Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations
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 fro...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.939774/full |
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| author | Ting Su Liying Liang Lan Zhang Jianing Wang Luyin Chen Caiying Su Jixing Cao Quan Yu Shuai Deng Shuai Deng Hon Fai Chan Hon Fai Chan Shibo Tang Yonglong Guo Yonglong Guo Jiansu Chen Jiansu Chen Jiansu Chen |
| author_facet | Ting Su Liying Liang Lan Zhang Jianing Wang Luyin Chen Caiying Su Jixing Cao Quan Yu Shuai Deng Shuai Deng Hon Fai Chan Hon Fai Chan Shibo Tang Yonglong Guo Yonglong Guo Jiansu Chen Jiansu Chen Jiansu Chen |
| author_sort | Ting Su |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:57:05Z |
| format | Article |
| id | doaj.art-48118f4b207649a0aa8ef21185b5bed1 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-11T09:57:05Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-48118f4b207649a0aa8ef21185b5bed12022-12-22T04:30:32ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-09-011010.3389/fbioe.2022.939774939774Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutationsTing Su0Liying Liang1Lan Zhang2Jianing Wang3Luyin Chen4Caiying Su5Jixing Cao6Quan Yu7Shuai Deng8Shuai Deng9Hon Fai Chan10Hon Fai Chan11Shibo Tang12Yonglong Guo13Yonglong Guo14Jiansu Chen15Jiansu Chen16Jiansu Chen17Department of Ophthalmology, First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, ChinaDepartment of Ophthalmology, First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, ChinaDepartment of Ophthalmology, First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, ChinaKey Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, ChinaKey Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, ChinaDepartment of Ophthalmology, First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, ChinaDepartment of Ophthalmology, First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, ChinaCentric Laboratory, Medical College, Jinan University, Guangzhou, ChinaInstitute for Tissue Engineering and Regenerative Medicine, Chinese University of Hong Kong, Hong Kong, ChinaKey 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, ChinaInstitute for Tissue Engineering and Regenerative Medicine, Chinese University of Hong Kong, Hong Kong, ChinaKey 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, ChinaAier Eye Institute, Changsha, ChinaDepartment of Ophthalmology, First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou, ChinaInstitute of Ophthalmology, Medical College, Jinan University, Guangzhou, ChinaKey Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, ChinaAier Eye Institute, Changsha, ChinaRetinitis 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.https://www.frontiersin.org/articles/10.3389/fbioe.2022.939774/fullretinitis pigmentosaUSH2Ainduced pluripoten stem cellsretinal organoidsmicrofludic chip |
| spellingShingle | Ting Su Liying Liang Lan Zhang Jianing Wang Luyin Chen Caiying Su Jixing Cao Quan Yu Shuai Deng Shuai Deng Hon Fai Chan Hon Fai Chan Shibo Tang Yonglong Guo Yonglong Guo Jiansu Chen Jiansu Chen Jiansu Chen Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations Frontiers in Bioengineering and Biotechnology retinitis pigmentosa USH2A induced pluripoten stem cells retinal organoids microfludic chip |
| title | Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations |
| title_full | Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations |
| title_fullStr | Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations |
| title_full_unstemmed | Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations |
| title_short | Retinal organoids and microfluidic chip-based approaches to explore the retinitis pigmentosa with USH2A mutations |
| title_sort | retinal organoids and microfluidic chip based approaches to explore the retinitis pigmentosa with ush2a mutations |
| topic | retinitis pigmentosa USH2A induced pluripoten stem cells retinal organoids microfludic chip |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.939774/full |
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