Lentiviral vector-mediated gene transfer in adult mouse photoreceptors is impaired by the presence of a physical barrier
Lentiviral vector-mediated gene transfer in adult mouse photoreceptors is impaired by the presence of a physical barrier
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DOI：
10.1038/sj.gt.3302485
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发表时间：
2005-06-01
期刊：
影响因子：
5.1
通讯作者：
Arsenijevic, Y
中科院分区：
文献类型：
--
作者：
Grüter, O;Kostic, C;Arsenijevic, Y
基因转移为退行性眼病的治疗提供了巨大的希望。慢病毒载体能够在生命的最初几天有效地刺激小鼠光感受器，但它们在成年期对光感受器的作用很差。在这里，我们研究了物理障碍是否是造成这种损害的原因。以前的研究已经描述了酶的能力，如软骨素酶ABC和神经氨酸酶X，以修改结构的interphotoreceptor矩阵（IPM）时，视网膜下注射。考虑到IPM作为可能降低感光细胞转导的物理屏障，我们将不同的酶与慢病毒载体制剂同时注射到成年小鼠的视网膜下腔中，以通过使IPM脆弱化来增加病毒转导。视网膜下注射神经氨酸酶X和软骨素酶ABC通过分别揭示或减少光感受器上的花生凝集素位点来诱导IPM中的修饰。同时视网膜下注射神经氨酸酶X与驱动光感受器中报告基因表达的慢病毒载体，显著增加了转导细胞的数量（约5倍）。酶处理后，载体在色素上皮和光感受器之间的扩散似乎有利于慢病毒载体转导。这种靶向IPM的方法可能有助于设计新的策略来改善基因递送到成人光感受器中。
Gene transfer offers a substantial promise for the therapy of degenerative ocular diseases. Lentiviral vectors have the ability to efficiently transduce murine photoreceptors during the first days of life, but they are poorly effective on photoreceptors during adulthood. Here, we studied whether a physical barrier was responsible for this impairment. Previous studies have described the capacity of enzymes, such as chondroitinase ABC and neuraminidase X, to modify the structure of the interphotoreceptor matrix (IPM) when subretinally injected. Considering the IPM as a physical barrier that may decrease photoreceptor transduction, we injected different enzymes into the subretinal space of the adult mouse simultaneously with the lentiviral vector preparation, to increase viral transduction by fragilizing the IPM. Subretinal injection of neuraminidase X and chondroitinase ABC induces modifications in the IPM by, respectively, revealing or decreasing peanut agglutinin sites on photoreceptors. The simultaneous subretinal injection of neuraminidase X with a lentiviral vector driving the expression of a reporter gene in the photoreceptors increases the number of transduced cells significantly ( around five-fold). After the enzyme treatment, the diffusion of the vector between the pigmented epithelium and the photoreceptors appears to facilitate the lentiviral vector transduction. Such approach targeting the IPM may help to design new strategies to improve gene delivery into the adult photoreceptors.