Du Wei, Tao Ye, Deng Wen-Tao, Zhu Ping, Li Jie, Dai Xufeng, Zhang Yuxin, Shi Wei, Liu Xuan, Chiodo Vince A, Ding Xi-Qin, Zhao Chen, Michalakis Stylianos, Biel Martin, Zhang Zuoming, Qu Jia, Hauswirth William W, Pang Ji-Jing
Deparment of Ophthalmology, University of Florida, Gainesville, FL 32610, USA.
Deparment of Ophthalmology, University of Florida, Gainesville, FL 32610, USA, School of Ophthalmology and Optometry, The Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325027, China.
Hum Mol Genet. 2015 Jul 1;24(13):3699-707. doi: 10.1093/hmg/ddv114. Epub 2015 Apr 8.
The CNGA3(-/-)/Nrl(-/-) mouse is a cone-dominant model with Cnga3 channel deficiency, which partially mimics the all cone foveal structure of human achromatopsia 2 with CNGA3 mutations. Although subretinal (SR) AAV vector administration can transfect retinal cells efficiently, the injection-induced retinal detachment can cause retinal damage, particularly when SR vector bleb includes the fovea. We therefore explored whether cone function-structure could be rescued in CNGA3(-/-)/Nrl(-/-) mice by intravitreal (IVit) delivery of tyrosine to phenylalanine (Y-F) capsid mutant AAV8. We find that AAV-mediated CNGA3 expression can restore cone function and rescue structure following IVit delivery of AAV8 (Y447, 733F) vector. Rescue was assessed by restoration of the cone-mediated electroretinogram (ERG), optomotor responses, and cone opsin immunohistochemistry. Demonstration of gene therapy in a cone-dominant mouse model by IVit delivery provides a potential alternative vector delivery mode for safely transducing foveal cones in achromatopsia patients and in other human retinal diseases affecting foveal function.
CNGA3(-/-)/Nrl(-/-)小鼠是一种具有Cnga3通道缺陷的视锥细胞主导型模型,它部分模拟了患有CNGA3突变的人类全视锥中央凹型色盲2的全视锥中央凹结构。尽管视网膜下(SR)腺相关病毒(AAV)载体给药可有效转染视网膜细胞,但注射引起的视网膜脱离会导致视网膜损伤,尤其是当SR载体泡囊包括中央凹时。因此,我们探究了通过玻璃体内(IVit)递送酪氨酸突变为苯丙氨酸(Y-F)的衣壳突变型AAV8,能否在CNGA3(-/-)/Nrl(-/-)小鼠中挽救视锥细胞功能-结构。我们发现,在玻璃体内递送AAV8(Y447, 733F)载体后,AAV介导的CNGA3表达可恢复视锥细胞功能并挽救其结构。通过视锥细胞介导的视网膜电图(ERG)、视动反应和视锥视蛋白免疫组织化学的恢复来评估挽救效果。通过玻璃体内递送在视锥细胞主导型小鼠模型中进行基因治疗的证明,为安全转导色盲患者和其他影响中央凹功能的人类视网膜疾病中的中央凹视锥细胞提供了一种潜在的替代载体递送方式。