Sindbis Virus-Pseudotyped Lentiviral Vectors Carrying VEGFR2-Specific Nanobody for Potential Transductional Targeting of Tumor Vasculature
Sindbis Virus-Pseudotyped Lentiviral Vectors Carrying VEGFR2-Specific Nanobody for Potential Transductional Targeting of Tumor Vasculature
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DOI：
10.1007/s12033-016-9973-7
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发表时间：
2016-11-01
影响因子：
2.6
通讯作者：
Azadmanesh, Kayhan
中科院分区：
文献类型：
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作者：
Ahani, Roshank;Roohvand, Farzin;Azadmanesh, Kayhan
将选择性/特异性引入以病毒为基础的基因递送系统，如慢病毒载体，在它们用于癌症基因治疗的系统给药中至关重要。肿瘤相关血管内皮细胞(TAECs)在肿瘤血管生成中的关键作用和血管内皮生长因子受体2(VEGFR2或KDR)在TAECs中的高表达使其成为肿瘤治疗的有效靶点。在此，我们报道了VEGFR2靶向与Sindbis病毒E2糖蛋白嵌合的假型LV(CSVE2s)的发展。为此，将VEGFR2特异性纳米体的序列或其天然配体(VEGF(121))插入到Sindbis病毒E2糖蛋白的结合位置。在电子计算机模拟中，数据表明插入的靶向基序是在cSVE2的背景下暴露的。对LV的Western印迹分析表明，cSVE2被掺入病毒颗粒。捕获酶联免疫吸附试验证实了所掺入的cSVE2的特异性/功能性。用构建的LV转导293/KDR(表达VEGFR2)或293T细胞(阴性对照)，荧光显微镜和流式细胞仪分析表明，这两个靶向基序选择性转导293/KDR细胞(30%)，而293T对照细胞(1-2%)。这些结果表明，与血管内皮生长因子(121)相比，VEGFR2特异性纳米体具有相似的靶向特性，并表明显示LV的纳米体具有转导靶向肿瘤血管的潜力。
Introduction of selectivity/specificity into viral-based gene delivery systems, such as lentiviral vectors (LVs), is crucial in their systemic administration for cancer gene therapy. The pivotal role of tumor-associated endothelial cells (TAECs) in tumor angiogenesis and overexpression of vascular endothelial growth factor receptor-2 (VEGFR2 or KDR) in TAECs makes them a potent target in cancer treatment. Herein, we report the development of VEGFR2-targeted LVs pseudotyped with chimeric sindbis virus E2 glycoprotein (cSVE2s). For this purpose, either sequence of a VEGFR2-specific nanobody or its natural ligand (VEGF(121)) was inserted into the binding site of sindbis virus E2 glycoprotein. In silico modeling data suggested that the inserted targeting motifs were exposed in the context of cSVE2s. Western blot analysis of LVs indicated the incorporation of cSVE2s into viral particles. Capture ELISA demonstrated the specificity/functionality of the incorporated cSVE2s. Transduction of 293/KDR (expressing VEGFR2) or 293T cells (negative control) by constructed LVs followed by fluorescent microscopy and flow cytometric analyses indicated selective transduction of 293/KDR cells (30 %) by both targeting motifs compared to 293T control cells (1-2 %). These results implied similar targeting properties of VEGFR2-specific nanobody compared to the VEGF(121) and indicated the potential for transductional targeting of tumor vasculature by the nanobody displaying LVs.