Improvement of muscle function by introducing genes by electroporation following intramuscular injection

肌肉注射后通过电穿孔导入基因改善肌肉功能

基本信息

批准号：
13670063
负责人：
MIYAZAKI Jun-ichi
金额：
$ 0.77万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670063/
关键词：
electroporation gene therapy gene transfer expression vector Intramuscular injection

项目摘要

The proportion of aged people in the whole population is increasing rapidly, so that maintaining skeletal muscle function is becoming more important for the healthy life of people. Recently, various kinds of methods for in vivo gene transfer have been developed. For maintaining muscle function, direct plasmid DNA injection into muscle might be useful for improvement of muscle function. Previously, I demonstrated that gene transfer into muscle by electroporation in vivo was far more efficient than simple intramuscular DNA injection. In this study, I speculated that this method might be used to allow rat muscle to express some genes related to muscle function, insulin-like growth factor 1 (IGF-1) for proliferation of muscle cells, vascular endothelial growth factor (VEGF) for angiogenesis, and growth hormone(GH). And some indices, concentration of DNA, DNA solution, structure of electrodes and frequency of pulse, were analyzed for improvement of efficiency of this method. Vectors inserted with CDNA of these genes were introduced into cultured cells, showing that products of these genes were secreted into the culture media. As a control study, a plasmid containing erythropoietin cDNA was introduced into rat muscle. The results showed that the best condition for gene transfer was as follows: injection of 400 μg of DNA and electroporation with 5mm-electrode needle, 8 times of 50 msec-pulse at 100V.Intramuscular injection following by electroporation in vivo is simple and efficient for gene transfer, and may provide a potential approach toward systemic delivery of cytokines , growth factors, and other serum proteins for human gene therapy. More improvement of this method could contribute for human hearth.

整个人群中老年人的比例正在迅速增加，因此维持骨骼肌肉功能对人的健康生活变得越来越重要。最近，已经开发了各种体内基因转移的方法。为了维持肌肉功能，直接质粒DNA注射到肌肉中可能有助于改善肌肉功能。以前，我证明了通过体内电穿孔转移到肌肉的基因比简单的肌内DNA注射效率要高得多。在这项研究中，我推测该方法可以用来使大鼠肌肉表达与肌肉功能，胰岛素样生长因子1（IGF-1）相关的一些基因，用于肌肉细胞的增殖，血管内皮生长因子（VEGF）用于血管生成和生长（GH）。分析了一些指数，DNA溶液的浓度，电子结构和脉冲频率，以提高该方法的效率。将这些基因的cDNA插入的载体引入培养细胞中，表明这些基因的产物分泌到培养基中。作为对照研究，将含有红细胞生成素cDNA的质粒引入大鼠肌肉中。结果表明，基因转移的最佳条件如下：注射400μg的DNA和用5mm-电极针的电穿孔，在100V时在100V时进行8次50毫秒 - 脉冲。通过体内电穿孔后，intramuscular注入了基因转移的简单和有效的基因转移，并且可以提供cytokines of Sertokine gene Gene，以及其他人类的生长因子，并提供其他疗法。这种方法的更多改进可能有助于人类壁炉。