Huang Xionggao, Zhou Guohong, Wu Wenyi, Duan Yajian, Ma Gaoen, Song Jingyuan, Xiao Ru, Vandenberghe Luk, Zhang Feng, D'Amore Patricia A, Lei Hetian
Schepens Eye Research Institute of Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, 02114, USA.
Department of Ophthalmology, Harvard Medical School, Boston, MA, 02114, USA.
Nat Commun. 2017 Jul 24;8(1):112. doi: 10.1038/s41467-017-00140-3.
Angiogenesis, in which vascular endothelial growth factor receptor (VEGFR) 2 plays an essential role, is associated with a variety of human diseases including proliferative diabetic retinopathy and wet age-related macular degeneration. Here we report that a system of adeno-associated virus (AAV)-mediated clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease (Cas)9 from Streptococcus pyogenes (SpCas9) is used to deplete VEGFR2 in vascular endothelial cells (ECs), whereby the expression of SpCas9 is driven by an endothelial-specific promoter of intercellular adhesion molecule 2. We further show that recombinant AAV serotype 1 (rAAV1) transduces ECs of pathologic vessels, and that editing of genomic VEGFR2 locus using rAAV1-mediated CRISPR/Cas9 abrogates angiogenesis in the mouse models of oxygen-induced retinopathy and laser-induced choroid neovascularization. This work establishes a strong foundation for genome editing as a strategy to treat angiogenesis-associated diseases.Abnormal angiogenesis causes many ocular diseases. Here the authors employ CRISPR/Cas9 gene editing technology to silence VEGFR2, a major regulator of angiogenesis, in retinal endothelium and abrogate angiogenesis in the mouse models of oxygen-induced retinopathy and laser-induced choroid neovascularization.
血管生成与多种人类疾病相关，包括增殖性糖尿病视网膜病变和湿性年龄相关性黄斑变性，其中血管内皮生长因子受体（VEGFR）2起着至关重要的作用。在此，我们报告了一种腺相关病毒（AAV）介导的来自化脓性链球菌（SpCas9）的成簇规律间隔短回文重复序列（CRISPR）相关核酸酶（Cas）9系统，该系统用于耗尽血管内皮细胞（ECs）中的VEGFR2，其中SpCas9的表达由细胞间粘附分子2的内皮特异性启动子驱动。我们进一步表明，重组1型腺相关病毒（rAAV1）可转导病理性血管的内皮细胞，并且使用rAAV1介导的CRISPR/Cas9对基因组VEGFR2位点进行编辑可消除氧诱导性视网膜病变和激光诱导性脉络膜新生血管小鼠模型中的血管生成。这项工作为将基因组编辑作为治疗血管生成相关疾病策略奠定了坚实基础。异常血管生成会导致许多眼部疾病。在此，作者采用CRISPR/Cas9基因编辑技术使血管生成的主要调节因子VEGFR2在视网膜内皮中沉默，并消除氧诱导性视网膜病变和激光诱导性脉络膜新生血管小鼠模型中的血管生成。
