Dzau V J, Mann M J, Morishita R, Kaneda Y
Research Institute, Brigham and Women's Hospital, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11421-5. doi: 10.1073/pnas.93.21.11421.
To improve the efficiency of liposome-mediated DNA transfer as a tool for gene therapy, we have developed a fusigenic liposome vector based on principles of viral cell fusion. The fusion proteins of hemagglutinating virus of Japan (HVJ; also Sendai virus) are complexed with liposomes that encapsulate oligodeoxynucleotide or plasmid DNA. Subsequent fusion of HVJ-liposomes with plasma membranes introduces the DNA directly into the cytoplasm. In addition, a DNA-binding nuclear protein is incorporated into the HVJ-liposome particle to enhance plasmid transgene expression. The fusigenic viral liposome vector has proven to be efficient for the intracellular introduction of oligodeoxynucleotide, as well as intact genes up to 100 kbp, both in vitro and in vivo. Many animal tissues have been found to be suitable targets for fusigenic viral liposome DNA transfer. In the cardiovascular system, we have documented successful cytostatic gene therapy in models of vascular proliferative disease using antisense oligodeoxynucleotides against cell cycle genes, double-stranded oligodeoxynucleotides as "decoys" to trap the transcription factor E2F, and expression of a transgene encoding the constitutive endothelial cell form of nitric oxide synthase. Similar strategies are also effective for the genetic engineering of vein grafts and for the treatment of a mouse model of immune-mediated glomerular disease.
为提高脂质体介导的DNA转移作为基因治疗工具的效率，我们基于病毒细胞融合原理开发了一种融合脂质体载体。日本血凝病毒（HVJ；也称为仙台病毒）的融合蛋白与包裹寡脱氧核苷酸或质粒DNA的脂质体复合。随后，HVJ-脂质体与质膜融合，将DNA直接导入细胞质。此外，一种与DNA结合的核蛋白被整合到HVJ-脂质体颗粒中，以增强质粒转基因的表达。融合病毒脂质体载体已被证明在体外和体内对寡脱氧核苷酸以及长达100kbp的完整基因的细胞内导入都是有效的。已发现许多动物组织是融合病毒脂质体DNA转移的合适靶标。在心血管系统中，我们已证明在血管增殖性疾病模型中使用针对细胞周期基因的反义寡脱氧核苷酸、作为“诱饵”捕获转录因子E2F的双链寡脱氧核苷酸以及编码组成型内皮细胞形式一氧化氮合酶的转基因表达进行的成功的细胞生长抑制基因治疗。类似的策略对静脉移植物的基因工程以及免疫介导的肾小球疾病小鼠模型的治疗也有效。
