Evaluation and Optimization of the Administration of Recombinant Adeno-Associated Viral Vectors (Serotypes 2/1, 2/2, 2/rh8, 2/9, and 2/rh10) by Convection-Enhanced Delivery to the Striatum
Evaluation and Optimization of the Administration of Recombinant Adeno-Associated Viral Vectors (Serotypes 2/1, 2/2, 2/rh8, 2/9, and 2/rh10) by Convection-Enhanced Delivery to the Striatum
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DOI：
10.1089/hum.2010.129
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发表时间：
2011-02-01
影响因子：
4.2
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中科院分区：
文献类型：
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重组腺相关病毒（rAAV）载体的对流增强递送（CED）是用于将治疗性转基因递送至脑的有前景的方法。在这项研究中，我们系统地研究了载体给药在体内。使用CED将表达增强的绿色荧光蛋白报告基因的rAAV血清型2/1、2/2、2/rh 8、2/9和2/rh 10输注到大鼠和猪的纹状体中。使用体视学方法确定载体分布，如通过分布体积和每次输注后转导细胞的数量所定义的。免疫组织化学用于确定血清型的转导向性，并评估是否存在免疫细胞浸润到脑中。载体分布在血清型之间高度可变。输注速率对矢量分布或组织损伤的发生没有显著影响。对于血清型2/1、2/2和2/rh 10，当载体浓度增加超过10（12）vg/ml时，未观察到载体分布增加。相比之下，对于血清型2/rh 8和2/9，观察到逆行轴突运输高于此阈值浓度。细胞转导主要是神经元的所有血清型，并与低水平的免疫反应。在规划临床试验时，考虑这些观察结果以实现最佳载体给药至关重要。
Convection-enhanced delivery (CED) of recombinant adeno-associated virus (rAAV) vectors is a promising approach for delivery of therapeutic transgenes to the brain. In this study we have systematically examined vector dosing in vivo. Infusions of rAAV serotypes 2/1, 2/2, 2/rh8, 2/9, and 2/rh10 expressing an enhanced green fluorescent protein reporter gene were undertaken into the striatum of rats and pigs using CED. Vector distribution, as defined by the volume of distribution and number of transduced cells following each infusion, was determined using stereological methods. Immunohistochemistry was used to determine the transductional tropism of serotypes and to evaluate for the presence of immune cell infiltration into the brain. Vector distribution was highly variable between serotypes. Infusion rate had no significant effect on vector distribution or the occurrence of tissue damage. For serotypes 2/1, 2/2 and 2/rh10, as the vector concentration was increased beyond 10(12) vg/ml, no increase in vector distribution was observed. In contrast, for serotypes 2/rh8 and 2/9, retrograde axonal transport was observed above this threshold concentration. Cell transduction was principally neuronal for all serotypes and was associated with a low-level immune response. In planning clinical trials it is critical that these observations are considered in order to achieve optimal vector dosing.