Shimamura Munehisa, Morishita Ryuichi, Endoh Masayuki, Oshima Kazuo, Aoki Motokuni, Waguri Satoshi, Uchiyama Yasuo, Kaneda Yasufumi
Division of Gene Therapy Science, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita 565-0871, Osaka, Japan.
Biochem Biophys Res Commun. 2003 Jan 10;300(2):464-71. doi: 10.1016/s0006-291x(02)02807-3.
To overcome some problems of virus vectors, we developed a novel non-viral vector system, the HVJ-envelope vector (HVJ-E). In this study, we investigated the feasibility of gene transfer into the CNS using the HVJ-E both in vitro and in vivo. Using the Venus reporter gene, fluorescence could be detected in cultured rat cerebral cortex neurons and glial cells. In vivo, the reporter gene (Venus) was successfully transfected into the rat brain by direct injection into the thalamus, intraventricular injection, or intrathecal injection, without inducing immunological change. When the vector was injected after transient occlusion of the middle cerebral artery, fluorescence due to EGFP gene or luciferase activity could be detected only in the injured hemisphere. Finally, luciferase activity was markedly enhanced by the addition of 50 U/ml heparin (P<0.01). Development of efficient HVJ-E for gene transfer into the CNS will be useful for research and clinical gene therapy.
为克服病毒载体的一些问题，我们开发了一种新型非病毒载体系统，即HVJ包膜载体（HVJ-E）。在本研究中，我们研究了使用HVJ-E在体外和体内将基因导入中枢神经系统的可行性。使用金星报告基因，可在培养的大鼠大脑皮质神经元和神经胶质细胞中检测到荧光。在体内，通过直接注入丘脑、脑室内注射或鞘内注射，报告基因（金星）成功转染到大鼠大脑中，且未引起免疫变化。当中脑动脉短暂闭塞后注射载体时，仅在受损半球可检测到由于增强绿色荧光蛋白（EGFP）基因或荧光素酶活性产生的荧光。最后，添加50 U/ml肝素可使荧光素酶活性显著增强（P<0.01）。开发用于将基因导入中枢神经系统的高效HVJ-E将有助于研究和临床基因治疗。
