Department of Ophthalmology, Stanford University School of Medicine, Palo Alto, CA, 94304, USA.
Department of Ophthalmology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, 200120, China.
Adv Sci (Weinh). 2024 Nov;11(42):e2405628. doi: 10.1002/advs.202405628. Epub 2024 Sep 19.
Retinitis pigmentosa (RP) is a complex spectrum of inherited retinal diseases marked by the gradual loss of photoreceptor cells, ultimately leading to blindness. Among these, mutations in PDE6A, responsible for encoding a cGMP-specific phosphodiesterase, stand out as pivotal in autosomal recessive RP (RP43). Unfortunately, no effective therapy currently exists for this specific form of RP. However, recent advancements in genome editing, such as base editing (BE) and prime editing (PE), offer a promising avenue for precise and efficient gene therapy. Here, it is illustrated that the engineered BE and PE systems, particularly PE, exhibit high efficiency in rescuing a target point mutation with minimal bystander effects in an RP mouse model carrying the Pde6a (c.2009A > G, p.D670G) mutation. The optimized BE and PE systems are first screened in N2a cells and subsequently assessed in electroporated mouse retinas. Notably, the optimal PE system, delivered via dual adeno-associated virus (AAV), precisely corrects the pathogenic mutation with average 9.4% efficiency, with no detectable bystander editing. This correction restores PDE6A protein expression, preserved photoreceptors, and rescued retinal function in Pde6a mice. Therefore, this study offers a proof-of-concept demonstration for the treatment of Pde6a-related retinal degeneration using BE and PE systems.
色素性视网膜炎(RP)是一组复杂的遗传性视网膜疾病,其特征是感光细胞逐渐丧失,最终导致失明。在这些疾病中,突变 PDE6A 编码 cGMP 特异性磷酸二酯酶,在常染色体隐性遗传 RP(RP43)中尤为突出。不幸的是,目前针对这种特定形式的 RP 尚无有效的治疗方法。然而,最近基因组编辑的进展,如碱基编辑(BE)和先导编辑(PE),为精确和高效的基因治疗提供了有前途的途径。在这里,我们展示了工程化的 BE 和 PE 系统,特别是 PE,在携带 Pde6a(c.2009A > G,p.D670G)突变的 RP 小鼠模型中,具有高效修复目标点突变的能力,同时最小化旁观者效应。优化的 BE 和 PE 系统首先在 N2a 细胞中进行筛选,然后在电穿孔的小鼠视网膜中进行评估。值得注意的是,通过双腺相关病毒(AAV)递送的最佳 PE 系统以平均 9.4%的效率精确校正致病突变,没有可检测到的旁观者编辑。这种校正恢复了 PDE6A 蛋白的表达,保留了感光细胞,并挽救了 Pde6a 小鼠的视网膜功能。因此,本研究为使用 BE 和 PE 系统治疗 Pde6a 相关的视网膜变性提供了概念验证。