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Application of single-cell RNA sequencing in diagnosis and treatment of corneal and retinal diseases
Huang Chang, Ying Peixi, Yi Guoguo, Fu Sheng, Wang Yan, Guo Xi, Zhang Yuxi, Fu Min
2022, 26 (13):
2106-2113.
doi: 10.12307/2022.341
BACKGROUND: Single-cell RNA sequencing, as a new generation of mRNA sequencing method, has gradually matured and been applied in ophthalmic research after more than ten years of development.
OBJECTIVE: To review the development of single cell RNA sequencing and its application and progress in ophthalmology.
METHODS: The relevant articles in PubMed, Sciencedirect, and Web of Science databases were searched by computer. English key words were “single-cell RNA-seq, retina, cornea, gene marker”. The retrieval time was from January 1, 2006 to January 31, 2021. The articles were screened initially through reading and analysis to exclude duplicates and low-relevance articles. Finally, 80 articles were included for results analysis.
RESULTS AND CONCLUSION: (1) Single cell RNA sequencing is on the single cell level, of genome, transcriptome, and epigenome high-throughput sequencing analysis technology. Compared with the conventional sequencing method, it can be further applied to study the differences between the cells, explain cell subtype and specific expression genes, to explore the disease occurrence, development and resistance mechanism, and provide new targets for the immune therapy. (2) Currently, there are many branches of single-cell RNA sequencing, such as STRT-seq, Smart-seq, CEL-seq, MARS-seq, Cyto-seq, Drop-seq, inDrop-seq, and Seq-Well. These technologies have their own advantages and application fields. For example, STRT-seq is only at the 5’ end and does not have chain specificity, whereas Samrt-seq transcriptional coverage is full-length. (3) In the field of ophthalmology, single-cell RNA sequencing is mainly used in the study of cornea and retina. There are new scientific discoveries in the identification of new subtypes of eye cells and the diagnosis of ophthalmic diseases and potential immunotherapy. A few are used in the study of lens diseases and uveal melanoma. (4) The research of single-cell RNA sequencing technology in the field of cornea is mainly about keratoconus disease and corneal epithelial cell gene expression. Notch1 and PLLP genes were proven to be related to the pathogenesis of keratoconus. The expression of Krt15, Fzd7, and Krt17 genes increased in corneal basal cells, while Sox9, Actn1, and Anxa3 were mainly expressed in basal epithelial progenitor cells. In addition, retinal tissue is a kind of tissue that is easy to obtain and observe. In recent years, single-cell RNA sequencing technology has been applied to study the gene expression of normal retinal tissue cell subtypes and retinal-related diseases, such as age-related macular degeneration and retinoblastoma. (5) Single-cell RNA sequencing technology can accurately reflect the gene expression of a single cell. In recent years, it has been used to study the gene expression, cell subtypes and signaling pathways of cornea and retina, and will play an important role in the treatment of ophthalmic diseases in the future.
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