Single-Cell Transcriptomic Analysis Reveals Dynamic Cellular Processes in Corneal Epithelium During Wound Healing in Cynomolgus Monkeys

被引:0
作者
Zhou, Ming [1 ]
Shi, Zhuo-Xing [1 ]
Liu, Zhong [1 ]
Ke, Shu-Rui [1 ]
Wang, Chao-Yang [1 ]
Liang, Xiao-Lin [1 ]
Hu, Qiu-Ling [1 ]
Zhang, Qi-Kai [1 ]
Wang, Dong-Liang [1 ]
Sun, Li [1 ]
Lin, Yu-Heng [1 ]
Dai, Qi [2 ]
Zheng, Ying-Feng [1 ,3 ]
机构
[1] Sun Yat Sen Univ, Zhongshan Ophthalm Ctr, State Key Lab Ophthalmol, Guangdong Prov Key Lab Ophthalmol & Visual Sci, Guangzhou 510060, Peoples R China
[2] Zhejiang Sci Tech Univ, Coll Life Sci & Med, Hangzhou, Peoples R China
[3] Chinese Acad Med Sci, Res Unit Ocular Dev & Regenerat, Beijing 100085, Peoples R China
基金
中国国家自然科学基金;
关键词
corneal wound healing; single-cell transcriptome sequencing; epithelial cells; extracellular matrix (ECM); thrombospondin-1 ( THBS1 ); STEM-CELLS; BASEMENT-MEMBRANE; THROMBOSPONDIN-1; HOMEOSTASIS; EXPRESSION; APOPTOSIS; MODELS; REPAIR;
D O I
10.1167/iovs.65.11.43
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
P URPOSE . Corneal wounding healing is critical for maintaining clear vision, however, a complete understanding of its dynamic regulatory mechanisms remains elusive. Here, we used single-cell RNA sequencing (scRNA-seq) to analyze the cellular activities and transcriptional changes of corneal limbal epithelial cells at different stages after wound healing in cynomolgus monkeys, which exhibit a closer transcriptomic similarity to humans. M ETHODS . Corneal limbal tissues were collected during uninjured, 1-day and 3-day healing stages, dissociated into single cells, and subjected to scRNA-seq using the 10x x Genomics platform. Cell types were clustered by graph-based visualization methods and unbiased computational analysis. Additionally, cell migration assays and immunofluorescent staining were performed on cultured human corneal epithelial cells. R ESULTS . We characterized nine cell clusters by scRNA-seq analysis of the cynomolgus monkey corneal epithelium. By comparing heterogeneous transcriptional changes in major cell types during corneal healing, we highlighted the importance of limbal epithelial cells (LEPCs) and basal epithelial cells (BEPCs) in extracellular matrix (ECM) formation and wound healing, as well as suprabasal epithelial cells (SEPCs) in epithelial differentiation during the healing processes. We further identified five different sub-clusters in LEPC, including the transit amplifying cell (TAC) sub-cluster that promotes early healing through the activation of thrombospondin-1 ( THBS1 ) expression. C ONCLUSIONS . Our study represents the first comprehensive exploration of the detailed transcriptome profile of individual corneal cells during the wound healing process in nonhuman primates. We demonstrate the intricate mechanisms involved in corneal healing and provide a promising avenue for potential therapies in corneal wound healing.
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页数:17
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