McGee Sanftner L H, Abel H, Hauswirth W W, Flannery J G
Department of Vision Science, University of California, Berkeley, California 94720, USA.
Mol Ther. 2001 Dec;4(6):622-9. doi: 10.1006/mthe.2001.0498.
We designed experiments to evaluate the therapeutic potential of glial cell line derived neurotrophic factor (GDNF) to rescue photoreceptors from genetically determined cell death. Gene transfer of the neurotrophic factor to the retina was achieved via a recombinant adeno-associated virus (rAAV) vector containing the chicken beta-actin promoter/immediate early cytomegalovirus enhancer (CBA) driving the human GDNF gene. We delivered AAV-CBA-GDNF to the retinas of an animal model of retinitis pigmentosa, the TgN S334ter-4 rhodopsin line of transgenic rats. Immunohistochemical studies localized AAV-CBA-GDNF-derived recombinant protein to cell bodies, inner segments, and outer segments of photoreceptor cells as well as to retinal pigment epithelial cells. We assessed the effect of viral delivery by morphometric and electroretinographic analysis. These experiments showed that GDNF vector treatment leads to increased rod photoreceptor survival as indicated by morphometric analysis of outer nuclear layer thickness. AAV-CBA-GDNF-treated retinas also demonstrated functional improvement by the substantially increased amplitude of electroretinograms. AAV-CBA-GDNF delivery had a significant rescue effect on photoreceptor degeneration in this animal model.
我们设计了实验来评估胶质细胞源性神经营养因子（GDNF）从基因决定的细胞死亡中拯救光感受器的治疗潜力。通过含有驱动人类GDNF基因的鸡β-肌动蛋白启动子/立即早期巨细胞病毒增强子（CBA）的重组腺相关病毒（rAAV）载体，将神经营养因子基因转移至视网膜。我们将AAV-CBA-GDNF注射到视网膜色素变性动物模型——TgN S334ter-4视紫红质转基因大鼠的视网膜中。免疫组织化学研究将AAV-CBA-GDNF衍生的重组蛋白定位到光感受器细胞的细胞体、内节和外节以及视网膜色素上皮细胞。我们通过形态测量和视网膜电图分析评估病毒注射的效果。这些实验表明，如通过外核层厚度的形态测量分析所示，GDNF载体治疗可导致视杆光感受器存活率增加。经AAV-CBA-GDNF治疗的视网膜通过视网膜电图幅度的大幅增加也显示出功能改善。在该动物模型中，AAV-CBA-GDNF注射对光感受器变性具有显著的拯救作用。
