Telethon Institute of Genetics and Medicine, 80078 Napoli, Italy.
Department of Translational Medicine, University of Naples Federico II, 80131 Naples, Italy.
Int J Mol Sci. 2021 May 28;22(11):5767. doi: 10.3390/ijms22115767.
Gene-expression programs modulated by transcription factors (TFs) mediate key developmental events. Here, we show that the synthetic transcriptional repressor (TR; ZF6-DB), designed to treat Rhodopsin-mediated autosomal dominant retinitis pigmentosa (RHO-adRP), does not perturb murine retinal development, while maintaining its ability to block Rho expression transcriptionally. To express ZF6-DB into the developing retina, we pursued two approaches, (i) the retinal delivery (somatic expression) of ZF6-DB by Adeno-associated virus (AAV) vector (AAV-ZF6-DB) gene transfer during retinogenesis and (ii) the generation of a transgenic mouse (germ-line transmission, TR-ZF6-DB). Somatic and transgenic expression of ZF6-DB during retinogenesis does not affect retinal function of wild-type mice. The P347S mouse model of RHO-adRP, subretinally injected with AAV-ZF6-DB, or crossed with TR-ZF6-DB or shows retinal morphological and functional recovery. We propose the use of developmental transitions as an effective mode to challenge the safety of retinal gene therapies operating at genome, transcriptional, and transcript levels.
转录因子 (TFs) 调节的基因表达程序介导关键的发育事件。在这里，我们表明，设计用于治疗视蛋白介导的常染色体显性视网膜色素变性 (RHO-adRP) 的合成转录抑制剂 (TR; ZF6-DB) 不会干扰小鼠视网膜的发育，同时保持其转录阻断 Rho 表达的能力。为了将 ZF6-DB 表达到发育中的视网膜中，我们采用了两种方法，（i）在视网膜发生期间通过腺相关病毒 (AAV) 载体（AAV-ZF6-DB）基因转移进行视网膜内递送（体细胞表达）ZF6-DB，和（ii）生成转基因小鼠（种系传递，TR-ZF6-DB）。在视网膜发生期间，ZF6-DB 的体细胞和转基因表达不会影响野生型小鼠的视网膜功能。用 AAV-ZF6-DB 亚视网膜注射或与 TR-ZF6-DB 杂交的 P347S RHO-adRP 小鼠模型显示视网膜形态和功能恢复。我们提出将发育转变用作挑战在基因组、转录和转录本水平上运作的视网膜基因治疗安全性的有效模式。
