Cellular selectivity of AAV serotypes for gene delivery in neurons and astrocytes by neonatal intracerebroventricular injection.
Cellular selectivity of AAV serotypes for gene delivery in neurons and astrocytes by neonatal intracerebroventricular injection.
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DOI：
10.1371/journal.pone.0188830
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Tjalkens RB
中科院分区：
文献类型：
--
作者：
Hammond SL;Leek AN;Richman EH;Tjalkens RB
非致病性细小病毒，腺相关病毒（AAV），是一种有效的载体，转基因表达在体内，并显示出在临床试验中治疗脑疾病的希望。目前，已经鉴定出超过100种AAV血清型，其衣壳蛋白与特定细胞表面受体的结合能力不同，所述特定细胞表面受体可以覆盖CNS中的不同细胞类型和脑区域。在本研究中，在原代鼠星形胶质细胞和神经元细胞培养物中筛选表达GFP报告基因的多种AAV血清型（AAV 1、AAV 2/1、AAV DJ、AAV 8、AAV DJ 8、AAV 9、AAV DJ 9）的感染性。选择AAV 2/1、AAVDJ 8和AAV 9用于进一步研究它们在不同脑区域和细胞类型中的向性。通过脑室内注射（ICV）向PO-新生小鼠施用每种AAV。然后在感染后3或6周系统地分析脑中各个区域的GFP表达，包括嗅球、纹状体、皮质、海马、黑质（SN）和小脑。细胞计数数据显示，AAV 2/1感染在皮质层中更普遍，但渗透到中脑的程度低于AAVDJ 8和AAV 9。此外，在感染后3周和6周，在体内检查的三种血清型之间的病毒转基因表达的持续性存在差异。因为AAV介导的转基因表达在神经退行性疾病如帕金森病中是感兴趣的，所以我们用显微镜技术（如Vidity组织转化）检查了SN，以鉴定导致星形胶质细胞或多巴胺能神经元中最佳转基因表达的AAV血清型。AAVDJ 8在星形胶质细胞中显示出比AAV 9在SN区域中更大的向性。我们的结论是，ICV注射的结果，在持久的表达病毒编码的转基因时，使用AAV载体和特定的AAV血清型需要选择性地提供转基因的兴趣在星形胶质细胞和神经元的不同脑区域。
The non-pathogenic parvovirus, adeno-associated virus (AAV), is an efficient vector for transgene expression in vivo and shows promise for treatment of brain disorders in clinical trials. Currently, there are more than 100 AAV serotypes identified that differ in the binding capacity of capsid proteins to specific cell surface receptors that can transduce different cell types and brain regions in the CNS. In the current study, multiple AAV serotypes expressing a GFP reporter (AAV1, AAV2/1, AAVDJ, AAV8, AAVDJ8, AAV9, AAVDJ9) were screened for their infectivity in both primary murine astrocyte and neuronal cell cultures. AAV2/1, AAVDJ8 and AAV9 were selected for further investigation of their tropism throughout different brain regions and cell types. Each AAV was administered to P0-neonatal mice via intracerebroventricular injections (ICV). Brains were then systematically analyzed for GFP expression at 3 or 6 weeks post-infection in various regions, including the olfactory bulb, striatum, cortex, hippocampus, substantia nigra (SN) and cerebellum. Cell counting data revealed that AAV2/1 infections were more prevalent in the cortical layers but penetrated to the midbrain less than AAVDJ8 and AAV9. Additionally, there were differences in the persistence of viral transgene expression amongst the three serotypes examined in vivo at 3 and 6 weeks post-infection. Because AAV-mediated transgene expression is of interest in neurodegenerative diseases such as Parkinson’s Disease, we examined the SN with microscopy techniques, such as CLARITY tissue transmutation, to identify AAV serotypes that resulted in optimal transgene expression in either astrocytes or dopaminergic neurons. AAVDJ8 displayed more tropism in astrocytes compared to AAV9 in the SN region. We conclude that ICV injection results in lasting expression of virally encoded transgene when using AAV vectors and that specific AAV serotypes are required to selectively deliver transgenes of interest to different brain regions in both astrocytes and neurons.
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影响因子：
5.4
作者：
Akache, Bassel;Grimm, Dirk;Kay, Mark A.
通讯作者：
Kay, Mark A.
AAV血清型2/1介导的抗炎白细胞介素-10的基因递送增强了APP+PS1小鼠的神经发生和认知功能。
影响因子：
5.1
作者：
通讯作者：
--
DOI：
10.1038/mt.2009.44
发表时间：
2009-05
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
通讯作者：
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影响因子：
48
作者：
Marks, William J., Jr.;Bartus, Raymond T.;Olanow, C. Warren
通讯作者：
Olanow, C. Warren
影响因子：
3.7
作者：
Howard, Douglas B.;Powers, Kathleen;Harvey, Brandon K.
通讯作者：
Harvey, Brandon K.