Development of a gene-editing approach to restore vision loss in Leber congenital amaurosis type 10
Development of a gene-editing approach to restore vision loss in Leber congenital amaurosis type 10
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DOI：
10.1038/s41591-018-0327-9
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发表时间：
2019-02-01
期刊：
影响因子：
82.9
通讯作者：
Jiang, Haiyan
中科院分区：
文献类型：
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作者：
Maeder, Morgan L.;Stefanidakis, Michael;Jiang, Haiyan
Leber先天性黑朦10型是由CEP290基因突变引起的严重视网膜营养不良（1,2）。我们开发了一种候选基因组编辑治疗药物EDIT-101，用于去除由CEP290基因IVS26突变产生的异常剪接供体，并恢复正常的CEP290表达。这种治疗的关键是，我们鉴定了一对金黄色葡萄球菌Cas9引导rna，它们对人类CEP290靶序列具有高度活性和特异性。在人细胞和视网膜外植体中进行的体外实验证实了其作用的分子机制和核酸酶特异性。在人源化CEP290小鼠的视网膜下递送EDIT-101显示出快速和持续的CEP290基因编辑。一种可比较的替代非人灵长类动物（NHP）载体也实现了NHP CEP290基因的高效编辑，其水平达到了目标治疗阈值，并证明了CRISPR/Cas9在体内编辑体细胞灵长类动物细胞的能力。这些结果支持进一步开发用于LCA10的EDIT-101和其他基于crispr的用于其他遗传性视网膜疾病的药物。
Leber congenital amaurosis type 10 is a severe retinal dystrophy caused by mutations in the CEP290 gene(1,2). We developed EDIT-101, a candidate genome-editing therapeutic, to remove the aberrant splice donor created by the IVS26 mutation in the CEP290 gene and restore normal CEP290 expression. Key to this therapeutic, we identified a pair of Staphylococcus aureus Cas9 guide RNAs that were highly active and specific to the human CEP290 target sequence. In vitro experiments in human cells and retinal explants demonstrated the molecular mechanism of action and nuclease specificity. Subretinal delivery of EDIT-101 in humanized CEP290 mice showed rapid and sustained CEP290 gene editing. A comparable surrogate non-human primate (NHP) vector also achieved productive editing of the NHP CEP290 gene at levels that met the target therapeutic threshold, and demonstrated the ability of CRISPR/Cas9 to edit somatic primate cells in vivo. These results support further development of EDIT-101 for LCA10 and additional CRISPR-based medicines for other inherited retinal disorders.