Genome Editing <i>VEGFA</i> Prevents Corneal Neovascularization In Vivo
Zhenhai Zeng, Siheng Li, Xiuhong Ye, Yiran Wang, Qinmei Wang, Zhongxing Chen, Ziqian Wang, Jun Zhang, Qing Wang, Lu Chen, Shuangzhe Zhang, Zhilin Zou, Meimin Lin, Xinyi Chen, Guoli Zhao, Colm McAlinden, Hetian Lei, Xingtao Zhou, Jinhai Huang
Abstract
Corneal neovascularization (CNV) is a common clinical finding seen in a range of eye diseases. Current therapeutic approaches to treat corneal angiogenesis, in which vascular endothelial growth factor (VEGF) A plays a central role, can cause a variety of adverse side effects. The technology of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 can edit VEGFA gene to suppress its expression. CRISPR offers a novel opportunity to treat CNV. This study shows that depletion of VEGFA with a novel CRISPR/Cas9 system inhibits proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs) in vitro. Importantly, subconjunctival injection of this dual AAV-SpCas9/sgRNA-VEGFA system is demonstrated which blocks suture-induced expression of VEGFA, CD31, and α-smooth muscle actin as well as corneal neovascularization in mice. This study has established a strong foundation for the treatment of corneal neovascularization via a gene editing approach for the first time.