Novel bioreducible polymer-based delivery platform for intravitreal gene transfer to retina

项目总结 我们在这项提案中的主要目标是开发一种基于聚合物的合成基因传递平台，用于基因治疗 获得性和/或遗传性视网膜和视神经疾病。病毒载体，特别是腺相关病毒 (AAV)，到目前为止一直是首选的基因传递载体，但几个固有的缺点需要 能够适应日益增长的对新基因治疗产品的需求的替代方案。这些措施包括 包装容量有限，病媒特异性免疫反应，以及极高的成本。此外，视网膜下 尽管注射被广泛使用，并提供了直接进入视网膜外层的途径，但它构成了几个 缺点，如光感受器损伤和有限的地形治疗覆盖范围。为此，我们建议 开发和评价新型环境敏感的聚合物基基因递送纳米粒 玻璃体内基因输送到视网膜，用于广泛的视网膜基因治疗，同时避免上述- 提到了限制。具体地说，我们的平台将旨在克服关键的细胞外障碍，包括 玻璃体凝胶和内界膜，以实现广泛的视网膜基因转移，并促进 增强型转染剂的胞内逃逸和治疗性核酸有效载荷的释放 视网膜细胞。重要的是，我们平台的非免疫原性将允许多个给药机构放大 转基因表达水平，潜在地匹配或超过临床可达到的疗效 经过验证的病毒载体。值得注意的是，尽管非病毒基因传递系统不容易表现出特定的细胞 趋向性、细胞特异性治疗性转基因表达可以通过使用细胞特异性启动子来实现，如果 想要。在视网膜中提供广泛和安全的转基因表达的合成基因递送系统可以 具有广泛的临床应用价值。

PROJECT SUMMARY Our primary goal in this proposal is to develop a polymer-based synthetic gene delivery platform for gene therapy of acquired and/or inherited retinal and optic nerve disorders. Viral vectors, particularly adeno-associated virus (AAV), have been the gene delivery vector of choice to date but several inherent shortcomings necessitate an alternative that can accommodate the ever-increasingly need for novel gene therapy products. These include limited packaging capacity, vector-specific immune responses, and extremely high cost. In addition, subretinal injection, despite being widely practiced and affording direct access to the outer retinal layer, poses several drawbacks such as photoreceptor trauma and limited topographic therapeutic coverage. To this end, we propose to develop and evaluate novel environmentally-sensitive polymer-based gene delivery nanoparticles (NPs) for intravitreal gene delivery to the retina for broad retinal gene therapy applications while avoiding the above- mentioned limitations. Specifically, our platform will be designed to overcome key extracellular barriers, including the vitreous gel and inner limiting membrane, to enable widespread retinal gene transfer, and to promote endosomal escape and intracellular release of therapeutic nucleic acid payloads for enhanced transfection of retinal cells. Importantly, non-immunogenic nature of our platform will allow multiple administrations to amplify the level of transgene expression, potentially matching or surpassing the efficacy achievable by clinically validated viral vectors. Of note, although non-viral gene delivery systems do not readily exhibit a specific cell tropism, cell-specific therapeutic transgene expression can be achieved with the use of cell-specific promoters if desired. Synthetic gene delivery systems that provide extensive and safe transgene expression in retina can be of broad clinical utility.