Nme Cas9 介导的治疗性编辑抑制湿性年龄相关性黄斑变性发病后的血管生成。

Nme Cas9-mediated therapeutic editing in inhibiting angiogenesis after wet age-related macular degeneration onset.

机构信息

MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.

The State Key Laboratory of Ophthalmology Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.

出版信息

Clin Transl Med. 2023 Aug;13(8):e1383. doi: 10.1002/ctm2.1383.

DOI:10.1002/ctm2.1383

PMID:37598400

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10440058/

Abstract

BACKGROUND

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.

METHOD

Here, we employed the single-adeno-associated virus-mediated Nme Cas9 to disrupt key molecules in VEGF signalling, Hif1α, Vegfa and Vegfr2 after inducing CNV and estimated their therapeutic effects.

RESULTS

We found that Nme Cas9 made efficient editing in target genes up to 71.8% post 11 days in vivo. And only Nme Cas9-Vegfa treatment during the early stage of CNV development reduced the CNV lesion area by 49.5%, compared to the negative control, while Nme Cas9-Hif1α or Nme Cas9-Vegfr2 treatment did not show therapeutic effect. Besides, no off-target effects were observed in Nme Cas9-mediated gene editing in vivo.

CONCLUSIONS

This study provides proof-of-concept possibility of employing Nme Cas9 for potential anti-angiogenesis therapy in wet AMD.

摘要

背景

年龄相关性黄斑变性（AMD），特别是伴有脉络膜新生血管（CNV）的湿性 AMD，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）/血管内皮生长因子受体 2（VEGFR2）通路有助于 CNV 的发病机制。先前的基因编辑研究表明，在视网膜色素上皮细胞中破坏这些基因可能对 CNV 的进展具有预防作用。然而，迄今为止，尚未有研究使用基因编辑在 CNV 诱导后破坏 VEGF 信号来进行治疗验证，这对于湿性 AMD 基因编辑治疗的临床应用至关重要。

方法

在这里，我们采用单腺相关病毒介导的 Nme Cas9 破坏 VEGF 信号通路中的关键分子 Hif1α、Vegfa 和 Vegfr2，在诱导 CNV 后评估它们的治疗效果。

结果

我们发现，Nme Cas9 在体内第 11 天的靶基因编辑效率高达 71.8%。只有在 CNV 发展的早期阶段用 Nme Cas9-Vegfa 治疗，才能使 CNV 病变面积减少 49.5%，与阴性对照组相比，而 Nme Cas9-Hif1α 或 Nme Cas9-Vegfr2 治疗则没有治疗效果。此外，在体内 Nme Cas9 介导的基因编辑中没有观察到脱靶效应。