Liver-targeted gene therapy by SV40-based vectors using the hydrodynamic injection method
Liver-targeted gene therapy by SV40-based vectors using the hydrodynamic injection method
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DOI：
10.1089/hum.2005.16.361
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发表时间：
2005-03-01
影响因子：
4.2
通讯作者：
Oppenheim, A
中科院分区：
文献类型：
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作者：
Arad, U;Zeira, E;Oppenheim, A
肝细胞中缺陷基因的有效重建可用于通过基因治疗治疗各种肝脏和全身性疾病。为了探索基于SV 40的载体在肝脏基因治疗中的潜力，我们构建了SV/luc，一种SV 40 T抗原替代转导载体，其在COS和COT细胞上增殖，COS和COT细胞以反式提供SV 40 T抗原。通过尾静脉向BALB/C小鼠注射体积为1-2 ml的含有3 × 10（6）至10（8）个转导单位的SV/luc储备液。荧光素酶活性用光检测冷却电荷耦合器件（CCCD）相机监测，其使得能够连续体内测量luc表达。SV 40载体被证明是有效的基因传递到肝脏，导致长期（>= 107天）的转基因表达在肝细胞。用3 × 10（6）至3 × 10（7）个转导单位获得最佳结果。流体动力学载体递送方法引起短暂的肝脏炎症变化，在几天内完全恢复。在治疗小鼠的血清中检测到低水平的SV 40中和抗体;然而，没有迹象表明载体或转基因特异性细胞免疫应答。使用重组衣壳蛋白和质粒DNA在体外包装的载体在肝转导中也有效。这些结果表明，SV 40载体可能是有用的肝脏基因治疗。
Efficient reconstitution of defective genes in hepatocytes could be used to treat various liver and systemic diseases through gene therapy. To explore the potential of SV40-based vectors in liver gene therapy, we constructed SV/luc, an SV40 T-antigen replacement transduction vector, that was propagated on COS and COT cells, which supply the SV40 T-antigen in trans. For liver targeting, BALB/C mice were injected via the tail vein with SV/luc stocks containing 3 x 10(6) to 10(8) transducing units in a volume of 1-2 ml. Luciferase activity was monitored with a light-detection cooled charged-coupled device (CCCD) camera, which enables continuous in vivo measurement of luc expression. The SV40 vector proved to be efficient in gene delivery to the liver, leading to long-term (>= 107 days) transgene expression in hepatocytes. Optimal results were obtained with 3 x 10(6) to 3 x 10(7) transducing units. The hydrodynamic vector delivery method caused transient liver inflammatory changes, with full recovery within days. Low levels of SV40- neutralizing antibodies were detected in the sera of treated mice; however, there was no indication of vector or transgene-specific cellular immune responses. Vectors packaged in vitro, using recombinant capsid proteins and plasmid DNA, were also effective in liver transduction. These results suggest that SV40 vectors may be useful for liver gene therapy.