Lentiviral-Mediated Silencing of Glial Fibrillary Acidic Protein and Vimentin Promotes Anatomical Plasticity and Functional Recovery After Spinal Cord Injury
Lentiviral-Mediated Silencing of Glial Fibrillary Acidic Protein and Vimentin Promotes Anatomical Plasticity and Functional Recovery After Spinal Cord Injury
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DOI：
10.1002/jnr.23468
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发表时间：
2015-01-01
影响因子：
4.2
通讯作者：
Privat, Alain
中科院分区：
文献类型：
--
作者：
Desclaux, Mathieu;Perrin, Florence E.;Privat, Alain
在脊髓损伤（SCI）中，除了其他因素之外，功能恢复的缺乏和自发轴突再生的缺乏归因于形成物理和化学屏障的胶质瘢痕的形成。胶质瘢痕主要由过表达两种中间丝蛋白--胶质细胞酸性蛋白（GFAP）和波形蛋白（Vim）的反应性星形胶质细胞组成。为了促进脊髓损伤后备用轴突的再生和发芽，并将其转化为临床目标，我们设计了一种基于RNA干扰（RNAi）介导的GFAP和Vim抑制的体内基因转移策略，以减少SCI后胶质瘢痕形成。我们首先表明，在SCI小鼠模型中直接注射表达针对GFAP和Vim的短发夹RNA（shRNA）的慢病毒载体可以有效和特异性地靶向星形胶质细胞。然后，我们证明慢病毒介导的shGFAP和shVIM的稳定表达导致星形胶质细胞增生的强烈减少，改善功能性运动恢复，并促进轴突再生和备用轴突发芽。因此，这项研究举例说明了非神经元环境如何成为受损中枢神经系统内促进轴突再生（和发芽）的主要靶点，并验证了慢病毒介导的RNAi在SCI中的应用。(c)2014 Wiley Periodicals，Inc.
In spinal cord injury (SCI), absence of functional recovery and lack of spontaneous axonal regeneration are attributed, among other factors, to the formation of a glial scar that forms both physical and chemical barriers. The glial scar is composed mainly of reactive astrocytes that overexpress two intermediate filament proteins, glial fibrillary acidic protein (GFAP) and vimentin (VIM). To promote regeneration and sprouting of spared axons after spinal cord trauma and with the objective of translation to clinics, we designed an original in vivo gene transfer strategy to reduce glial scar formation after SCI, based on the RNA interference (RNAi)-mediated inhibition of GFAP and VIM. We first show that direct injection of lentiviral vectors expressing short hairpin RNA (shRNA) against GFAP and VIM in a mouse model of SCI allows efficient and specific targeting of astrocytes. We then demonstrate that the lentiviral-mediated and stable expression of shGFAP and shVIM leads to a strong reduction of astrogliosis, improves functional motor recovery, and promotes axonal regrowth and sprouting of spared axons. This study thus examplifies how the nonneuronal environment might be a major target within the lesioned central nervous system to promote axonal regeneration (and sprouting) and validates the use of lentiviral-mediated RNAi in SCI. (c) 2014 Wiley Periodicals, Inc.