Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations.
Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations.
复制标题
视网膜类器官和基于微流控芯片的方法探索 USH2A 突变引起的视网膜色素变性
DOI：
10.3389/fbioe.2022.939774
复制
发表时间：
2022
影响因子：
5.7
通讯作者：
Chen, Jiansu
中科院分区：
文献类型：
--
作者：
Su, Ting;Liang, Liying;Zhang, Lan;Wang, Jianing;Chen, Luyin;Su, Caiying;Cao, Jixing;Yu, Quan;Deng, Shuai;Chan, Hon Fai;Tang, Shibo;Guo, Yonglong;Chen, Jiansu
视网膜色素变性（RP）是全球范围内视力受损和失明的主要原因，医疗选择有限。USH 2A突变是非综合征型RP的最常见原因之一。在这项研究中，我们从RP患者的诱导多能干细胞（iPSCs）中开发了视网膜类器官（RO）和视网膜色素上皮（RPE）细胞，以建立可持续的体外RP疾病模型。RT-qPCR、蛋白质印迹和免疫荧光染色评估显示USH 2A突变诱导iPSC和RO的凋亡以及细胞外基质（ECM）组分的缺乏。转录组学和蛋白质组学研究结果表明，异常ECM-受体相互作用可通过PI 3 K-Akt途径导致USH 2A突变的RO凋亡。为了优化视网膜色素上皮细胞的培养条件，我们制作了一个微流控芯片，用于视网膜色素上皮细胞与视网膜色素上皮细胞的共培养。结果表明，该灌注系统能有效提高RO的存活率。此外，ECM成分，如层粘连蛋白和IV型胶原蛋白的RO在RP组上调相比，那些保持在静态培养。这些发现说明了微流控芯片结合RO技术在RP疾病建模中的潜力。
Retinitis pigmentosa (RP) is a leading cause of vision impairment and blindness worldwide, with limited medical treatment options. USH2A mutations are one of the most common causes of non-syndromic RP. In this study, we developed retinal organoids (ROs) and retinal pigment epithelium (RPE) cells from induced pluripotent stem cells (iPSCs) of RP patient to establish a sustainable in vitro RP disease model. RT-qPCR, western blot, and immunofluorescent staining assessments showed that USH2A mutations induced apoptosis of iPSCs and ROs, and deficiency of the extracellular matrix (ECM) components. Transcriptomics and proteomics findings suggested that abnormal ECM-receptor interactions could result in apoptosis of ROs with USH2A mutations via the PI3K-Akt pathway. To optimize the culture conditions of ROs, we fabricated a microfluidic chip to co-culture the ROs with RPE cells. Our results showed that this perfusion system could efficiently improve the survival rate of ROs. Further, ECM components such as laminin and collagen IV of ROs in the RP group were upregulated compared with those maintained in static culture. These findings illustrate the potential of microfluidic chip combined with ROs technology in disease modelling for RP.
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DOI：
10.1038/s41578-021-00279-y
发表时间：
2021
期刊：
Nature reviews. Materials
影响因子：
--
作者：
Hofer M;Lutolf MP
通讯作者：
Lutolf MP
影响因子：
14
作者：
Dorgau, Birthe;Felemban, Majed;Lako, Majlinda
通讯作者：
Lako, Majlinda
影响因子：
4.4
作者：
Lopez-Luppo, Mariana;Nacher, Victor;Ruberte, Jesus
通讯作者：
Ruberte, Jesus
影响因子：
5.9
作者：
Huang, Kang-Chieh;Wang, Mong-Lien;Chiou, Shih-Hwa
通讯作者：
Chiou, Shih-Hwa
影响因子：
2.9
作者：
Battacharya, G;Cosgrove, D
通讯作者：
Cosgrove, D