Corneal fibrosis and neovascularization (CNV) after ocular trauma impairs vision. This study tested therapeutic potential of tissue-targeted adeno-associated virus5 (AAV5) mediated decorin (DCN) and pigment epithelium-derived factor (PEDF) combination genes . Corneal fibrosis and CNV were induced in New Zealand White rabbits via chemical trauma. Gene therapy in stroma was delivered 30-min after chemical-trauma via topical AAV5-DCN and AAV5-PEDF application using a cloning cylinder. Clinical eye examinations and multimodal imaging in live rabbits were performed periodically and corneal tissues were collected 9-day and 15-day post euthanasia. Histological, cellular, and molecular and apoptosis assays were used for efficacy, tolerability, and mechanistic studies. The AAV5-DCN and AAV5-PEDF combination gene therapy significantly reduced corneal fibrosis (p < 0.01 or p < 0.001) and CNV (p < 0.001) in therapy-given (chemical-trauma and AAV5-DCN + AAV5-PEDF) rabbit eyes compared to the no-therapy given eyes (chemical-trauma and AAV5-naked vector). Histopathological analyses demonstrated significantly reduced fibrotic α-smooth muscle actin and endothelial lectin expression in therapy-given corneas compared to no-therapy corneas on day-9 (p < 0.001) and day-15 (p < 0.001). Further, therapy-given corneas showed significantly increased Fas-ligand mRNA levels (p < 0.001) and apoptotic cell death in neovessels (p < 0.001) compared to no-therapy corneas. AAV5 delivered 2.69 × 10 copies of DCN and 2.31 × 10 copies of PEDF genes per μg of DNA. AAV5 vector and delivered DCN and PEDF genes found tolerable to the rabbit eyes and caused no significant toxicity to the cornea. The combination AAV5-DCN and AAV5-PEDF topical gene therapy effectively reduces corneal fibrosis and CNV with high tolerability in rabbits. Additional studies are warranted.

中文翻译：

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组织靶向和局部 AAV5-DCN 和 AAV5-PEDF 联合基因治疗消除兔眼体内角膜纤维化和并发新生血管形成

眼外伤后的角膜纤维化和新生血管（CNV）会损害视力。本研究测试了组织靶向腺相关病毒 5 (AAV5) 介导的核心蛋白聚糖 (DCN) 和色素上皮衍生因子 (PEDF) 组合基因的治疗潜力。通过化学创伤在新西兰白兔中诱导角膜纤维化和 CNV。化学创伤后 30 分钟，使用克隆圆筒通过局部 AAV5-DCN 和 AAV5-PEDF 应用进行基质基因治疗。定期对活兔进行临床眼部检查和多模态成像，并在安乐死后 9 天和 15 天收集角膜组织。组织学、细胞、分子和细胞凋亡测定用于功效、耐受性和机制研究。 AAV5-DCN 和 AAV5-PEDF 联合基因治疗显着减少接受治疗（化学创伤和 AAV5-DCN + AAV5-PEDF）兔眼的角膜纤维化（p < 0.01 或 p < 0.001）和 CNV（p < 0.001）与未接受治疗的眼睛（化学创伤和 AAV5 裸载体）相比。组织病理学分析表明，与第 9 天 (p < 0.001) 和第 15 天 (p < 0.001) 未治疗的角膜相比，接受治疗的角膜中纤维化 α-平滑肌肌动蛋白和内皮凝集素的表达显着降低。此外，与未接受治疗的角膜相比，接受治疗的角膜显示 Fas 配体 mRNA 水平显着增加 (p < 0.001)，新生血管中的细胞凋亡显着增加 (p < 0.001)。 AAV5 每 μg DNA 传递 2.69 × 10 个拷贝的 DCN 和 2.31 × 10 个拷贝的 PEDF 基因。 AAV5 载体和递送的 DCN 和 PEDF 基因被发现对兔眼是耐受的，并且不会对角膜造成明显的毒性。 AAV5-DCN 和 AAV5-PEDF 联合局部基因疗法可有效减少兔角膜纤维化和 CNV，且具有高耐受性。需要进行额外的研究。