In vivo targeting of tumor endothelial cells by systemic delivery of lentiviral vectors
In vivo targeting of tumor endothelial cells by systemic delivery of lentiviral vectors
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DOI：
10.1089/104303403322168028
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发表时间：
2003-08-01
影响因子：
4.2
通讯作者：
Naldini, L
中科院分区：
文献类型：
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作者：
De Palma, M;Venneri, MA;Naldini, L
肿瘤血管生成是肿瘤生长的限速因子，并且肿瘤血管的内皮细胞显示出可用于选择性递送癌症治疗剂的特定特征。为了特异性靶向外源基因的血管生成肿瘤血管，我们产生了一组水泡性口炎病毒假型慢病毒载体（LV）的内皮细胞（EC）特异性表达工程。我们克隆了广泛的转录调控序列从基因优先表达的内皮细胞（Tie 1，Tie 2，Flk-1，VE-Cad，和ICAM-2）到自我失活的LV驱动的标志物基因编码绿色荧光蛋白（GFP）或条件毒性基因编码硝基还原酶的表达，并将它们与无处不在的表达磷酸甘油酸激酶（PGK）和巨细胞病毒（CMV）启动子。我们评估了一组人原代培养物（包括EC、成纤维细胞、神经元、淋巴细胞和造血祖细胞）和肿瘤细胞系中载体表达的体外效率和特异性。我们发现含有Tie 2基因的启动子和增强子序列的载体在体外和体内的EC中实现了显着的表达特异性。在静脉内递送到荷瘤小鼠中时，Tie 2载体靶向表达至肿瘤血管的EC。相反，携带PGK或CMV启动子的LV在肿瘤和其他器官的EC和非EC中产生广泛的GFP标记。先前报道的上调Tie 2基因在血管生成激活的EC中可以解释Tie 2载体在肿瘤血管EC中表达的显著选择性。这种新的载体为选择性地将基因治疗递送到体内肿瘤部位提供了手段。
Tumor angiogenesis is a rate-limiting factor for tumor growth, and the endothelial cells of tumor vessels display specific features that can be exploited for the selective delivery of cancer therapeutics. To specifically target exogenous genes to angiogenic tumor vessels, we generated a panel of vesicular stomatitis virus-pseudo-typed lentiviral vectors (LVs) engineered for endothelial cell (EC)-specific expression. We cloned a wide repertoire of transcription regulatory sequences from genes preferentially expressed in ECs (Tie1, Tie2, Flk-1, VE-Cad, and ICAM-2) into self-inactivating LVs to drive expression of the marker gene encoding green fluorescent protein (GFP) or of the conditionally toxic gene encoding nitroreductase, and compared them with the ubiquitously expressing phosphoglycerate kinase (PGK) and cytomegalovirus (CMV) promoters. We evaluated the efficiency and specificity of vector expression in vitro in a panel of human primary cultures, including ECs, fibroblasts, neurons, lymphocytes, and hematopoietic progenitors, and in tumor cell lines. We found that vectors containing promoter and enhancer sequences from the Tie2 gene achieved remarkable specificity of expression in ECs in vitro and in vivo. On intravenous delivery into tumor-bearing mice, the Tie2 vector targeted expression to the ECs of tumor vessels. In contrast, LVs carrying the PGK or CMV promoter gave widespread GFP marking in ECs and non-ECs of tumors and other organs. The previously reported upregulation of the Tie2 gene in ECs activated for angiogenesis may explain the remarkable selectivity of expression of the Tie2 vector in ECs of tumor vessels. The new vector provides the means for selective delivery of gene therapy to tumor sites in vivo.