Nme(2) Cas9-mediated therapeutic editing in inhibiting angiogenesis after wet age-related macular degeneration onset.
Nme(2) Cas9-mediated therapeutic editing in inhibiting angiogenesis after wet age-related macular degeneration onset.
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DOI：
10.1002/ctm2.1383
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发表时间：
2023-08
影响因子：
10.6
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年龄相关性黄斑变性（AMD），特别是以脉络膜新生血管（CNV）为特征的湿性黄斑变性，是老年人视力丧失的主要原因。缺氧诱导因子-1α (HIF-1α)/血管内皮生长因子 (VEGF)/VEGF 受体 2 (VEGFR2) 通路参与 CNV 发病机制。之前的基因编辑研究表明，破坏视网膜色素上皮细胞中的这些基因可以对 CNV 进展起到预防作用。然而，目前尚未开展使用基因编辑在 CNV 诱导后破坏 VEGF 信号传导进行治疗验证的研究，这对于湿性 AMD 基因编辑疗法的临床应用至关重要。在这里，我们在诱导 CNV 后使用单腺相关病毒介导的 Nme2Cas9 来破坏 VEGF 信号传导中的关键分子 Hif1α、Vegfa 和 Vegfr2，并评估其治疗效果。我们发现 Nme2Cas9 在体内 11 天后对目标基因的编辑效率高达 71.8%。与阴性对照相比，在CNV发展早期仅使用Nme2Cas9-Vegfa治疗可使CNV病变面积减少49.5%，而Nme2Cas9-Hif1α或Nme2Cas9-Vegfr2治疗没有显示出治疗效果。此外，在体内 Nme2Cas9 介导的基因编辑中没有观察到脱靶效应。这项研究提供了利用 Nme2Cas9 进行湿性 AMD 潜在抗血管生成治疗的概念验证可能性。单个 AAV 传递的 Nme2Cas9 可在短时间内介导体内湿性 AMD 相关基因的有效编辑。靶向Vegfa基因的Nme2Cas9在湿性AMD发病后通过早期干预显着减少了CNV病变面积49.5%，而靶向Hif1α或Vegfr2的Nme2Cas9没有显示出显着的治疗效果。 Nme2Cas9 介导的体内基因编辑未观察到脱靶效应。
Age‐related macular degeneration (AMD), particularly wet AMD characterised by choroidal neovascularization (CNV), is a leading cause of vision loss in the elderly. The hypoxia‐inducible factor‐1α (HIF‐1α)/vascular endothelial growth factor (VEGF)/VEGF receptor 2 (VEGFR2) pathway contributes to CNV pathogenesis. Previous gene editing research indicated that disrupting these genes in retinal pigment epithelial cells could have a preventive effect on CNV progression. However, no studies have yet been conducted using gene editing to disrupt VEGF signalling after CNV induction for therapeutic validation, which is critical to the clinical application of wet AMD gene editing therapies. Here, we employed the single‐adeno‐associated virus‐mediated Nme2Cas9 to disrupt key molecules in VEGF signalling, Hif1α, Vegfa and Vegfr2 after inducing CNV and estimated their therapeutic effects. We found that Nme2Cas9 made efficient editing in target genes up to 71.8% post 11 days in vivo. And only Nme2Cas9‐Vegfa treatment during the early stage of CNV development reduced the CNV lesion area by 49.5%, compared to the negative control, while Nme2Cas9‐Hif1α or Nme2Cas9‐Vegfr2 treatment did not show therapeutic effect. Besides, no off‐target effects were observed in Nme2Cas9‐mediated gene editing in vivo. This study provides proof‐of‐concept possibility of employing Nme2Cas9 for potential anti‐angiogenesis therapy in wet AMD. Single AAV‐delivered Nme2Cas9 mediated efficient editing of wet AMD‐associated genes in vivo in a short time. Nme2Cas9 targeting Vegfa gene significantly reduced the CNV lesion area by 49.5% after wet AMD onset with an early‐stage intervention, while Nme2Cas9 targeting Hif1α or Vegfr2 did not show significant therapeutic effect. No off‐target effects were observed in Nme2Cas9‐mediated gene editing in vivo.
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影响因子：
46.9
作者：
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