Development of efficient plasmid DNA transfer into adult rat central nervous system using microbubble-enhanced ultrasound
Development of efficient plasmid DNA transfer into adult rat central nervous system using microbubble-enhanced ultrasound
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DOI：
10.1038/sj.gt.3302323
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发表时间：
2004-10-01
期刊：
影响因子：
5.1
通讯作者：
Morishita, R
中科院分区：
文献类型：
--
作者：
Shimamura, M;Sato, N;Morishita, R
虽然基因治疗可能成为治疗中枢神经系统疾病的一种有前途的方法，但安全性问题是人类基因治疗中需要认真考虑的问题。为了克服这个问题，我们开发了一种基于质粒DNA、使用微泡增强超声方法将基因转移到成年大鼠大脑中的有效方法，因为微泡增强超声已显示出将基因转染到血管等其他组织中的前景。使用微泡增强超声方法，荧光素酶表达增加约10倍，相比单独注射裸质粒DNA。有趣的是，与以前使用病毒的研究相反，基因表达的位点仅限于脑池内注射的声波照射位点。在中枢神经系统中很容易检测到报告基因Venus的表达。脑池内注射主要在脑膜细胞中检测到转染细胞，纹状体内注射主要在胶质细胞中检测到转染细胞。微泡增强超声没有明显的组织损伤证据。总体而言，本研究证明了质粒DNA有效转移到中枢神经系统的可行性，为治疗肿瘤等各种疾病提供了新的选择。
Although gene therapy might become a promising approach for central nervous system diseases, the safety issue is a serious consideration in human gene therapy. To overcome this problem, we developed an efficient gene transfer method into the adult rat brain based on plasmid DNA using a microbubble-enhanced ultrasound method, since microbubble-enhanced ultrasound has shown promise for transfecting genes into other tissues such as blood vessels. Using the microbubble-enhanced ultrasound method, luciferase expression was increased approximately 10-fold as compared to injection of naked plasmid DNA alone. Interestingly, the site of gene expression was limited to the site of insonation with intracisternal injection, in contrast to previous studies using viruses. Expression of the reporter gene, Venus, was readily detected in the central nervous system. The transfected cells were mainly detected in meningeal cells with intracisternal injection, and in glial cells with intrastriatal injection. There was no obvious evidence of tissue damage by microbubble-enhanced ultrasound. Overall, the present study demonstrated the feasibility of efficient plasmid DNA transfer into the central nervous system, providing a new option for treating various diseases such as tumors.