Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations

Ting Su; Liying Liang; Lan Zhang; Jianing Wang; Luyin Chen; Caiying Su; Jixing Cao; Quan Yu; Shuai Deng; Hon Fai Chan; Shibo Tang; Yonglong Guo; Jiansu Chen

doi:10.3389/fbioe.2022.939774

Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations

Front Bioeng Biotechnol. 2022 Sep 14:10:939774. doi: 10.3389/fbioe.2022.939774. eCollection 2022.

Authors

Ting Su¹, Liying Liang¹, Lan Zhang¹, Jianing Wang², Luyin Chen², Caiying Su¹, Jixing Cao¹, Quan Yu³, Shuai Deng^{4

5}, Hon Fai Chan^{4

5}, Shibo Tang⁶, Yonglong Guo^{1

7}, Jiansu Chen^{8

2

6}

Affiliations

¹ Department of Ophthalmology, First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, China.
² Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China.
³ Centric Laboratory, Medical College, Jinan University, Guangzhou, China.
⁴ Institute for Tissue Engineering and Regenerative Medicine, Chinese University of Hong Kong, Hong Kong, China.
⁵ Key Laboratory for Regenerative Medicine of the Ministry of Education of China, Ministry of Education of China, School of Biomedical Sciences, Faculty of Medicine, Chinese University of Hong Kong, Hong Kong, China.
⁶ Aier Eye Institute, Changsha, China.
⁷ College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
⁸ Institute of Ophthalmology, Medical College, Jinan University, Guangzhou, China.

Abstract

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.

Keywords: USH2A; induced pluripoten stem cells; microfludic chip; retinal organoids; retinitis pigmentosa.