Development of gene transfer by in vivo electroporation to the vascular wall

通过体内电穿孔向血管壁进行基因转移的进展

• 批准号: 14571126
• 负责人: MIYATA Tetsuro
• 金额: $ 2.18万
• 依托单位: The University of Tokyo
• 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-14571126/
• 关键词: electroporation; plasmid DNA; gene transfer

We developed a nonviral method for arterial gene transfer by in vivo electroporation with plasmid DNA. Electroporation is the physical process of including nanometreaized transient pores in a cell membrane by the application of short duration, high intensity electric field pulses to cells or tissues. The most important parameters for effective electroporation are: the stimulation voltage, the pulse duration, the electric current, plasmid concentration, and the shape of an electrode.Result: Electroporation with 30 voltage and pulse duration of 20 msec resulted in a marked increase of gene expression in vessel wall. The concentration of 400μ g/ml plasmid DNA is most optimal. The Bite of the vessel gene-transferred by in vivo electroporation is focused around the edge of the electrode. The same findings are demonstrated by the simulation model of vessel wall and the experiment of optical mapping. We compare with gene expression in vessel wall between with the monoplate electrode, which is one 5^*1mm plate, and the multiplate electrode, which is two plates in parallel. The gene expression using muitiplate electrode is superior to the one of monoplate electrode.In summary, we developed a method for gene transfer using electroporation, an approach which proved to the safe and more efficient than that using naked DNA. And then it will be possible that the site or the mount of gene expression is handled in the future.

我们开发了一种非病毒方法，用于用质粒DNA的体内电穿孔转移动脉基因。电穿孔是通过在细胞或组织中施加短持续时间，高强度的电场脉冲，将纳米成分的瞬态毛孔包括在细胞膜中。有效电穿孔的最重要参数是：刺激电压，脉冲持续时间，电流，质粒浓度以及电极的形状。解析：30电压和脉冲持续时间为20毫秒的电穿孔导致血管壁中基因表达的显着增加。 400μg/ml质粒DNA的浓度最佳。由体内电穿孔转移的血管基因的咬合集中在电极边缘周围。血管壁的仿真模型和光学映射的实验证明了相同的发现。我们与单板电极之间的血管壁中的基因表达进行比较，该电极是一个5^*1mm板，而乘电极则是两个并联的板。使用Mutiplate电极的基因表达优于单板电极之一。综上所述，我们使用电穿孔开发了一种基因转移的方法，这种方法比使用裸DNA的方法证明了这种方法对安全性，更有效的方法。然后，将来可能会处理基因表达的位点或基因表达。

项目成果

Inoue S, Koyama H, Miyata T, Shigematsu H: "Pathogenetic heterogeneity of in-stent lesion formation in human peripheral arterial disease."Journal of Vascular Surgery. 35. 672-678

Inoue S、Koyama H、Miyata T、Shigematsu H：“人类外周动脉疾病中支架内病变形成的致病异质性。”血管外科杂志。

Nishikage S, Koyama Y, Miyata T, Shigematsu H.: "In vivo electroporation enhances plasmid-based gene transfer of basic fibroblast growth factor for the treatment of ischemic limb."Journal of Surgical Research. (in press). (2004)

Nishikage S、Koyama Y、Miyata T、Shigematsu H.：“体内电穿孔增强碱性成纤维细胞生长因子的基于质粒的基因转移，用于治疗缺血性肢体。”外科研究杂志。

Miyata T, Sato O, Koyama H, Shigematsu H, Tada Y.: "Long-term survival after surgical treatment of patients with Takayasu's arteritis."Circulation. 108. 1474-1480 (2003)

Miyata T、Sato O、Koyama H、Shigematsu H、Tada Y.：“高安动脉炎患者手术治疗后的长期生存。”循环。

Miyata T, Koyama H: "Regenerative Medicine of Vascular"Respiratory and Circulatory. 50(4). 377-383 (2002)

Miyata T、Koyama H：“血管再生医学”呼吸和循环。

Inoue S, Koyama H, Miyata T, Shigematsu H: "Pathogenetic heterogeneity of in-stent lesion formation in human peripheral arterial disease."Journal of Vascular Surgery. 35. 672-678 (2002)

Inoue S、Koyama H、Miyata T、Shigematsu H：“人类外周动脉疾病中支架内病变形成的致病异质性。”血管外科杂志。