Allele-specific gene editing to rescue dominant CRX-associated LCA7 phenotypes in a retinal organoid model.
Allele-specific gene editing to rescue dominant CRX-associated LCA7 phenotypes in a retinal organoid model.
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DOI：
10.1016/j.stemcr.2021.09.007
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发表时间：
2021-11-09
影响因子：
5.9
通讯作者：
Lamba DA
中科院分区：
文献类型：
--
作者：
Chirco KR;Chew S;Moore AT;Duncan JL;Lamba DA
由CRX(LCA7)突变引起的Leber先天性黑色素症病例表现为疾病的早期形式，并显示出显着的光感受器功能障碍和最终丧失的迹象。为了建立一个体外翻译的模型系统来研究基于基因编辑的治疗方法，我们产生了含有CRX显性致病突变的LCA7视网膜有机体。我们的LCA7视网膜有机体发展出未成熟和功能障碍的光感受器细胞的迹象，为我们提供了一个可靠的体外模型来概括LCA7。此外，我们进行了一项概念验证研究，在该研究中，我们利用基于等位基因特异性CRISPR/Cas9的基因编辑来敲除突变的CRX，并看到我们的有机物中的光感受器表型得到了适度的挽救。这项工作为治疗LCA7的有效方法提供了早期证据，该方法可以更广泛地应用于其他显性遗传病。一个视网膜器官模型的CRX相关的Leber先天性黑素瘤是产生的LCA7视网膜器官显示受损的光感受器成熟和基因表达scRNA-seq图谱显示视杆和视锥感光细胞功能障碍等位基因突变CRX的编辑促进疾病挽救视网膜器官LCA7目前还没有有效的治疗方法；因此，Lamba博士和他的同事从患者来源的干细胞产生了LCA7视网膜器官。视网膜器官重现了LCA7疾病的表型，允许测试等位基因特异的基因编辑方法。疾病抢救的早期迹象已经实现，为治疗LCA7和其他主要视网膜疾病提供了有前景的新策略的证据。
Cases of Leber congenital amaurosis caused by mutations in CRX (LCA7) exhibit an early form of the disease and show signs of significant photoreceptor dysfunction and eventual loss. To establish a translational in vitro model system to study gene-editing-based therapies, we generated LCA7 retinal organoids harboring a dominant disease-causing mutation in CRX. Our LCA7 retinal organoids develop signs of immature and dysfunctional photoreceptor cells, providing us with a reliable in vitro model to recapitulate LCA7. Furthermore, we performed a proof-of-concept study in which we utilize allele-specific CRISPR/Cas9-based gene editing to knock out mutant CRX and saw moderate rescue of photoreceptor phenotypes in our organoids. This work provides early evidence for an effective approach to treat LCA7, which can be applied more broadly to other dominant genetic diseases. A retinal organoid model of CRX-associated Leber congenital amaurosis was generated LCA7 retinal organoids show impaired photoreceptor maturation and gene expression scRNA-seq profiling revealed both rod and cone photoreceptor dysfunction Allele-specific editing of mutant CRX promotes disease rescue in retinal organoids LCA7 currently has no effective treatments; therefore, Dr. Lamba and colleagues generated LCA7 retinal organoids from patient-derived stem cells. The retinal organoids recapitulated LCA7 disease phenotypes, allowing for testing of an allele-specific gene editing approach. Early signs of disease rescue were achieved, providing evidence for a promising new strategy to treat LCA7 and other dominant retinal diseases.
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