Department of Microbiology, University of Ulsan College of Medicine, Seoul, Korea.
Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Seoul, Korea.
Invest Ophthalmol Vis Sci. 2018 Nov 1;59(13):5398-5407. doi: 10.1167/iovs.18-24926.
With anti-VEGF-based treatments for wet AMD requiring frequent injections, it is often burdensome to both patients and healthcare providers. To explore its possibility as a desirable alternative, we investigated the therapeutic potential of a recombinant adeno-associated virus 2 expressing a soluble variant of VEGF receptor-1 (rAAV2-sVEGFRv-1) in a laser-induced choroidal neovascularization (CNV) model, as CNV is a defining feature of AMD progression.
C57/B6 mice were intravitreally administered with rAAV2-sVEGFRv-1, rAAV2-GFP, or clinically used bevacizumab after CNV lesions were induced via laser photocoagulation. Immunostaining was performed with phalloidin and CD31 to measure CNV extensiveness, F4/80 and CD11b for inflammatory cell infiltration, and pan-cytokeratin to visualize fibrotic progression.
rAAV2-sVEGFRv-1 (5.0 × 107 viral genomes) possesses antiangiogenic, anti-inflammatory, and antifibrotic properties. rAAV2-sVEGFRv-1 was demonstrated to significantly decrease retinal CNV lesion size (1336 ± 186) when compared to rAAV2-GFP-treated (2949 ± 437, P = 0.0043), mock-treated (3075 ± 265, P = 0.0013), and bevacizumab-treated models (995 ± 234). Infiltration by inflammatory cells significantly decreased with rAAV2-sVEGFRv-1 administration, while groups treated with rAAV2-GFP did not. Additionally, antiapoptotic activity was observed via TUNEL assay in rAAV2-sVEGFRv-1 (16.0 ± 3.6) and rAAV2-GFP (46.0 ± 7.5, P = 0.003). Overall, the rAAV2-sVEGFRv-1 viral vector was positively comparable to bevacizumab, indicating it as effective as approved therapeutics.
The ability of a low dose of rAAV2-sVEGFRv-1 to exert a therapeutically relevant anti-VEGF effect in a CNV model is demonstrated, and strongly suggests gene therapy as an effective and convenient treatment for sustained VEGF suppression.
抗血管内皮生长因子（VEGF）药物治疗湿性年龄相关性黄斑变性（AMD）需要频繁注射，这给患者和医疗保健提供者都带来了沉重负担。为了探索其作为理想替代方案的可能性，我们研究了表达 VEGF 受体-1 可溶性变体的重组腺相关病毒 2（rAAV2-sVEGFRv-1）在激光诱导脉络膜新生血管（CNV）模型中的治疗潜力，因为 CNV 是 AMD 进展的特征。
在通过激光光凝诱导 CNV 病变后，C57/B6 小鼠通过玻璃体内注射 rAAV2-sVEGFRv-1、rAAV2-GFP 或临床使用的贝伐单抗。用鬼笔环肽和 CD31 进行免疫染色，以测量 CNV 的扩展程度，用 F4/80 和 CD11b 测量炎症细胞浸润，用泛细胞角蛋白可视化纤维化进展。
rAAV2-sVEGFRv-1（5.0×107 病毒基因组）具有抗血管生成、抗炎和抗纤维化特性。与 rAAV2-GFP 治疗组（2949±437，P=0.0043）、模拟治疗组（3075±265，P=0.0013）和贝伐单抗治疗组（995±234）相比，rAAV2-sVEGFRv-1 可显著降低视网膜 CNV 病变大小（1336±186）。rAAV2-sVEGFRv-1 给药后，炎症细胞浸润显著减少，而 rAAV2-GFP 治疗组则没有。此外，通过 TUNEL 检测，rAAV2-sVEGFRv-1 组（16.0±3.6）和 rAAV2-GFP 组（46.0±7.5，P=0.003）观察到抗细胞凋亡活性。总的来说，rAAV2-sVEGFRv-1 病毒载体与贝伐单抗的效果相当，表明其与已批准的治疗药物同样有效。
低剂量 rAAV2-sVEGFRv-1 在 CNV 模型中发挥治疗相关抗 VEGF 作用的能力得到了证明，这强烈表明基因治疗作为一种有效的、方便的持续 VEGF 抑制治疗方法。
