Roddy Gavin W, Yasumura Douglas, Matthes Michael T, Alavi Marcel V, Boye Sanford L, Rosa Robert H, Fautsch Michael P, Hauswirth William W, LaVail Matthew M
Department of Ophthalmology, Mayo Clinic, Rochester, MN 55905, USA.
Department of Ophthalmology, University of California, San Francisco, CA 94143, USA.
Exp Eye Res. 2017 Dec;165:175-181. doi: 10.1016/j.exer.2017.09.011. Epub 2017 Sep 30.
Retinal degenerations, including age-related macular degeneration and the retinitis pigmentosa family of diseases, are among the leading causes of legal blindness in the United States. We previously found that Stanniocalcin-1 (STC-1) reduced photoreceptor loss in the S334ter-3 and Royal College of Surgeons rat models of retinal degeneration. The results were attributed in part to a reduction in oxidative stress. Herein, we tested the hypothesis that long-term delivery of STC-1 would provide therapeutic rescue in more chronic models of retinal degeneration. To achieve sustained delivery, we produced an adeno-associated virus (AAV) construct to express STC-1 (AAV-STC-1) under the control of a retinal ganglion cell targeting promoter human synapsin 1 (hSYN1). AAV-STC-1 was injected intravitreally into the P23H-1 and S334ter-4 rhodopsin transgenic rats at postnatal day 10. Tissues were collected at postnatal day 120 for confirmation of STC-1 overexpression and histologic and molecular analysis. Electroretinography (ERG) was performed in a cohort of animals at that time. Overexpression of STC-1 resulted in a significant preservation of photoreceptors as assessed by outer nuclear thickness in the P23H-1 (P < 0.05) and the S334ter-4 (P < 0.005) models compared to controls. Additionally, retinal function was significantly improved in the P23H-1 model with overexpressed STC-1 as assessed by ERG analysis (scotopic b-wave P < 0.005 and photopic b-wave P < 0.05). Microarray analysis identified common downstream gene expression changes that occurred in both models. Genes of interest based on their function were selected for validation by quantitative real-time PCR and were significantly increased in the S334ter-4 model.
视网膜变性，包括年龄相关性黄斑变性和色素性视网膜炎家族性疾病，是美国法定失明的主要原因之一。我们之前发现，在视网膜变性的S334ter-3和皇家外科医学院大鼠模型中，鲽钙蛋白-1（STC-1）可减少光感受器的损失。部分结果归因于氧化应激的降低。在此，我们测试了以下假设：长期递送STC-1将在更慢性的视网膜变性模型中提供治疗性挽救。为了实现持续递送，我们构建了一种腺相关病毒（AAV）载体，使其在视网膜神经节细胞靶向启动子人突触素1（hSYN1）的控制下表达STC-1（AAV-STC-1）。在出生后第10天，将AAV-STC-1玻璃体内注射到P23H-1和S334ter-4视紫红质转基因大鼠体内。在出生后第120天收集组织，以确认STC-1的过表达并进行组织学和分子分析。此时对一组动物进行视网膜电图（ERG）检查。与对照组相比，通过P23H-1（P < 0.05）和S334ter-4（P < 0.005）模型中外核层厚度评估，STC-1的过表达导致光感受器显著保留。此外，通过ERG分析评估，在过表达STC-1的P23H-1模型中，视网膜功能显著改善（暗视b波P < 0.005，明视b波P < 0.05）。微阵列分析确定了两个模型中共同发生的下游基因表达变化。根据其功能选择感兴趣的基因通过定量实时PCR进行验证，在S334ter-4模型中这些基因显著增加。
