Optimization of Retinal Gene Therapy for X-Linked Retinitis Pigmentosa Due to RPGR Mutations
Optimization of Retinal Gene Therapy for X-Linked Retinitis Pigmentosa Due to RPGR Mutations
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DOI：
10.1016/j.ymthe.2017.05.004
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发表时间：
2017-08-02
影响因子：
12.4
通讯作者：
Aguirre, Gustavo D.
中科院分区：
文献类型：
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作者：
Beltran, William A.;Cideciyan, Artur V.;Aguirre, Gustavo D.
由 RPGR 基因突变引起的 X 连锁视网膜色素变性 (XLRP) 是一种早发且严重的失明原因。最近在犬类模型中成功的概念验证研究表明，开发 RPGR-XLRP 的纠正基因疗法现在可能是一个可以实现的目标。为了准备未来的临床试验，我们优化了治疗性 AAV 载体构建体，表明 GRK1（而不是 IRBP）是一种更有效的启动子，可以将基因表达靶向非人类灵长类动物的视杆细胞和视锥细胞。在 GRK1 启动子的控制下，在 RPGR 突变体 (XLPRA2) 狗中使用了两个转基因。第一个是之前开发的稳定的人类RPGR（hRPGRstb）。第二个是新的全长稳定和密码子优化的人类 RPGR (hRPGRco)。对携带受 GRK1 启动子控制的 hRPGRstb 的 AAV2/5 载体进行的长期（>2 年）研究表明，可以挽救视杆细胞和视锥细胞的退化并保留视力。短期（3 个月）研究表明，用携带 GRK1 启动子控制下任一转基因的 AAV2/5 载体处理的犬眼中，光感受器的保存情况相当。这些结果提供了关键的分子组件（GRK1 启动子、hRPGRco 转基因），用于构建针对 RPGR-XLRP 患者优化的治疗性病毒载体。
X-linked retinitis pigmentosa (XLRP) caused by mutations in the RPGR gene is an early onset and severe cause of blindness. Successful proof-of-concept studies in a canine model have recently shown that development of a corrective gene therapy for RPGR-XLRP may now be an attainable goal. In preparation for a future clinical trial, we have here optimized the therapeutic AAV vector construct by showing that GRK1 (rather than IRBP) is a more efficient promoter for targeting gene expression to both rods and cones in non-human primates. Two transgenes were used in RPGR mutant (XLPRA2) dogs under the control of the GRK1 promoter. First was the previously developed stabilized human RPGR (hRPGRstb). Second was a new full-length stabilized and codon-optimized human RPGR (hRPGRco). Long-term (>2 years) studies with an AAV2/5 vector carrying hRPGRstb under control of the GRK1 promoter showed rescue of rods and cones from degeneration and retention of vision. Shorter term (3 months) studies demonstrated comparable preservation of photoreceptors in canine eyes treated with an AAV2/5 vector carrying either transgene under the control of the GRK1 promoter. These results provide the critical molecular components (GRK1 promoter, hRPGRco transgene) to now construct a therapeutic viral vector optimized for RPGR-XLRP patients.