Su H, Huang Y, Takagawa J, Barcena A, Arakawa-Hoyt J, Ye J, Grossman W, Kan Y W
Cardiovascular Research Institute, University of California-San Francisco, 513 Parnassus Avenue, San Francisco, CA 94143, USA.
Gene Ther. 2006 Nov;13(21):1495-502. doi: 10.1038/sj.gt.3302787. Epub 2006 Jun 15.
Adeno-associated viral vectors (AAV) are attractive tool for gene therapy for coronary artery disease. However, gene expression in myocardium mediated by AAV serotype 2 (AAV2) does not peak until 4-6 weeks after gene transfer. This delayed gene expression may reduce its therapeutic potential for acute cardiac infarction. To determine whether earlier gene expression and better therapeutic effect could be achieved using a different serotype, CMV promoter driving the EPO gene (AAV-EPO) was packaged into AAV serotypes 1-5 capsids and injected into mouse myocardium. EPO expression was studied by measuring the hematocrits and EPO mRNA. After we found that AAV1 mediates the highest gene expression after 4 days of gene transduction, AAV-LacZ (CMV promoter driving LacZ gene expression) and MLCVEGF (hypoxia-inducible and cardiac-specific VEGF expression) were packaged into AAV1 and 2 capsids. LacZ expression was detected in AAV1-LacZ but not in AAV2-LacZ-injected hearts 1 day after vector injection. Compared to AAV2-MLCVEGF that mediated no significant VEGF expression, AAV1-MLCVEGF mediated 13.7-fold induction of VEGF expression in ischemic hearts 4 days after gene transduction and resulted in more neovasculatures, better cardiac function and less myocardial fibrosis. Thus, AAV1 mediates earlier and higher transgene expression in myocardium and better therapeutic effects.
腺相关病毒载体（AAV）是用于冠心病基因治疗的有吸引力的工具。然而，由2型腺相关病毒（AAV2）介导的心肌中的基因表达直到基因转移后4至6周才达到峰值。这种延迟的基因表达可能会降低其对急性心肌梗死的治疗潜力。为了确定使用不同血清型是否可以实现更早的基因表达和更好的治疗效果，将驱动EPO基因（AAV-EPO）的巨细胞病毒启动子包装到AAV血清型1-5衣壳中，并注射到小鼠心肌中。通过测量血细胞比容和EPO mRNA来研究EPO表达。在我们发现AAV1在基因转导4天后介导最高的基因表达后，将AAV-LacZ（驱动LacZ基因表达的CMV启动子）和MLCVEGF（缺氧诱导和心脏特异性VEGF表达）包装到AAV1和2衣壳中。在载体注射1天后，在注射AAV1-LacZ的心脏中检测到LacZ表达，但在注射AAV2-LacZ的心脏中未检测到。与未介导显著VEGF表达的AAV2-MLCVEGF相比，AAV1-MLCVEGF在基因转导4天后在缺血心脏中介导了13.7倍的VEGF表达诱导，并导致更多的新生血管、更好的心脏功能和更少的心肌纤维化。因此，AAV1在心肌中介导更早、更高的转基因表达和更好的治疗效果。
