Joint Shantou International Eye Center of Shantou University and The Chinese University of Hong Kong, Shantou, Guangdong, China; Shantou University Medical College, Shantou, Guangdong, China.
Joint Shantou International Eye Center of Shantou University and The Chinese University of Hong Kong, Shantou, Guangdong, China.
Exp Eye Res. 2020 Apr;193:107955. doi: 10.1016/j.exer.2020.107955. Epub 2020 Feb 1.
Gene therapy has been proposed as a feasible strategy for RGC survival and optic nerve regeneration. Some preclinical and clinical studies revealed intraocular inflammation after intravitreal injection of adeno-associated virus (AAV) by slit-lamp or indirect ophthalmoscope. Here we evaluate the longitudinal profile of immediate inflammatory responses after AAV2 injection in rat retina and vitreous body by optical coherence tomography (OCT). Adult Fischer F344 rats were intravitreally injected once with saline, AAV2 or zymosan. Retinal thickness and cell infiltration were recorded by OCT longitudinally for 2 months and verified by histological analysis. The transduction rate of single intravitreal AAV2 injection was 21.3 ± 4.9% of whole retina, and the transduction efficiency on RGCs was 91.5 ± 2.5% in the transduced area. Significant increase in cell infiltration was observed from Day 1-3 after AAV2 injection, compared to very few infiltrating cells observed in the saline-injected group. The infiltrating cells ceased at Day 5 after intravitreal injection and remained absent at 2 months. The thicknesses of total and inner retina were increased along Day 1-3 after AAV2 injection, but reverted to normal afterwards. The surviving RGCs in the AAV2-injected groups at Day 14 showed no significant difference compared to saline-injected group. In summary, this study revealed the immediate inflammatory responses and retinal edema after intravitreal AAV2 injection in normal rats, without influencing long-term retinal thickness and RGC survival. OCT can be implemented for the time-lapse in vivo evaluation of inflammatory response after AAV-mediated gene therapy through intravitreal injection.
基因治疗被提出作为一种可行的策略，用于视网膜神经节细胞（RGC）的存活和视神经再生。一些临床前和临床研究显示，腺相关病毒（AAV）经玻璃体内注射后，通过裂隙灯或间接检眼镜观察到眼内炎症。在这里，我们通过光学相干断层扫描（OCT）评估 AAV2 注射后大鼠视网膜和玻璃体的即刻炎症反应的纵向特征。成年 Fischer F344 大鼠经玻璃体内单次注射盐水、AAV2 或酵母聚糖。通过 OCT 纵向记录视网膜厚度和细胞浸润 2 个月，并通过组织学分析进行验证。单次玻璃体内 AAV2 注射的转导率为整个视网膜的 21.3±4.9%，转导区域的 RGC 转导效率为 91.5±2.5%。与盐水注射组观察到的极少数浸润细胞相比，AAV2 注射后第 1-3 天观察到细胞浸润显著增加。玻璃体内注射后第 5 天，浸润细胞停止，2 个月后仍未出现。AAV2 注射后第 1-3 天，全层和内层视网膜厚度增加，但随后恢复正常。AAV2 注射组在第 14 天的存活 RGC 与盐水注射组相比无显著差异。总之，本研究揭示了正常大鼠玻璃体内注射 AAV2 后的即刻炎症反应和视网膜水肿，而不影响长期视网膜厚度和 RGC 存活。OCT 可用于通过玻璃体内注射实现 AAV 介导的基因治疗后炎症反应的实时体内评估。
