Long-term transgene expression from plasmid DNA gene therapy vectors is negatively affected by CpG dinucleotides
Long-term transgene expression from plasmid DNA gene therapy vectors is negatively affected by CpG dinucleotides
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DOI：
10.1016/j.ymthe.2004.04.018
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发表时间：
2004-08-01
影响因子：
12.4
通讯作者：
Scheule, RK
中科院分区：
文献类型：
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作者：
Hodges, BL;Taylor, KM;Scheule, RK
将CpG还原的、基于CMV的编码人(X-半乳糖苷酶A和凝血因子IX)的质粒DNA注射到C57B1/6、BALB/c和CD1小鼠体内，采用基于流体力学的递送方法(PDNA)，并对基因表达进行了6个月的监测。线性化和超螺旋pDNA被比较了它们支持长期表达和对转基因产物产生免疫反应的能力。在所有小鼠品系中，编码α-半乳糖苷酶A和因子IX的超螺旋CpG还原的PDNA比它们的超螺旋CpG完全类似物产生更高和更持久的循环基因产物水平。线性化超螺旋CpG还原的PDNA不会显著增加循环基因产物的水平，超过超螺旋CpG还原的PDNA所能达到的水平。与其超螺旋CpG全长类似物相比，线性化的超螺旋CpG全长PDNA载体显著增加了表达，但这种增加是短暂的或不具疗效的。不管载体是什么，肝库的表达不会引起对人α-半乳糖苷酶A或因子IX的显著抗体反应。综上所述，这些数据表明，临床上可接受的以肝脏为靶点的流体力学方法与CpG降低的PDNA载体相结合，可能是血友病、溶酶体储存病或其他需要长期储存来自肝脏的治疗性蛋白表达的疾病患者的可行选择。
CpG-reduced, CMV-based plasmid DNA constructs encoding human (X-galactosidase A and factor IX were injected into C57B1/6, BALB/c, and CD1 mice using hydrodynamics-based delivery of plasmid DNA (pDNA), and gene expression was monitored for 6 months. Linearized and supercoiled pDNAs were compared for their abilities to support long-term expression and to generate immune responses to the transgene product. In all mouse strains supercoiled CpG-reduced pDNA encoding alpha-galactosidase A and factor IX generated higher and more sustained levels of circulating gene product than their supercoiled CpG-replete analogs. Linearizing supercoiled CpG-reduced pDNA did not significantly increase levels of circulating gene product beyond levels supercoiled CpG-reduced pDNA could achieve. Linearizing supercoiled CpG-replete pDNA vectors significantly increased expression compared to their supercoiled CpG-replete analogs, but the increase was short-lived or subtherapeutic. Regardless of vector, liver depot expression did not elicit significant antibody responses to human alpha-galactosidase A or factor IX. Taken together, these data suggest that a clinically acceptable hydrodynamics-based approach targeting the liver combined with CpG-reduced pDNA vectors may represent a viable option for individuals with hemophilia, a lysosomal storage disease, or other disease in which prolonged depot expression of a therapeutic protein from the liver is desirable.