Gene delivery to in situ veins: Differential effects of adenovirus and adeno-associated viral vectors
Gene delivery to in situ veins: Differential effects of adenovirus and adeno-associated viral vectors
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DOI：
10.1016/s0741-5214(00)90105-8
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发表时间：
2000-06-01
影响因子：
4.3
通讯作者：
Conte, MS
中科院分区：
文献类型：
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作者：
Eslami, MH;Gangadharan, SP;Conte, MS
目的：基因转移提供了在外科植入时改变静脉移植物生物学的可能性。基因递送的效率、表达的稳定性和宿主反应是候选载体的关键参数。我们比较了腺病毒（AD）和腺相关病毒（AAV）载体对转基因表达和单核细胞粘附（MA）在治疗vein segments.Methods：成年新西兰白色兔（N = 51）的影响腔内暴露麻醉，颈静脉插管双侧。用载体（2.10（8）至1.10（10）感染颗粒/mL）或媒介物（对照）轻轻扩张静脉30分钟，之后恢复静脉血流。使用编码标记基因β-半乳糖苷酶（LacZ）和绿色荧光蛋白（GFP）的AD和AAV载体。感染后2 - 40天，将血管取出，进行基因表达分析（X-gal染色，逆转录-聚合酶链反应），MA和免疫组化。离体粘附测定使用Cr-51标记的THP-1细胞。结果：所有动物均存活，处死时静脉通畅。以1.10（9）滴度的AD腔内暴露导致在2天时内皮几乎完全转导，到第14天时没有可检测的表达。在相同滴度的感染性颗粒下，AAV在2至7天的转基因表达明显较少，但在2周时有所改善并持续至40天。AD暴露后2天MA显著增加（2.7倍vs对照，*P < .002）; AAV治疗对MA没有明显影响。结论：AD介导的基因转移至静脉节段导致稳健的瞬时基因表达，2周后消失。相比之下，AAV介导的基因递送效率较低，但导致延迟发作，持续表达超过30天。AD暴露诱导了静脉表面MA的早期增加，这在AAV处理中没有观察到。目前几代AD和AAV载体对于血管基因治疗具有显著的（尽管不同）限制。
Purpose: Gene transfer offers the potential to modify vein graft biology at the time of surgical implantation. Efficiency of gene delivery, stability of expression, and host responses are critical parameters for candidate vectors. We compared the effects of intraluminal exposure with adenovirus (AD) and adeno-associated virus (AAV) vectors on transgene expression and monocyte adhesion (MA) in treated vein segments.Methods: Adult New Zealand white rabbits (N = 51) were anesthetized, and the jugular veins were cannulated bilaterally. Veins were gently distended with either vector (2.10(8) to 1.10(10) infective particles/mL) or vehicle (control) for 30 minutes, after which venous flow was restored. AD and AAV vectors encoding for the marker genes P-galactosidase (LacZ) and green fluorescent protein (GFP) were used. Vessels were explanted 2 to 40 days postinfection for analysis of gene expression (X-gal staining, reverse transcriptase-polymerase chain reaction), MA, and immunohistochemistry. Ex vivo adhesion assays used Cr-51-labeled THP-1 cells. Statistical significance was tested by using analysis of variance with a P value less than .05.Results: All animals survived, and all treated veins were patent at sacrifice. Intraluminal exposure to AD at a titer of 1.10(9) resulted in near complete transduction of the endothelium at 2 days, with no detectable expression by day 14. At an equal titer of infectious particles, transgene expression was markedly less for AAV at 2 to 7 days, but improved at 2 weeks and persisted to 40 days. MA was significantly increased 2 days after AD exposure (2.7-fold vs control, *P < .002); AAV treatment had no discernible effect on MA.Conclusion: AD-mediated gene transfer to vein segments resulted in robust, transient gene expression that disappeared after 2 weeks. In comparison, AAV-mediated gene delivery was less efficient, but resulted in delayed onset, persistent expression beyond 30 days. AD exposure induced an early increase in MA to the vein surface that was not seen with AAV treatment. Current generations of both AD and AAV vectors have significant, albeit different, limitations for vascular gene therapy.