Development of a novel retrovirus vector system for hematopoietic Stem cells

新型造血干细胞逆转录病毒载体系统的开发

• 批准号: 09671106
• 负责人: ITOH Katsuhiko
• 金额: $ 2.05万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671106/
• 关键词: gene therapy; hematopoietic stem cell; retrovirus; レトロウイルス・ベクター; 遺伝子導入

The aims of the project was as follows ;1. Evaluation of the gene expression by a novel retroviral vector which consisted of the SFFVp LTR and the MESV leader sequences.2. Establishment and the evaluation of novel packaging cells based on stromal cells.3. Evaluation of the pseudo-typed retroviral vectors based on SFFVp with VSV G-protein.We have obtained the following results ;1. A series of retroviral vectors were constructed. Using the packaging cells based on NIH/3T3, these vector-producing cells were isolated. Expression levels of these vectors were compared in hematopoietic progenitors. The highest expression of a marker gene among these was obtained by the FMEV-type vector, which contained the SFFVp LTR and the MESV leader sequences.2. Plasmid DNAs of the expression vector carrying gal/pol sequences and that carrying the MoMLV env sequences were introduced into a stromal cell line, MS-5. These novel stromal cell-based packaging cells were found to be unstable to produce high titer of viral vectors.3. SFFVp based retroviral vectors could be pseudo-typed with VSV G-protein and these particles successfully transduced the cells.

该项目的目的如下：1.利用由SFFVp LTR和MESV前导序列组成的新型逆转录病毒载体评价基因的表达。基于基质细胞的新型包装细胞的建立及评价。基于VSV G蛋白的SFFVp伪型逆转录病毒载体的评价：1.构建了一系列逆转录病毒载体。利用基于NIH/3T3的包装细胞，分离出这些产生载体的细胞。比较这些载体在造血祖细胞中的表达水平。其中一个标记基因的最高表达是通过FMEV型载体获得的，该载体含有SFFVp LTR和MESV前导序列。将携带GAL/POL序列的表达载体和携带MoMLV env序列的表达载体分别导入基质细胞系MS-5。这些新型基质细胞包装细胞不稳定，不能产生高滴度的病毒载体。基于SFFVp的逆转录病毒载体可以用VSV G蛋白进行伪分型，这些颗粒成功地转导了细胞。

项目成果

Nishiyama H: "A glycine-rich RNA-binding Protein mediating cold-inducible supression of mammation cell growth" J.Cell Biol. 137. 899-908 (1997)

Nishiyama H：“一种富含甘氨酸的 RNA 结合蛋白介导冷诱导抑制乳腺细胞生长”J.Cell Biol。

Xue JH et al.: "Induction of apg-1,a member of the heat shock protein 110 family,following transient forebrain ischemia in the rat brain" Biochem.Biophys.Res.Commun.247. 796-801 (1998)

薛 JH 等人：“大鼠大脑短暂前脑缺血后，热休克蛋白 110 家族成员 apg-1 的诱导”Biochem.Biophys.Res.Commun.247。

Itoh K: "The role of soluble growth factors in inducing Transient growth and clonal extinction of stroma cell-dependent erythroblastic leukemia cells" Leukemia. 11. 1753-1761 (1997)

Itoh K：“可溶性生长因子在诱导基质细胞依赖性红细胞白血病细胞短暂生长和克隆消亡中的作用”白血病。

Itoh K et al.: "The role of soluble growth factors in inducing transient growth and clonal extinction of stromacell-dependenterythroblasticleukemiacells." Leukemia. 11. 1753-1761 (1997)

Itoh K 等人：“可溶性生长因子在诱导基质细胞依赖性红母细胞白血病细胞短暂生长和克隆消亡中的作用。”

Itoh K et al.: "Signalling mechanisms involved in long-term growth of ELM erythroleukaemiacells." Blood. 91. 1548-1555 (1998)

Itoh K 等人：“参与 ELM 红白血病细胞长期生长的信号机制。”