Transposon-based interferon gamma gene transfer overcomes limitations of episomal plasmid for immunogene therapy of glioblastoma.
Transposon-based interferon gamma gene transfer overcomes limitations of episomal plasmid for immunogene therapy of glioblastoma.
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基于转座子的干扰素γ基因转移克服了附加型质粒在胶质母细胞瘤免疫基因治疗中的局限性。
DOI：
10.1038/sj.cgt.7701045
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发表时间：
2007
影响因子：
6.4
通讯作者：
Ohlfest,JR
中科院分区：
文献类型：
--
作者：
Wu,A;Oh,S;Ericson,K;Demorest,ZL;Vengco,I;Gharagozlou,S;Chen,W;Low,WC;Ohlfest,JR
尽管基因传递技术有所改进，但质粒 DNA 的瞬时表达限制了非病毒载体应用于癌症基因治疗的功效。我们之前利用睡美人 (SB) 转座元件开发了能够在人胶质母细胞瘤细胞中进行转基因整合和长期表达的质粒 DNA 载体。在这项研究中，我们在同基因 GL261 神经胶质瘤模型中比较了使用附加型或 SB 载体的干扰素 γ (IFN-γ) 免疫基因治疗的疗效。基因递送是通过肿瘤内对流增强 DNA/聚乙烯亚胺复合物的递送来实现的。只有用编码 SB 转座酶的 DNA 处理以促进染色体整合的小鼠才表现出存活率的显着增加（P < 0.05）。通过逆转录聚合酶链式反应评估，SB介导的瘤内基因转移引起载体衍生的和内源性IFN-γ的持续IFN-γ表达，而附加型质粒基因转移后的表达在2周内检测不到。当 SB 介导的 IFN-γ 基因转移与 CpG 寡脱氧核苷酸联合作为辅助治疗时，中位生存期进一步提高。生存期延长与体内生物发光成像测量的肿瘤消退以及 ELISPOT 测定显示的 T 细胞活化增强呈正相关。 SB 似乎通过延长转基因表达的持续时间来提高使用非病毒载体的细胞因子基因治疗的功效。
Despite improvements in gene delivery technology, transient expression of plasmid DNA has limited the efficacy of nonviral vectors applied to cancer gene therapy. We previously developed plasmid DNA vectors capable of transgene integration and long-term expression in human glioblastoma cells by utilizing the Sleeping Beauty (SB) transposable element. In this study, we compared the efficacy of interferon gamma (IFN-γ) immunogene therapy using episomal or SB vectors in a syngeneic GL261 glioma model. Gene delivery was achieved by intratumoral convection-enhanced delivery of DNA/polyethylenimine complexes. Only mice treated with SB transposase-encoding DNA to facilitate chromosomal integration exhibited a significant increase in survival (P< 0.05). SB-mediated intratumoral gene transfer caused sustained IFN-γ expression assessed by reverse transcription-polymerase chain reaction, of both vector-derived and endogenous IFN-γ, whereas expression following episomal plasmid gene transfer was undetectable within 2 weeks. Median survival was enhanced further when SB-mediated IFN-γ gene transfer was combined with CpG oligodeoxynucleotides as adjuvant therapy. Prolonged survival positively correlated with tumor regression measured by in vivo bioluminescent imaging, and enhanced T-cell activation revealed by the ELISPOT assay. SB appears to improve the efficacy of cytokine gene therapy using nonviral vectors by enhancing the duration of transgene expression.