Analysis of cell-specific promoters for viral gene therapy targeted at the vascular endothelium
Analysis of cell-specific promoters for viral gene therapy targeted at the vascular endothelium
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DOI：
10.1161/01.hyp.38.1.65
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发表时间：
2001-07-01
期刊：
影响因子：
8.3
通讯作者：
Baker, AH
中科院分区：
文献类型：
--
作者：
Nicklin, SA;Reynolds, PN;Baker, AH
利用病毒载体进行针对内皮细胞的血管基因治疗受到载体的混杂性和病毒启动子的非特异性的限制，导致在多种组织中高水平的转基因表达。为了评估内皮细胞（ec）特异性启动子是否适合血管基因治疗，我们直接比较了鳍样酪氨酸激酶-1 （FLT-1）、细胞间粘附分子-2 （ICAM-2）和血管性血血病因子（vWF）启动子克隆到lacZ上游的腺病毒载体后驱动ec限制性转录的能力。观察到截然不同的表达谱。在体外培养的ECs中，FLT-1和ICAM-2启动子的转基因表达水平与巨细胞病毒相似，而vWF的表达水平极低。对非ec型的分析显示，ICAM-2而非FLT-1诱发了泄漏的转基因表达，从而确定FLT-1是最具选择性的启动子。在离体人基因治疗模型中，FLT-1启动子在完整的人静脉中表现出ec特异性的转基因表达，但在ec脱落的静脉中感染暴露的平滑肌细胞中没有检测到表达。此外，当腺病毒被全身注射到小鼠体内时，FLT-1启动子在肝脏（腺病毒在体内转导的主要靶器官）中表现出极低的基因表达。这项研究强调了FLT-1启动子在高血压及其并发症的局部和全身人类基因治疗中的潜力。
The use of viral vectors for vascular gene therapy targeted at the endothelium is limited by the promiscuous tropism of vectors and nonspecificity of viral promoters, resulting in high-level transgene expression in multiple tissues. To evaluate suitable endothelial cell (EC-specific promoters for vascular gene therapy, we directly compared the ability of the fins-like tyrosine kinase-1 (FLT-1), intercellular adhesion molecule-2 (ICAM-2), and von Willebrand factor (vWF) promoters to drive EC-restricted transcription after cloning into adenoviral vectors upstream of lacZ. Vastly different expression profiles were observed. Whereas both FLT-1 and ICAM-2 promoters generated transgene expression levels similar to cytomegalovirus in ECs in vitro, vWF expression levels were extremely low. Analysis of non-EC types revealed that ICAM-2 but not FLT-1 evoked leaky transgene expression, thus identifying FLT-1 as the most selective promoter. With an ex vivo human gene therapy model, the FLT-1 promoter demonstrated EC-specific transgene expression in intact human vein but no detectable expression from infected exposed smooth muscle cells in EC-denuded vein. Furthermore, when adenoviruses were systemically administered to mice, the FLT-1 promoter demonstrated extremely low-level gene expression in the liver, the major target organ for adenoviral transduction in vivo. This study highlights the potential of using the FLT-1 promoter for local and systemic human gene therapy in hypertension and its complications.