Gene Correction Reverses Ciliopathy and Photoreceptor Loss in iPSC-Derived Retinal Organoids from Retinitis Pigmentosa Patients.
Gene Correction Reverses Ciliopathy and Photoreceptor Loss in iPSC-Derived Retinal Organoids from Retinitis Pigmentosa Patients.
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基因校正可逆转色素性视网膜炎患者 iPSC 来源的视网膜类器官的纤毛病变和光感受器损失
DOI：
10.1016/j.stemcr.2018.02.003
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发表时间：
2018-04-10
影响因子：
5.9
通讯作者：
Jin ZB
中科院分区：
文献类型：
--
作者：
Deng WL;Gao ML;Lei XL;Lv JN;Zhao H;He KW;Xia XX;Li LY;Chen YC;Li YP;Pan D;Xue T;Jin ZB
视网膜色素变性（RP）是一种不可逆的遗传性视网膜病变，其中观察到早发性夜盲症。尽管RP的遗传异质性，RPGR突变是这种疾病的最常见原因。在这里，我们从三名RPGR基因具有不同移码突变的RP患者中产生了诱导多能干细胞（iPSC），然后将其分化为视网膜色素上皮（RPE）细胞和具有电生理特性的结构良好的视网膜类器官。我们观察到光感受器在形态、定位、转录谱和电生理活性方面的显著缺陷。此外，在患者iPSC、RPE细胞和三维视网膜类器官中发现了短纤毛。CRISPR-Cas9介导的RPGR突变的校正挽救了光感受器结构和电生理特性，逆转了观察到的纤毛病变，并使用基于转录组的分析将基因表达恢复到与对照组一致的水平。这项研究使用患者特异性类器官概括了RPGR的发病机制，并在培养皿中实现了RPGR突变的靶向基因治疗作为概念验证证据。具有外节和电生理特性的HiPSC衍生的3D视网膜RPGR突变导致患者iPSC衍生的3D视网膜中的感光细胞患病突变校正挽救感光细胞形态和电生理学缺陷纤毛发生缺陷出现在RPGR患者特异性iPSC，iPSC-RPE，和3D视网膜Jin及其同事证明，患者特异性iPSC-衍生的3D视网膜可以通过呈现感光细胞形态、基因谱和电生理学的缺陷以及iPSC、iPSC-RPE和3D视网膜中有缺陷的纤毛发生来重现视网膜色素变性的疾病进展。CRISPR/Cas9介导的基因校正不仅可以挽救光感受器结构和电生理特性，还可以挽救观察到的纤毛病变。
Retinitis pigmentosa (RP) is an irreversible, inherited retinopathy in which early-onset nyctalopia is observed. Despite the genetic heterogeneity of RP, RPGR mutations are the most common causes of this disease. Here, we generated induced pluripotent stem cells (iPSCs) from three RP patients with different frameshift mutations in the RPGR gene, which were then differentiated into retinal pigment epithelium (RPE) cells and well-structured retinal organoids possessing electrophysiological properties. We observed significant defects in photoreceptor in terms of morphology, localization, transcriptional profiling, and electrophysiological activity. Furthermore, shorted cilium was found in patient iPSCs, RPE cells, and three-dimensional retinal organoids. CRISPR-Cas9-mediated correction of RPGR mutation rescued photoreceptor structure and electrophysiological property, reversed the observed ciliopathy, and restored gene expression to a level in accordance with that in the control using transcriptome-based analysis. This study recapitulated the pathogenesis of RPGR using patient-specific organoids and achieved targeted gene therapy of RPGR mutations in a dish as proof-of-concept evidence. HiPSC-derived 3D retinae with outer segments and electrophysiological properties RPGR mutation results in diseased photoreceptor in patient iPSC-derived 3D retinae Mutation correction rescues defects in photoreceptor morphology and electrophysiology Ciliogenesis defects appear in RPGR patient-specific iPSCs, iPSC-RPE, and 3D retinae Jin and colleagues demonstrate that patient-specific iPSC-derived 3D retinae can recapitulate disease progress of retinitis pigmentosa through presenting defects in photoreceptor morphology, gene profile, and electrophysiology, as well as the defective ciliogenesis in iPSCs, iPSC-RPE, and 3D retinae. CRISPR/Cas9-mediated gene correction can rescue not only photoreceptor structure and electrophysiological property but also observed ciliopathy.
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DOI：
10.1002/stem.1372
发表时间：
2013-06
期刊：
Stem cells (Dayton, Ohio)
影响因子：
--
作者：
Homma K;Okamoto S;Mandai M;Gotoh N;Rajasimha HK;Chang YS;Chen S;Li W;Cogliati T;Swaroop A;Takahashi M
通讯作者：
Takahashi M
影响因子：
3.7
作者：
Jin ZB;Okamoto S;Osakada F;Homma K;Assawachananont J;Hirami Y;Iwata T;Takahashi M
通讯作者：
Takahashi M
下一代测序揭示了一大群遗传性视网膜营养不良患者的基因型-表型相关性和突变谱
影响因子：
8.8
作者：
Huang, Xiu-Feng;Huang, Fang;Jin, Zi-Bing
通讯作者：
Jin, Zi-Bing
影响因子：
64.8
作者：
Eiraku, Mototsugu;Takata, Nozomu;Sasai, Yoshiki
通讯作者：
Sasai, Yoshiki
影响因子：
3.4
作者：
Megaw RD;Soares DC;Wright AF
通讯作者：
Wright AF