Askou Anne Louise, Alsing Sidsel, Benckendorff Josephine N E, Holmgaard Andreas, Mikkelsen Jacob Giehm, Aagaard Lars, Bek Toke, Corydon Thomas J
Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark.
Department of Ophthalmology, Aarhus University Hospital, 8000 Aarhus C, Denmark.
Mol Ther Nucleic Acids. 2019 Jun 7;16:38-50. doi: 10.1016/j.omtn.2019.01.012. Epub 2019 Feb 2.
Vascular endothelial growth factor A (VEGFA) is involved in the pathogenesis of vasoproliferative retinal diseases, such as exudative age-related macular degeneration (AMD). The objective of this study was to investigate whether dual-acting therapy based on the simultaneous expression of anti-VEGFA microRNAs (miRNAs) and the secreted, antiangiogenic protein pigment endothelial-derived factor (PEDF) delivered by adeno-associated virus (AAV) vectors provides improved protection against choroidal neovascularization (CNV). To investigate this, a multigenic AAV vector allowing retina pigment epithelium (RPE)-specific expression of anti-VEGFA miRNAs and PEDF was engineered. Robust expression of PEDF, driven by the RPE-specific vitelliform macular dystrophy 2 promoter, was observed in human cells and in mouse retina. A significant reduction in CNV was observed in a laser-induced CNV mouse model 57 days post-injection of the AAV5 particles conveying either anti-VEGFA miRNA and PEDF dual therapy or anti-VEGFA miRNA monotherapy. Overall, CNV reduction was most prominent in animals receiving dual-acting therapy. In both cases, the reduction in CNV was accompanied by a significant attenuation of VEGFA. In conclusion, the presented data reveal that gene therapy targeting VEGFA via multigenic AAV vectors displays combined efficacy, suggesting that dual-acting therapy is an important tool in future eye gene therapy for the treatment of neovascular ocular diseases, including AMD.
血管内皮生长因子A（VEGFA）参与血管增生性视网膜疾病的发病机制，如渗出性年龄相关性黄斑变性（AMD）。本研究的目的是调查基于腺相关病毒（AAV）载体同时表达抗VEGFA微小RNA（miRNA）和分泌型抗血管生成蛋白色素内皮衍生因子（PEDF）的双效疗法是否能更好地预防脉络膜新生血管形成（CNV）。为了对此进行研究，构建了一种多基因AAV载体，使其能在视网膜色素上皮（RPE）细胞中特异性表达抗VEGFA miRNA和PEDF。在人类细胞和小鼠视网膜中，观察到由RPE特异性卵黄状黄斑营养不良2启动子驱动的PEDF的强烈表达。在激光诱导的CNV小鼠模型中，注射携带抗VEGFA miRNA和PEDF双效疗法或抗VEGFA miRNA单效疗法的AAV5颗粒57天后，观察到CNV显著减少。总体而言，在接受双效疗法的动物中，CNV减少最为显著。在这两种情况下，CNV的减少都伴随着VEGFA的显著减弱。总之，所呈现的数据表明，通过多基因AAV载体靶向VEGFA的基因疗法具有联合疗效，这表明双效疗法是未来眼部基因治疗中治疗包括AMD在内的新生血管性眼病的重要工具。
