Morishita Norio, Nakagami Hironori, Morishita Ryuichi, Takeda Shin-ichi, Mishima Fumihito, Terazono Bungo, Nishijima Shigehiro, Kaneda Yasufumi, Tanaka Noriaki
Department of Gastroenterological Surgery, Transplant, and Surgical Oncology, Graduate School of Medicine and Dentistry, Okayama University, Okayama, Japan.
Biochem Biophys Res Commun. 2005 Sep 9;334(4):1121-6. doi: 10.1016/j.bbrc.2005.06.204.
To enter the realm of human gene therapy, a novel drug delivery system is required for efficient delivery of small molecules with high safety for clinical usage. We have developed a unique vector "HVJ-E (hemagglutinating virus of Japan-envelope)" that can rapidly transfer plasmid DNA, oligonucleotide, and protein into cells by cell-fusion. In this study, we associated HVJ-E with magnetic nanoparticles, which can potentially enhance its transfection efficiency in the presence of a magnetic force. Magnetic nanoparticles, such as maghemite, with an average size of 29 nm, can be regulated by a magnetic force and basically consist of oxidized Fe which is commonly used as a supplement for the treatment of anemia. A mixture of magnetite particles with protamine sulfate, which gives a cationic surface charge on the maghemite particles, significantly enhanced the transfection efficiency in an in vitro cell culture system based on HVJ-E technology, resulting in a reduction in the required titer of HVJ. Addition of magnetic nanoparticles would enhance the association of HVJ-E with the cell membrane with a magnetic force. However, maghemite particles surface-coated with heparin, but not protamine sulfate, enhanced the transfection efficiency in the analysis of direct injection into the mouse liver in an in vivo model. The size and surface chemistry of magnetic particles could be tailored accordingly to meet specific demands of physical and biological characteristics. Overall, magnetic nanoparticles with different surface modifications can enhance HVJ-E-based gene transfer by modification of the size or charge, which could potentially help to overcome fundamental limitations to gene therapy in vivo.
为了进入人类基因治疗领域，需要一种新型药物递送系统，以便高效递送小分子且具有临床使用的高安全性。我们开发了一种独特的载体“HVJ-E（日本血凝病毒包膜）”，它可以通过细胞融合将质粒DNA、寡核苷酸和蛋白质快速转移到细胞中。在本研究中，我们将HVJ-E与磁性纳米颗粒相结合，在磁力存在的情况下，磁性纳米颗粒可能会提高其转染效率。平均粒径为29 nm的磁性纳米颗粒，如磁赤铁矿，可以通过磁力进行调控，其主要由氧化铁组成，氧化铁通常用作治疗贫血的补充剂。磁铁矿颗粒与硫酸鱼精蛋白的混合物，可使磁赤铁矿颗粒表面带阳离子电荷，在基于HVJ-E技术的体外细胞培养系统中显著提高了转染效率，从而降低了所需的HVJ滴度。添加磁性纳米颗粒会通过磁力增强HVJ-E与细胞膜的结合。然而，在体内模型中对小鼠肝脏进行直接注射分析时，表面包被肝素而非硫酸鱼精蛋白的磁赤铁矿颗粒提高了转染效率。可以相应地调整磁性颗粒的大小和表面化学性质，以满足物理和生物学特性的特定要求。总体而言，具有不同表面修饰的磁性纳米颗粒可以通过改变大小或电荷来增强基于HVJ-E的基因转移，这可能有助于克服体内基因治疗的基本限制。
