Transendocardial Delivery of AAV6 Results in Highly Efficient and Global Cardiac Gene Transfer in Rhesus Macaques
Transendocardial Delivery of AAV6 Results in Highly Efficient and Global Cardiac Gene Transfer in Rhesus Macaques
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DOI：
10.1089/hum.2011.042
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发表时间：
2011-08-01
影响因子：
4.2
通讯作者：
Sweeney, H. Lee
中科院分区：
文献类型：
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作者：
Gao, Guangping;Bish, Lawrence T.;Sweeney, H. Lee
心脏病是发病率和死亡率的主要原因，心脏基因转移作为一种新的治疗方法具有潜力。我们以前证明了安全和有效的基因转移到狗的心脏，使用经皮transendothelin注射程序，以提供自我互补（sc）腺相关病毒6（AAV 6）载体。在本研究中，我们继续我们的垂直翻译研究，以评估心脏基因转移在非人灵长类动物（NHP）。我们筛选了大约30只成年雄性恒河猴中是否存在针对AAV 6、AAV 8和AAV 9的中和抗体，然后选择了7只针对这三种血清型的抗体滴度低于1/5的猴子。然后将动物随机接受表达增强型绿色荧光蛋白（EGFP）报告基因的scAAV 6（n = 3）、scAAV 8（n = 1）或scAAV 9（n = 3）载体，剂量为5.4 × 10（12）个基因组拷贝/kg，根据我们先前开发的经内皮素注射程序的修改版本进行给药。用scAAV 6治疗的一只动物继发于食管插管而死亡。在基因转移后7天将剩余的动物安乐死，此时收集组织用于分析载体基因组的EGFP表达、组织病理学和生物分布。我们发现，（i）AAV的经内皮细胞递送在NHP中是安全的，（ii）AAV 6和AAV 8以相似的水平提供有效的心脏基因转移，并且上级AAV 9，以及（iii）AAV 6比AAV 8和AAV 9更具心脏特异性。这项NHP研究的结果可能有助于指导开发用于治疗人类心血管疾病的AAV载体。
Heart disease is the leading cause of morbidity and mortality, and cardiac gene transfer has potential as a novel therapeutic approach. We previously demonstrated safe and efficient gene transfer to the canine heart using a percutaneous transendocardial injection procedure to deliver self-complementary (sc) adeno-associated virus 6 (AAV6) vector. In the present study, we proceed with our vertical translation study to evaluate cardiac gene transfer in nonhuman primates (NHPs). We screened approximately 30 adult male rhesus macaques for the presence of neutralizing antibodies against AAV6, AAV8, and AAV9, and then selected seven monkeys whose antibody titers against these three serotypes were lower than 1/5. The animals were then randomized to receive either scAAV6 (n = 3), scAAV8 (n = 1), or scAAV9 (n = 3) vector expressing the enhanced green fluorescent protein (EGFP) reporter gene at a dose of 5.4 x 10(12) genome copies/kg, which was administered according to a modified version of our previously developed transendocardial injection procedure. One animal treated with scAAV6 died secondary to esophageal intubation. The remaining animals were euthanized 7 days after gene transfer, at which time tissue was collected for analysis of EGFP expression, histopathology, and biodistribution of the vector genome. We found that (i) transendocardial delivery of AAV is safe in the NHP, (ii) AAV6 and AAV8 provide efficient cardiac gene transfer at similar levels and are superior to AAV9, and (iii) AAV6 is more cardiac-specific than AAV8 and AAV9. The results of this NHP study may help guide the development AAV vectors for the treatment of cardiovascular disease in humans.