Eye on a Dish Models to Evaluate Splicing Modulation.
Eye on a Dish Models to Evaluate Splicing Modulation.
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DOI：
10.1007/978-1-0716-2010-6_16
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发表时间：
2022-01-01
期刊：
影响因子：
--
通讯作者：
Cheetham, Michael E
中科院分区：
文献类型：
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作者：
Hau, Kwan-Leong;Lane, Amelia;Cheetham, Michael E
遗传性视网膜营养不良，如Leber先天性黑色素、Stargardt病和视网膜色素变性，以光感受器功能障碍和死亡为特征，目前治疗方案很少。诱导多能干细胞(IPSC)技术和分化方法的最新技术进步意味着现在可以在体外研究人类光感受器。例如，视网膜类有机物为研究人类视网膜的发育和培养皿中疾病的机制提供了一个平台，也是细胞移植的潜在来源。在这里，我们描述了3D培养的分化方案，这种培养产生含有光感受器的视网膜类器官，外节不发育。这些协议可用作了解视网膜疾病机制和测试潜在治疗方法的模型，包括反义寡核苷酸(AON)以改变基因表达或RNA处理。这种“培养皿中的视网膜”模型非常适合与AON一起使用，因为类器官在正确的基因组和细胞环境中概括了患者的突变，以测试潜在的疗效，并检查通往临床的翻译路径上的非靶点影响。
Inherited retinal dystrophies, such as Leber congenital amaurosis, Stargardt disease, and retinitis pigmentosa, are characterized by photoreceptor dysfunction and death and currently have few treatment options. Recent technological advances in induced pluripotent stem cell (iPSC) technology and differentiation methods mean that human photoreceptors can now be studied in vitro. For example, retinal organoids provide a platform to study the development of the human retina and mechanisms of diseases in the dish, as well as being a potential source for cell transplantation. Here, we describe differentiation protocols for 3D cultures that produce retinal organoids containing photoreceptors with rudimentary outer segments. These protocols can be used as a model to understand retinal disease mechanisms and test potential therapies, including antisense oligonucleotides (AONs) to alter gene expression or RNA processing. This "retina in a dish" model is well suited for use with AONs, as the organoids recapitulate patient mutations in the correct genomic and cellular context, to test potential efficacy and examine off-target effects on the translational path to the clinic.