell; macrophage; erythrocyte; megakaryocyte; 生殖細胞; 転写調節

In order to reveal the molecular differentiation mechanisms in stem cell systems, we established a novel experimental strategy combining an ES cell in vitro differentiation method (OP9 system) and Tet-off conditional gene expression method. Using the system, we analyzed the functions of transcriptional factors and co-factors involved in hematopoietic differentiation such as GATA-1, GATA-2, Runx-1 and FOG-1.During the course of hematopoietic differentiation in the presence of M-CSF (macrophage-colony stimulating factor), macrophages are preferentially produced. When a hematopoietic zinc finger protein, GATA-2, is over-expressed, macrophage differentiation is blocked and differentiation switch to erythroid or megakaryocytic lineages occurs. And the determination to. erythroid cells or megakaryocytes were determined by the timing of GATA-2 expression. In addition, TSA (trichostatin A), an HDAC (histone de-acetylase) inhibitor skewed the differentiation from megakaryocyte to erythroid lineage. Further analyses revealed that expression level of PU.1, another hematopoietic transcription factor, played crucial roles for the lineage determination to erythroid or megakaryocytic lineages.Now the functions of the other transcription factors are under investigation.

为了揭示干细胞系统的分子分化机制，我们建立了一种结合ES细胞体外分化方法（OP9系统）和et-off条件基因表达方法的新实验策略。利用该系统，我们分析了GATA-1、GATA-2、Runx-1和FOG-1等参与造血分化的转录因子和辅助因子的功能。在造血分化过程中，巨噬细胞集落刺激因子（M-CSF）的存在优先产生巨噬细胞。当造血锌指蛋白GATA-2过表达时，巨噬细胞分化受阻，分化向红系或巨核细胞谱系转变。和决心。红细胞或巨核细胞通过GATA-2表达的时间来确定。此外，TSA (trichostatin A)，一种HDAC（组蛋白去乙酰化酶）抑制剂扭曲了巨核细胞向红系的分化。进一步的分析表明，另一种造血转录因子PU.1的表达水平对红系或巨核细胞谱系的确定起着至关重要的作用。现在其他转录因子的功能正在研究中。

项目成果

Suzuki A, Kaisho T, (6 authors), Nakano T.: "Critical roles of Pten in B cell homeostasis and immunoglobulin class switch recombination"J Exp Med. 197. 657-667 (2003)

Suzuki A、Kaisho T（6 位作者）、Nakano T.：“Pten 在 B 细胞稳态和免疫球蛋白类别转换重组中的关键作用”J Exp Med。

Kuramochi-Miyagawa K, Kimura T, (10 authors), Nakano T: "mili, a mammalian member of piwi family gene, is essential for spermatogenesis"Development. 131. 839-849 (2004)

Kuramochi-Miyakawa K，Kimura T，（10位作者），Nakano T：“mili，piwi家族基因的哺乳动物成员，对于精子发生至关重要”的发展。

Suzuki, A, Kaisho, T., Ohishi, M., Tsukio-Yamaguchi, M., Tsubata T., Koni, P.A., Sasaki, T., Mak, T.W., Nakano, T。: "Critical roles of Pten in B cell homeostasis and immunoglobulin class switch recombination."J.Exp.Med.. 197. 657-667 (2003)

铃木，A，Kaisho，T.，Ohishi，M.，Tsukio-Yamaguchi，M.，Tsubata T.，Koni，P.A.，Sasaki，T.，Mak，T.W.，Nakano，T.

Masuhara M, Nagao K, Nishikawa M, Kimura T, Nakano T: "Enhanced degradation of MDM2 by a nuclear envelope component, mouse germ cell less"Biochem Biophys Res Commun. 308. 927-932 (2003)

Masuhara M、Nagao K、Nishikawa M、Kimura T、Nakano T：“核膜成分增强 MDM2 的降解，小鼠生殖细胞较少”Biochem Biophys Res Commun。

Nakano T: "Hematopoietic stem cells : generation and manipulation"Trends Immunol. 24. 589-594 (2003)

Nakano T：“造血干细胞：生成和操作”趋势免疫学。