Endothelial cell-specific expression of tumor necrosis factor-α from the KDR or E-selectin promoters following retroviral delivery
Endothelial cell-specific expression of tumor necrosis factor-α from the KDR or E-selectin promoters following retroviral delivery
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DOI：
10.1089/hum.1997.8.18-2239
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发表时间：
1997-12-10
影响因子：
4.2
通讯作者：
Bicknell, R
中科院分区：
文献类型：
--
作者：
Jaggar, RT;Chan, HY;Bicknell, R
肿瘤血管系统为抗癌基因治疗提供了一个靶点，它具有良好的系统传递治疗的可及性和实体肿瘤类型之间的相对均质性。我们的目标是开发携带内皮特异性启动子的逆转录病毒，用于在体外将基因输送到增殖的内皮细胞和体内的肿瘤内皮细胞。本文报道了这种逆转录病毒载体的产生以及感染内皮细胞和间质成纤维细胞后小鼠肿瘤坏死因子-α(mtnf-α)的表达水平。携带mtnf-α的逆转录病毒载体的表达受逆转录病毒长末端重复序列或来自内皮特异性KDR/VEGE受体和E-选择素启动子的5‘端启动子控制。在自失活(SIN)载体骨架的背景下，KDR和E-选择素都被证明在肿瘤内皮细胞上表达上调，E-选择素启动子内的一个可能的多聚腺苷化序列AAATAAA被突变以允许携带该启动子的逆转录病毒载体的忠实传播。我们证明，与NIH-3T3成纤维细胞相比，内皮瘤细胞内启动子元件的mtnf-α表达增加了10-11倍。讨论了进一步改进载体设计的建议。
The tumor vasculature offers a target for anti-cancer gene therapy which has the advantages both of good accessibility to systemically delivered therapy and comparative homogeneity across solid tumor types, We aimed to develop retroviruses carrying endothelial-specific promoters for the delivery of genes to proliferating endothelial cells in vitro and to tumor endothelial cells in vivo, This paper reports the generation of such retroviral vectors and the level of expression of murine tumor necrosis factor-alpha (mTNF-alpha) cDNA following infection into endothelial cells and stromal fibroblasts, Retroviral vectors carrying mTNF-alpha have been generated whose expression is controlled either by the retroviral long terminal repeat or by 5' proximal promoter sequences from the endothelial-specific kinase insert domain receptor (KDR)/VEGE receptor and E-Selectin promoters within the context of a self-inactivating (SIN) vector backbone, Both KDR and E-Selectin have been shown to be upregulated on tumor endothelium, A putative polyadenylation sequence AAATAAA within the E-Selectin promoter was mutated to permit faithful transmission of retroviral vectors carrying this promoter, We demonstrate a 10- to 11-fold increase in mTNF-alpha expression from promoter elements within sEND endothelioma cells as compared to NIH-3T3 fibroblasts, Suggestions for further improvements in vector design are discussed.