Gene Therapy for Retinal Degeneration using Home-grown Gene Transfer Vectors-Aim for Clinical Trial-

1.基因治疗视网膜变性动物模型的疗效评价1)新的基因治疗技术的发展我们研制了携带人成纤维细胞生长因子-2或同时携带人成纤维细胞生长因子和人成纤维细胞生长因子的新型猴免疫缺陷病毒载体(SIV-FGF2或SIV-DUAL)。2)对两种视网膜变性动物模型(RCS大鼠和RDS小鼠)进行了SIV-hPEDF和SIV-hFGF-2的协同治疗。2.利用神经前体细胞建立了新的基因治疗策略。1)建立了从眼睫状体分离神经前体细胞的培养方法。我们发现了一种新的发散机制(重编程系统)，它支持睫状体来源细胞通过球体的神经分化。来自睫状体的巢蛋白阴性上皮样细胞开始表达巢蛋白2)我们证实了通过SIV载体3将基因从眼部睫状体转移到神经前体细胞。我们评估了我们的第三代SIV载体将基因转移到非人类灵长类视网膜的能力。2)我们认为RPE中VEGF-C和VEGF-D的表达可能改变了眼血管生成，如年龄相关性黄斑变性(AMD)作为血管生成刺激因子。

1. The assessment of the efficacy of gene therapy for the animal models of retinal degeneration1) Development of new gene therapy technologyWe developed the new SIV (simian immunodeficiency virus) vectors carrying hFGF-2 or both hPEDF and hFGF-2 (SIV-FGF-2 or SIV-dual). Moreover, we developed the system for large-scale production of SIV-hPEDF with the aim of the clinical trail.2) We showed the synergistic therapeutic effect using SIV-hPEDF and SIV-hFGF-2 for two animal models of retinal degeneration (RCS rats and rds mice)2. The establishment of new gene therapy strategy by use of neural progenitor cells1) Development of the culture method to isolate neural progenitor cells from ciliary body of the eyeWe identified a new and divergent mechanism (a reprogramming system) underlying the neural differentiation of ciliary body-derived cells via sphere formation. The nestin-negative epithelial-like cells from ciliary body came to express nestin.2) We demonstrated the efficient gene transfer to neural progenitor cells from ciliary body of the eye via SIV vector3. Remains1) We evaluated the ability of our 3rd generation SIV vectors to transfer genes into nonhuman primate retinas.2) We suggested the possibility that VEGF-C and VEGF-D expression in RPE modify the ocular angiogenesis, such as age-related macular degeneration (AMD) as angiogenic stimulators.