He Xiaoyu, Fu Yidian, Xu Yangfan, Ma Liang, Chai Peiwei, Shi Hanhan, Yao Yizheng, Ge Shengfang, Jia Renbing, Wen Xuyang, Yang Zhi, Fan Xianqun
Department of Ophthalmology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, Shanghai, China.
Invest Ophthalmol Vis Sci. 2025 Jan 2;66(1):6. doi: 10.1167/iovs.66.1.6.
This study aimed to identify a novel recombinant adeno-associated virus (rAAV) capsid variant that can widely transfect the deep retina through intravitreal injection and to assess their effectiveness and safety in gene delivery.
By adopting the sequences of various cell-penetrating peptides and inserting them into the capsid modification region of AAV2, we generated several novel variants. The green fluorescent protein (GFP)-carrying variants were screened following intravitreal injection. Gene therapy experiments were conducted via intravitreal injection of rd1 mice. We validated the therapeutic effects utilizing the pupillary light reflex and visual cliff test. Assessment of retinal structure and Pde6b gene levels in rd1 mice after gene therapy further was confirmed through transcriptome sequencing to validate the gene therapy efficacy.
We observed enhanced transduction and penetration efficiency of the AAV variant AAV2.CPP.21 after intravitreal injection which can target all layers of the retinas. Normal doses of AAV2.CPP.21 administered via intravitreal injection achieved effective gene therapy for retinitis pigmentosa in rd1 mice, with no increased risk of transgenic leakage in peripheral organs.
Our study identified another new, safe, and efficient AAV vector for gene therapy via intravitreal injection for retinal diseases. This new vector holds promise for clinical application and improvement of the efficacy of gene therapy for inherited retinal diseases.
本研究旨在鉴定一种新型重组腺相关病毒（rAAV）衣壳变体，该变体可通过玻璃体内注射广泛转染深层视网膜，并评估其在基因递送中的有效性和安全性。
通过采用各种细胞穿透肽的序列并将其插入AAV2的衣壳修饰区域，我们产生了几种新型变体。玻璃体内注射后筛选携带绿色荧光蛋白（GFP）的变体。通过玻璃体内注射rd1小鼠进行基因治疗实验。我们利用瞳孔光反射和视崖试验验证治疗效果。通过转录组测序进一步证实基因治疗后rd1小鼠的视网膜结构和Pde6b基因水平的评估，以验证基因治疗效果。
我们观察到玻璃体内注射后AAV变体AAV2.CPP.21的转导和穿透效率增强，其可靶向视网膜的所有层。通过玻璃体内注射给予正常剂量的AAV2.CPP.21对rd1小鼠的色素性视网膜炎实现了有效的基因治疗，外周器官中转基因泄漏风险没有增加。
我们的研究鉴定了另一种用于视网膜疾病玻璃体内注射基因治疗的新型、安全且高效的AAV载体。这种新载体有望用于临床应用并提高遗传性视网膜疾病基因治疗的疗效。
