Department of Pediatrics, Division of Pediatric Neurology and Metabolism, Ghent University Hospital, Ghent 9000, Belgium.
Hum Gene Ther. 2013 Feb;24(2):209-19. doi: 10.1089/hum.2012.107.
Giant axonal neuropathy (GAN) is caused by loss of function of the gigaxonin protein. On a cellular level GAN is characterized by intermediate filament (IF) aggregation, leading to a progressive and fatal peripheral neuropathy in humans. This study sought to determine if re-introduction of the GAN gene into GAN-deficient cells and mice would restore proper cytoskeleton IF homeostasis. Treatment of primary skin fibroblast cultures from three different GAN patients with an adeno-associated virus type 2 (AAV2) vector containing a normal human GAN transgene significantly reduced the number of cells displaying vimentin IF aggregates. A proteomic analysis of these treated cells was also performed, wherein the abundance of 32 of 780 identified proteins significantly changed in response to gigaxonin gene transfer. While 29 of these responding proteins have not been directly described in association with gigaxonin, three were previously identified as being disregulated in GAN and were now shifted toward normal levels. To assess the potential application of this approach in vivo and eventually in humans, GAN mice received an intracisternal injection of an AAV9/GAN vector to globally deliver the GAN gene to the brainstem and spinal cord. The treated mice showed a nearly complete clearance of peripherin IF accumulations at 3 weeks post-injection. These studies demonstrate that gigaxonin gene transfer can reverse the cellular IF aggregate pathology associated with GAN.
巨轴索神经病（GAN）是由巨轴索蛋白功能丧失引起的。在细胞水平上，GAN 的特征是中间丝（IF）聚集，导致人类进行性和致命的周围神经病。本研究旨在确定将 GAN 基因重新引入 GAN 缺陷细胞和小鼠中是否会恢复适当的细胞骨架 IF 动态平衡。用含有正常人类 GAN 转基因的腺相关病毒 2 型（AAV2）载体处理来自三名不同 GAN 患者的原代皮肤成纤维细胞培养物，显著减少了显示波形蛋白 IF 聚集的细胞数量。还对这些经处理的细胞进行了蛋白质组学分析，其中 780 种鉴定蛋白中有 32 种的丰度因 gigaxonin 基因转移而显著改变。虽然其中 29 种这些反应蛋白与 gigaxonin 没有直接关联，但有三种以前被鉴定为 GAN 失调的蛋白，现在向正常水平转移。为了评估这种方法在体内（最终在人类中）的潜在应用，GAN 小鼠接受了 AAV9/GAN 载体的脑室内注射，以将 GAN 基因全局递送至脑干和脊髓。治疗后的小鼠在注射后 3 周显示出周围神经纤维蛋白积累的几乎完全清除。这些研究表明，巨轴索蛋白基因转移可以逆转与 GAN 相关的细胞 IF 聚集病理学。
