Transduction of the Central Nervous System After Intracerebroventricular Injection of Adeno-Associated Viral Vectors in Neonatal and Juvenile Mice
Transduction of the Central Nervous System After Intracerebroventricular Injection of Adeno-Associated Viral Vectors in Neonatal and Juvenile Mice
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DOI：
10.1089/hgtb.2013.076
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发表时间：
2013-08-01
影响因子：
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通讯作者：
Hampson, David R.
中科院分区：
文献类型：
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作者：
Gholizadeh, Shervin;Tharmalingam, Sujeenthar;Hampson, David R.
几种影响中枢神经系统的神经发育和神经退行性疾病有可能通过病毒载体介导的基因转移治疗。腺相关病毒（AAV）载体具有高度的安全性、有效性和稳定性，已被用于临床试验。影响AAV介导的转基因趋向性、表达水平和细胞类型特异性的主要因素包括不同AAV血清型的衣壳化、启动子的选择和载体给药的时机。在这项研究中，我们评估了用血清型9病毒衣壳伪型的单链AAV2载体（AAV2/9）在小鼠脑室内注射后转导大脑和靶基因表达到特定细胞类型的能力。在巨细胞病毒（CMV）启动子或神经元特异性突触素-1启动子的驱动下，编码增强绿色荧光蛋白（eGFP）报告基因的AAV2/9载体滴度匹配，于出生后第5天（新生）或21天（幼年）注射到C57/BL6小鼠的侧脑室。注射后25天，用免疫细胞化学和共聚焦显微镜分析脑切片。在新生儿和青少年注射病毒载体后，eGFP免疫组化显示转导贯穿整个大脑，包括纹状体、海马、大脑皮层和小脑，但具有不同的细胞特异性基因表达模式。在出生后第5天，携带CMV启动子的载体在星形胶质细胞中表达eGFP，扩大了aav在模拟和治疗涉及胶质细胞病理的疾病方面的有用性。相反，在出生后第21天注射AAV2/9-CMV-eGFP导致神经元优先转导。AAV2/9-eGFP与突触素-1启动子一起在出生后第5天或21天使用可导致广泛的神经元转导。这些结果概述了通过出生后早期直接给予AAV载体将基因传递到神经系统的有效方法和工具。我们的研究结果强调了启动子选择和给药年龄对大脑中AAV转导的强度、分布和细胞类型特异性的重要性。
Several neurodevelopmental and neurodegenerative disorders affecting the central nervous system are potentially treatable via viral vector-mediated gene transfer. Adeno-associated viral (AAV) vectors have been used in clinical trials because of their desirable properties including a high degree of safety, efficacy, and stability. Major factors affecting tropism, expression level, and cell type specificity of AAV-mediated transgenes include en-capsidation of different AAV serotypes, promoter selection, and the timing of vector administration. In this study, we evaluated the ability of single-stranded AAV2 vectors pseudotyped with viral capsids from serotype 9 (AAV2/9) to transduce the brain and target gene expression to specific cell types after intracerebroventricular injection into mice. Titer-matched AAV2/9 vectors encoding the enhanced green fluorescent protein (eGFP) reporter, driven by the cytomegalovirus (CMV) promoter, or the neuron-specific synapsin-1 promoter, were injected bilaterally into the lateral ventricles of C57/BL6 mice on postnatal day 5 (neonatal) or 21 (juvenile). Brain sections were analyzed 25 days after injection, using immunocytochemistry and confocal microscopy. eGFP immunohistochemistry after neonatal and juvenile administration of viral vectors revealed transduction throughout the brain including the striatum, hippocampus, cerebral cortex, and cerebellum, but with different patterns of cell-specific gene expression. eGFP expression was seen in astrocytes after treatment on postnatal day 5 with vectors carrying the CMV promoter, expanding the usefulness of AAVs for modeling and treating diseases involving glial cell pathology. In contrast, injection of AAV2/9-CMV-eGFP on postnatal day 21 resulted in preferential transduction of neurons. Administration of AAV2/9-eGFP with the synapsin-1 promoter on either postnatal day 5 or 21 resulted in widespread neuronal transduction. These results outline efficient methods and tools for gene delivery to the nervous system by direct, early postnatal administration of AAV vectors. Our findings highlight the importance of promoter selection and age of administration on the intensity, distribution, and cell type specificity of AAV transduction in the brain.