Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection
Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection
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DOI：
10.1038/mt.2008.76
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发表时间：
2008-06-01
影响因子：
12.4
通讯作者：
Rabinowitz, Joseph E.
中科院分区：
文献类型：
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作者：
Zincarelli, Carmela;Soltys, Stephen;Rabinowitz, Joseph E.
本研究检测了腺相关病毒（AAV）假型1-9在雄性小鼠尾静脉（TV）注射后的转基因表达和生物分布。使用巨细胞病毒（CMV）-荧光素酶转基因，在整个实验中追踪每只动物中表达的时间过程。在TV注射后7、14、29、56和100天对动物进行成像。记录每只动物发出的光子总数，以检查每种假型病毒的表达水平和动力学。生物发光成像显示三个表达水平：（i）低表达组，AAV 2、3、4和5;（ii）中等表达组，AAV 1、6和8;以及（iii）高表达组，AAV 7和9。此外，成像揭示了两类动力学：（i）AAV 1、6、7、8和9的快速发作;和（ii）AAV 2、3、4和5的缓慢发作。我们接下来评估了解剖组织中的蛋白质表达和病毒基因组拷贝数。AAV 9具有最好的病毒基因组分布和最高的蛋白水平。AAV 7蛋白和基因组拷贝数与肝脏中的AAV 9相当。最令人惊讶的是，AAV 4在肺和肾中显示出最大数量的基因组拷贝，并且在心脏中显示出高拷贝数。在心脏、肝脏和骨骼肌中观察到AAV 6表达，基因组分布证实了这些观察结果。
This study examines transgene expression and biodistribution of adeno-associated virus ( AAV) pseudotyped 1-9 after tail vein ( TV) injection in male mice. Using a cytomegalovirus ( CMV)-luciferase transgene, the time-course of expression in each animal was tracked throughout the experiment. The animals were imaged at 7, 14, 29, 56, and 100 days after the TV injection. The total number of photons emitted from each animal was recorded, allowing examination of expression level and kinetics for each pseudotyped virus. The bioluminescence imaging revealed three expression levels ( i) low-expression group, AAV2, 3, 4, and 5; ( ii) moderate-expression group, AAV1, 6, and 8; and ( iii) high-expression group, AAV7 and 9. In addition, imaging revealed two classes of kinetics ( i) rapid-onset, for AAV1, 6, 7, 8, and 9; and ( ii) slow-onset, for AAV2, 3, 4, and 5. We next evaluated protein expression and viral genome copy numbers in dissected tissues. AAV9 had the best viral genome distribution and highest protein levels. The AAV7 protein and genome copy numbers were comparable to those of AAV9 in the liver. Most surprisingly, AAV4 showed the greatest number of genome copies in lung and kidney, and a high copy number in the heart. AAV6 expression was observed in the heart, liver, and skeletal muscle, and the genome distribution corroborated these observations.