Development of gene therapy using a novel domestic viral vector.

使用新型国产病毒载体开发基因治疗。

• 批准号: 14570752
• 负责人: KUSUHARA Koichi
• 金额: $ 2.56万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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/en/grant/KAKENHI-PROJECT-14570752/
• 关键词: gene therapy; hematopoietic stem cells; gene transfer; Sendai virus; vector; cord blood

Hematopoietic stem cells (HSCs) are a promising target for gene therapy, however, the low efficiencies of gene transfer using currently available vectors face practical limitations. We have recently developed a novel and efficient gene transfer agent, namely recombinant Sendai virus (SeV). We characterized SeV-mediated gene transfer to human cord blood (CB) HSCs and primitive progenitor cells (PPC) using the jelly fish green fluorescent protein (GFP) gene. Even at a relatively low titer, SeV achieved highly efficient GFP expression in GB CD34^+ cells, as well as more immature CB progenitor cells, CD34^+AC133^+ and GD34^+CD38^-cells, without cytokines prestimulation, that was a clear contrast to the features of gene, transfer using retroviruses. SeV-mediated gene transfer was not seriously affected by the cell cycle status : GFP was expressed in significant percentage of the G_0 cells. In vitro cell differentiation studies revealed that gene transfer occurred in progenitor cells of all lineages (GM-CFU, BFU-E, Mix-CFUand Total). These findings showed that SeV could prove to be a promising vector for efficient gene transfer to CB HSCs, while preserving their ability to reconstitute the entire hematopoietic series. in vivo gene transfer experiments using SeV with temperature-sensitive mutation are now underway.

造血干细胞（HSC）是基因治疗的一个有前途的靶标，然而，使用现有载体的基因转移效率低，面临实际限制。我们最近开发了一种新型高效的基因转移剂，即重组仙台病毒（SeV）。我们使用水母绿色荧光蛋白 (GFP) 基因表征了 SeV 介导的人脐带血 (CB) HSC 和原始祖细胞 (PPC) 基因转移。即使在相对较低的滴度下，SeV 在 GB CD34^+ 细胞以及更不成熟的 CB 祖细胞 CD34^+AC133^+ 和 GD34^+CD38^-细胞中也实现了高效的 GFP 表达，无需细胞因子预刺激，这与使用逆转录病毒进行基因转移的特征形成鲜明对比。 SeV介导的基因转移并未受到细胞周期状态的严重影响：GFP在G_0细胞的显着百分比中表达。体外细胞分化研究表明，所有谱系（GM-CFU、BFU-E、Mix-CFU 和 Total）的祖细胞中都发生了基因转移。这些发现表明，SeV 可能被证明是一种有前途的载体，可将基因有效转移至 CB HSC，同时保留其重建整个造血系列的能力。使用带有温度敏感突变的 SeV 进行体内基因转移实验正在进行中。

项目成果

Ikeda Y, Yonemitsu Y et al.: "Recombinant Sendai virus-mediated gene transfer into adult rat retinal tissue : efficient gene transfer by brief exposure."Exp Eye Res. 75(1). 39-48 (2002)

Ikeda Y、Yonemitsu Y 等人：“重组仙台病毒介导的基因转移到成年大鼠视网膜组织中：通过短暂暴露进行有效的基因转移。”Exp Eye Res。

Jin CH, Kusuhara K et al.: "Recombinant Sendai virus provides a highly efficient gene transfer into human cord blood-derived hematopoietic stem cells."Gene Ther. 10(3). 272-277 (2003)

Jin CH、Kusuhara K 等人：“重组仙台病毒可高效地将基因转移到人脐带血来源的造血干细胞中。”Gene Ther。

Shoji T, Yonemitsu Y, et al.: "Intramuscular gene transfer of FGF-2 attenuates endothelial dysfunction and inhibits intimal hyperplasia of vein grafts in poor-runoff limbs of rabbit."Am J Physiol Heart Circ Physiol. 285(1). H173-H182 (2003)

Shoji T、Yonemitsu Y 等人：“FGF-2 的肌肉内基因转移可减轻内皮功能障碍，并抑制兔径流不良四肢静脉移植物的内膜增生。”Am J Physiol Heart Circ Physiol。

Kusuhara K, Nomura A et al.: "Tumour necrosis factor receptor-associated periodic syndrome with a novel mutation in the TNFRSF1A gene in a Japanese family."Eur J Pediatr. 163(1). 30-32 (2004)

Kusuhara K、Nomura A 等人：“日本家族中具有 TNFRSF1A 基因新突变的肿瘤坏死因子受体相关周期性综合征。”Eur J Pediatr。

Okano S, Yonemitsu Y, et al.: "Recombinant Sendai virus vectors for activated T lymphocytes."Gene Ther. 10(16). 1381-1391 (2003)

Okano S、Yonemitsu Y 等人：“用于激活 T 淋巴细胞的重组仙台病毒载体。”Gene Ther。