Comparative Transduction Efficiency of AAV Vector Serotypes 1-6 in the Substantia Nigra and Striatum of the Primate Brain
Comparative Transduction Efficiency of AAV Vector Serotypes 1-6 in the Substantia Nigra and Striatum of the Primate Brain
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DOI：
10.1038/mt.2009.286
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发表时间：
2010-03-01
影响因子：
12.4
通讯作者：
Redmond, D. Eugene, Jr.
中科院分区：
文献类型：
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作者：
Markakis, Eleni A.;Vives, Kenneth P.;Redmond, D. Eugene, Jr.
来源于腺相关病毒（AAV）的载体是用于体内神经细胞转导的有希望的候选者，因为它们是非致病性的并且在中枢神经系统中实现长期转导。AAV血清型2（AAV 2）是临床试验中最广泛使用的AAV载体，这主要基于其在啮齿动物和灵长类动物脑中激活神经细胞的能力。先前在啮齿动物中的工作表明，其他血清型可能更有效;然而，尚未在灵长类动物脑中进行载体转导效率的系统评价。在这项研究中，以相当的滴度产生具有增强的绿色荧光蛋白（GFP）报告基因的血清型1 - 6的AAV病毒载体，并将其等量注射到绿猴脑中。将载体注射液置于黑质（SN）和尾状核（CD）中。注射后一个月，进行GFP免疫组化，并使用无偏体视学计算GFP（+）细胞总数。AAV5是最有效的载体，不仅比任何其他血清型转导显著更多的细胞，而且转导NeuN（+）和胶质细胞酸性蛋白阳性（GFAP（+））细胞。这些结果表明，在向灵长类动物脑的黑质纹状体系统递送潜在治疗性转基因方面，AAV5是比AAV2更有效的载体。
Vectors derived from adeno-associated virus (AAV) are promising candidates for neural cell transduction in vivo because they are nonpathogenic and achieve long-term transduction in the central nervous system. AAV serotype 2 (AAV2) is the most widely used AAV vector in clinical trials based largely on its ability to transduce neural cells in the rodent and primate brain. Prior work in rodents suggests that other serotypes might be more efficient; however, a systematic evaluation of vector transduction efficiency has not yet been performed in the primate brain. In this study, AAV viral vectors of serotypes 1-6 with an enhanced green-fluorescent protein (GFP) reporter gene were generated at comparable titers, and injected in equal amounts into the brains of Chlorocebus sabaeus. Vector injections were placed in the substantia nigra (SN) and the caudate nucleus (CD). One month after injection, immunohistochemistry for GFP was performed and the total number of GFP(+) cells was calculated using unbiased stereology. AAV5 was the most efficient vector, not only transducing significantly more cells than any other serotype, but also transducing both NeuN(+) and glial-fibrillary-acidic protein positive (GFAP(+)) cells. These results suggest that AAV5 is a more effective vector than AAV2 at delivering potentially therapeutic transgenes to the nigrostriatal system of the primate brain.