Human iPSC Modeling Reveals Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy.,American Journal of Human Genetics

当前位置： X-MOL 学术 › Am. J. Hum. Genet. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Human iPSC Modeling Reveals Mutation-Specific Responses to Gene Therapy in a Genotypically Diverse Dominant Maculopathy.
American Journal of Human Genetics ( IF 8.1 ) Pub Date : 2020-07-23 , DOI: 10.1016/j.ajhg.2020.06.011
Divya Sinha ₁ , Benjamin Steyer ₂ , Pawan K Shahi ₃ , Katherine P Mueller ₄ , Rasa Valiauga ₅ , Kimberly L Edwards ₅ , Cole Bacig ₅ , Stephanie S Steltzer ₄ , Sandhya Srinivasan ₄ , Amr Abdeen ₄ , Evan Cory ₄ , Viswesh Periyasamy ₄ , Alireza Fotuhi Siahpirani ₄ , Edwin M Stone ₆ , Budd A Tucker ₆ , Sushmita Roy ₇ , Bikash R Pattnaik ₈ , Krishanu Saha ₉ , David M Gamm ₁₀

Affiliation

McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA.
McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Department of Pediatrics, University of Wisconsin-Madison, Madison, WI 53792, USA.
Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA.
Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Biostatistics, University of Wisconsin-Madison, Madison, WI 53792, USA.
McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Department of Pediatrics, University of Wisconsin-Madison, Madison, WI 53792, USA; Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI 53705, USA.
McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA; Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI 53705, USA.

Dominantly inherited disorders are not typically considered to be therapeutic candidates for gene augmentation. Here, we utilized induced pluripotent stem cell-derived retinal pigment epithelium (iPSC-RPE) to test the potential of gene augmentation to treat Best disease, a dominant macular dystrophy caused by over 200 missense mutations in BEST1. Gene augmentation in iPSC-RPE fully restored BEST1 calcium-activated chloride channel activity and improved rhodopsin degradation in an iPSC-RPE model of recessive bestrophinopathy as well as in two models of dominant Best disease caused by different mutations in regions encoding ion-binding domains. A third dominant Best disease iPSC-RPE model did not respond to gene augmentation, but showed normalization of BEST1 channel activity following CRISPR-Cas9 editing of the mutant allele. We then subjected all three dominant Best disease iPSC-RPE models to gene editing, which produced premature stop codons specifically within the mutant BEST1 alleles. Single-cell profiling demonstrated no adverse perturbation of retinal pigment epithelium (RPE) transcriptional programs in any model, although off-target analysis detected a silent genomic alteration in one model. These results suggest that gene augmentation is a viable first-line approach for some individuals with dominant Best disease and that non-responders are candidates for alternate approaches such as gene editing. However, testing gene editing strategies for on-target efficiency and off-target events using personalized iPSC-RPE model systems is warranted. In summary, personalized iPSC-RPE models can be used to select among a growing list of gene therapy options to maximize safety and efficacy while minimizing time and cost. Similar scenarios likely exist for other genotypically diverse channelopathies, expanding the therapeutic landscape for affected individuals.

中文翻译：

人类 iPSC 建模揭示了基因型多样化显性黄斑病变中基因治疗的突变特异性反应。

显性遗传疾病通常不被认为是基因增强的治疗候选者。在这里，我们利用诱导多能干细胞衍生的视网膜色素上皮 (iPSC-RPE) 来测试基因增强治疗 Best 疾病的潜力，这是一种由BEST1中 200 多个错义突变引起的显性黄斑营养不良. iPSC-RPE 中的基因增强在隐性 bestrophinopathy 的 iPSC-RPE 模型以及由编码离子结合结构域的不同突变引起的两种显性 Best 疾病模型中完全恢复了 BEST1 钙激活的氯离子通道活性并改善了视紫红质降解。第三种主要的 Best 疾病 iPSC-RPE 模型对基因增强没有反应，但在 CRISPR-Cas9 编辑突变等位基因后显示 BEST1 通道活性正常化。然后，我们对所有三种主要的最佳疾病 iPSC-RPE 模型进行基因编辑，这会在突变体BEST1中产生过早终止密码子等位基因。尽管脱靶分析在一个模型中检测到沉默的基因组改变，但单细胞分析表明在任何模型中都没有对视网膜色素上皮 (RPE) 转录程序造成不利干扰。这些结果表明，对于某些患有显性 Best 疾病的个体来说，基因增强是一种可行的一线方法，而无反应者是基因编辑等替代方法的候选者。然而，有必要使用个性化的 iPSC-RPE 模型系统测试基因编辑策略的目标效率和脱靶事件。总之，个性化 iPSC-RPE 模型可用于在越来越多的基因治疗选项列表中进行选择，以最大限度地提高安全性和有效性，同时最大限度地减少时间和成本。其他基因型不同的通道病可能存在类似的情况，

更新日期：2020-08-06

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11