Development of novel gene delivery systems based on the function of biological molecules

基于生物分子功能的新型基因传递系统的开发

• 批准号: 10557234
• 负责人: YUTSUDO Masuo
• 金额: $ 8万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10557234/
• 关键词: DNA; Nuclear targeting; Phage; Telomere; Cancer; Sendai virus; Fusogenic liposome; Gene therapy; TRF1; 染色体; ハイブリッドベクター; ガン胎児抗原; ナトリウムチャンネル; ラムダファージ; 細胞膜; 核膜; Protein Transduction Domain; 人工染色体; RNAレプリコン; 遺伝子導入; ベクター

In this research project, we investigate the essential function of biological systems, such as viruses and chromosomes, which will be useful for developing novel non-viral devices for gene transfer and expression. The results we have obtained during the term of this project are summarized as below.1. We analyzed the mechanism of membrane fusion that will mediated efficient DNA delivery, and found that the membrane fusion requires essential non-receptor molecules which may be interacted with HN protein of Sendai virus.2. We establish the system to display various biological peptides on the surface of lambda phage particles, and propose to use this system to analyze the function of these peptides as components of non-viral gene delivery vehicles. Using this system, we showed that Tat peptide could enhance the delivery of lambda phage DNA into the cells and that NLS signal peptide of SV40 T antigen could target the lambda phage particles into the nucleus through the nuclear pore complex.3. We analyze the mechanism by which human chromosomes are stabilized in the nucleus, and found that the interaction between TRF1 and telomere repeat was involved in this process.4. We succeeded to cure mice with the disseminated cancer by treating them with the fusogenic liposomes encapsulating cancer-specific suicide gene.

在本研究项目中，我们研究了生物系统的基本功能，如病毒和染色体，这将有助于开发用于基因转移和表达的新型非病毒装置。我们在本项目期间所取得的成果总结如下.我们分析了膜融合介导DNA高效转运的机制，发现膜融合需要与仙台病毒HN蛋白相互作用的非受体分子.我们建立了在λ噬菌体颗粒表面展示各种生物肽的系统，并建议使用该系统来分析这些肽作为非病毒基因递送载体的组分的功能。利用该系统，我们发现达特肽可以增强λ噬菌体DNA进入细胞的递送，并且SV 40 T抗原的NLS信号肽可以通过核孔复合物将λ噬菌体颗粒靶向细胞核.我们分析了人类染色体稳定在细胞核中的机制，发现TRF1和端粒重复序列之间的相互作用参与了这一过程.我们成功地治愈了小鼠与扩散的癌症治疗与融合脂质体包裹癌症特异性自杀基因。

项目成果

M.Shimakage, et al.: "Expression of Epstein-Barr virus in cutaneous T-cell lymphoma including mycosis fungoides"International Journal of Cancer. 92. 226-231 (2001)

M.Shimakage 等人：“Epstein-Barr 病毒在皮肤 T 细胞淋巴瘤（包括蕈样肉芽肿）中的表达”国际癌症杂志。

中西真人, 他: "遺伝子治療ハンドブック"株式会社エヌ・ティー・エス. 1085 (1999)

Masato Nakanishi 等人：《基因治疗手册》NTS Co., Ltd. 1085 (1999)

Miyake,K.,et al.: "Transforming growth factor-beta1 stimulates contraction of human glioblastoma cell-mediated collagen lattice through enhanced alpha 2 integrin expression" Journal of Experimental Therapeutics and Oncology. (印刷中).

Miyake, K. 等人：“转化生长因子-β1 通过增强 α2 整合素表达刺激人胶质母细胞瘤细胞介导的胶原蛋白晶格收缩”《实验治疗与肿瘤学杂志》（正在出版）。

Nakanishi,M.,et al.: "Gene Delivery Systems Using the Sendai Virus" Molecular Membrane Biology. (印刷中).

Nakanishi, M. 等人：“使用仙台病毒的基因传递系统”分子膜生物学（正在出版）。

M.Nakanishi, et al.: "Gene Delivery Systems Using the Sendai Virus"Mol. Memb. Biol.. 16. 123-127 (1999)

M.Nakanishi 等人：“使用仙台病毒的基因传递系统”Mol。