State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, 510060, China.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, 510060, China.
Exp Eye Res. 2024 Jul;244:109931. doi: 10.1016/j.exer.2024.109931. Epub 2024 May 17.
Gene therapy is one of the strategies that may reduce or reverse progressive neurodegeneration in retinal neurodegenerative diseases. However, efficiently delivering transgenes to retinal ganglion cells (RGCs) remains hard to achieve. In this study, we innovatively investigated transduction efficiency of adeno-associated virus (AAV)-PHP.eB in murine RGCs by retro-orbital venous sinus injection. Five doses of AAV-PHP.eB-EGFP were retro-orbitally injected in venous sinus in adult C57/BL6J mice. Two weeks after administration, RGCs transduction efficiency was quantified by retinal flat-mounts and frozen section co-labeling with RGCs marker Rbpms. In addition, safety of this method was evaluated by RGCs survival rate and retinal morphology. To conform efficacy of this new method, AAV-PHP.eB-CNTF was administrated into mature mice through single retro-orbital venous injection after optic nerve crush injury to evaluate axonal elongation. Results indicated that AAV- PHP.eB readily crossed the blood-retina barrier and was able to transduce more than 90% of RGCs when total dose of virus reached 5 × 10 vector genomes (vg). Moreover, this technique did not affect RGCs survival rate and retinal morphology. Furthermore, retro-orbital venous delivery of AAV-PHP.eB-CNTF effectively transduced RGCs, robustly promoted axonal regeneration after optic nerve crush injury. Thus, novel AAV-PHP.eB retro-orbital injection provides a minimally invasive and efficient route for transgene delivery in treatment of retinal neurodegenerative diseases.
基因治疗是一种可能减少或逆转视网膜神经退行性疾病进行性神经退行性变的策略。然而，将转基因有效地递送到视网膜神经节细胞（RGC）仍然难以实现。在这项研究中，我们通过眶后静脉窦内注射创新地研究了腺相关病毒（AAV）-PHP.eB 在小鼠 RGC 中的转导效率。在成年 C57/BL6J 小鼠的眶后静脉窦内注射五剂 AAV-PHP.eB-EGFP。给药后两周，通过视网膜平铺和与 RGC 标志物 Rbpms 共标记的冷冻切片定量评估 RGC 转导效率。此外，通过 RGC 存活率和视网膜形态评估该方法的安全性。为了证实这种新方法的功效，通过单次眶后静脉注射 AAV-PHP.eB-CNTF 在成熟小鼠视神经挤压损伤后进行给药，以评估轴突伸长。结果表明，当病毒总量达到 5×10 个载体基因组（vg）时，AAV-PHP.eB 容易穿过血视网膜屏障，并能够转导超过 90%的 RGC。此外，该技术不会影响 RGC 存活率和视网膜形态。此外，AAV-PHP.eB 通过眶后静脉递送有效地转导了 RGC，在视神经挤压损伤后强烈促进了轴突再生。因此，新型 AAV-PHP.eB 眶后注射为治疗视网膜神经退行性疾病的转基因递送提供了一种微创且高效的途径。
