Cone-Specific Promoters for Gene Therapy of Achromatopsia and Other Retinal Diseases
Cone-Specific Promoters for Gene Therapy of Achromatopsia and Other Retinal Diseases
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DOI：
10.1089/hum.2015.130
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发表时间：
2016-01-01
影响因子：
4.2
通讯作者：
Chulay, Jeffrey D.
中科院分区：
文献类型：
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作者：
Ye, Guo-Jie;Budzynski, Ewa;Chulay, Jeffrey D.
含有视锥细胞特异性启动子的腺相关病毒（AAV）载体在色盲的小鼠和狗模型中挽救了视锥细胞感光器功能，但视锥细胞特异性启动子尚未优化用于灵长类动物。使用通过视网膜下注射施用的AAV载体，我们评估了基于人L-视蛋白启动子或嵌合人视锥细胞转导蛋白启动子的一系列启动子在小鼠和非人灵长类动物中驱动绿色荧光蛋白（GFP）基因表达的能力。这些启动子中的每一个都指导小鼠光感受器中的高水平GFP表达。在灵长类动物中，视网膜下注射含有1.7-kb L-视蛋白启动子（PR1.7）的AAV-GFP载体在所有视锥光感受器中实现了强烈和特异性的GFP表达，并且比在拯救色盲小鼠和狗模型中的视锥功能的AAV载体中使用的含有2.1-kb L-视蛋白启动子的载体更有效。在小鼠和狗视锥光感受器中指导强GFP表达的嵌合视锥转导蛋白启动子不能驱动灵长类视锥中的GFP表达。表达由PR1.7启动子驱动的人CNGB 3基因的AAV载体在色盲小鼠模型中拯救视锥细胞功能。这些结果为用于治疗色盲患者的AAV载体的设计提供了信息。
Adeno-associated viral (AAV) vectors containing cone-specific promoters have rescued cone photoreceptor function in mouse and dog models of achromatopsia, but cone-specific promoters have not been optimized for use in primates. Using AAV vectors administered by subretinal injection, we evaluated a series of promoters based on the human L-opsin promoter, or a chimeric human cone transducin promoter, for their ability to drive gene expression of green fluorescent protein (GFP) in mice and nonhuman primates. Each of these promoters directed high-level GFP expression in mouse photoreceptors. In primates, subretinal injection of an AAV-GFP vector containing a 1.7-kb L-opsin promoter (PR1.7) achieved strong and specific GFP expression in all cone photoreceptors and was more efficient than a vector containing the 2.1-kb L-opsin promoter that was used in AAV vectors that rescued cone function in mouse and dog models of achromatopsia. A chimeric cone transducin promoter that directed strong GFP expression in mouse and dog cone photoreceptors was unable to drive GFP expression in primate cones. An AAV vector expressing a human CNGB3 gene driven by the PR1.7 promoter rescued cone function in the mouse model of achromatopsia. These results have informed the design of an AAV vector for treatment of patients with achromatopsia.