Zhao Guoli, Li Zhen, Zhao Ming-Jie, Li Shu-Ying, Xia Qing, Xu Shuoyu, Zhang Yu, Wang Yi, Li Fang, Liu Yu-Ling, Guo Yun-Hui, Xu Ruo-Xi, Zhou Han, Zhou Hong, Ding Wen-Wen, Wang Yong-Chen, Miao Yanying, Wang Zhongfeng
State Key Laboratory of Brain Function and Disorders and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai, 200032, China.
Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University; Key Laboratory of Myopia and Related Eye Diseases, NHC; Key Laboratory of Myopia and Related Eye Diseases, Chinese Academy of Medical Sciences, Shanghai, 200031, China.
Adv Sci (Weinh). 2025 Sep;12(33):e15856. doi: 10.1002/advs.202415856. Epub 2025 Jun 19.
Glaucoma is a neurodegenerative disease characterized by the progressive degeneration of retinal ganglion cells (RGCs) and their axons, ultimately leading to irreversible vision loss. Elevated intraocular pressure (IOP) is one of the significant risk factors in glaucoma; however, neurodegeneration continues even after effective IOP management, underscoring the need for neuroprotective therapies. This study investigates the role of connexin43 (Cx43), which is extensively expressed in retinal macroglia, in regulating microglial activation and optic nerve degeneration in glaucoma. A high-fidelity CRISPR-Cas13 (hfCas13X) system is employed to selectively target and knock down Cx43 expression in macroglia. The findings reveal that Cx43-mediated ATP release through hemichannels exacerbates microglial activation and neuroinflammation, thereby contributing to RGC loss. Notably, in a mouse model of chronic ocular hypertension (COH) glaucoma, knocking down Cx43 in macroglia using the hfCas13X system significantly promoted the survival of RGCs and the integrity of the optic nerve, and improved visual function. The hfCas13X system, which offers high-fidelity RNA editing with minimal off-target effects, represents a novel and promising therapeutic strategy for glaucoma, highlighting the potential of gene editing technologies in the management of neurodegenerative diseases.
青光眼是一种神经退行性疾病，其特征是视网膜神经节细胞（RGCs）及其轴突进行性退化，最终导致不可逆转的视力丧失。眼压升高（IOP）是青光眼的重要危险因素之一；然而，即使在有效控制眼压后，神经退行性变仍会继续，这凸显了神经保护疗法的必要性。本研究调查了在视网膜大胶质细胞中广泛表达的连接蛋白43（Cx43）在调节青光眼小胶质细胞活化和视神经退变中的作用。采用高保真CRISPR-Cas13（hfCas13X）系统选择性靶向并敲低大胶质细胞中Cx43的表达。研究结果表明，Cx43通过半通道介导的ATP释放会加剧小胶质细胞活化和神经炎症，从而导致RGCs丢失。值得注意的是，在慢性高眼压（COH）青光眼小鼠模型中，使用hfCas13X系统敲低大胶质细胞中的Cx43可显著促进RGCs的存活和视神经的完整性，并改善视觉功能。hfCas13X系统具有高保真RNA编辑且脱靶效应最小的特点，是一种新颖且有前景的青光眼治疗策略，凸显了基因编辑技术在神经退行性疾病治疗中的潜力。
