Engineered Lentiviral Vector Targeting Astrocytes In Vivo
Engineered Lentiviral Vector Targeting Astrocytes In Vivo
复制标题
DOI：
10.1002/glia.20795
复制
发表时间：
2009-04-15
期刊：
影响因子：
6.2
通讯作者：
Deglon, Nicole
中科院分区：
文献类型：
--
作者：
Colin, Angelique;Faideau, Mathilde;Deglon, Nicole
星形胶质细胞参与大脑的关键生理过程，如谷氨酸能传递和能量代谢，在神经退行性疾病中经常改变。星形胶质细胞的靶向基因表达是评估这些细胞在生理过程中的作用和开发新的治疗策略所必需的。然而，目前用于中枢神经系统(CNS)基因转移的病毒载体大多具有高度的神经亲和性。我们使用Mokola伪分型来改变慢病毒载体对星形胶质细胞的导向，并使用miRNA去靶向策略来消除神经细胞中的残留表达。在原代培养中，我们证明了整合神经元特异性miR124的目标序列有效地在转录后取消了神经元中的转基因表达。LacZ报告基因在星形胶质细胞中的靶向性表达在成年小鼠的海马体、纹状体和小脑中实现。作为原理的证明，这种新的慢病毒载体被用来在体内过表达或下调(RNA干扰)胶质谷氨酸转运体GLAST到纹状体星形胶质细胞。这些载体为中枢神经系统中特定细胞类型的基因转移提供了新的机会。(C)2008年Wiley-Liss，Inc.
Astrocytes are involved in key physiological brain processes, such as glutamatergic transmission and energy metabolism, often altered in neurodegenerative diseases. Targeted gene expression in astrocytes is needed to assess the contribution of these cells to physiological processes and for the development of new therapeutic strategies. However, most of the viral vectors currently used for gene transfer in the central nervous system (CNS) are highly neurotropic. We used mokola pseudotyping to shift the tropism of lentiviral vectors toward astrocytes and a detargeting strategy with miRNA to eliminate residual expression in neuronal cells. In primary cultures, we showed that incorporating target sequences for the neuron-specific miR124 effectively abolished transgene expression in neurons post-transcriptionally. Targeted expression of the LacZ reporter gene in astrocytes was achieved in the hippocampus, striatum, and cerebellum of the adult mouse in vivo. As a proof-of-principle, this new lentiviral vector was used to either overexpress or downregulate (RNA interference) the glial glutamate transporter GLAST into striatal astrocytes in vivo. These vectors provide new opportunities for cell type-specific gene transfer in the CNS. (C) 2008 Wiley-Liss, Inc.