Dry eye disease in mice activates adaptive corneal epithelial regeneration distinct from constitutive renewal in homeostasis

Abstract

Many epithelial compartments undergo constitutive renewal in homeostasis but activate unique regenerative responses following injury. The clear corneal epithelium is crucial for vision and is renewed from limbal stem cells (LSCs). Using single-cell RNA sequencing, we profiled the mouse corneal epithelium in homeostasis, aging, diabetes, and dry eye disease (DED), where tear deficiency predisposes the cornea to recurrent injury. In homeostasis, we capture the transcriptional states that accomplish continuous tissue turnover. We leverage our dataset to identify candidate genes and gene networks that characterize key stages across homeostatic renewal, including markers for LSCs. In aging and diabetes, there were only mild changes with <15 dysregulated genes. The constitutive cell types that accomplish homeostatic renewal were conserved in DED but were associated with activation of cell states that comprise “adaptive regeneration.” We provide global markers that distinguish cell types in homeostatic renewal vs. adaptive regeneration and markers that specifically define DED-elicited proliferating and differentiating cell types. We validate that expression of SPARC, a marker of adaptive regeneration, is also induced in corneal epithelial wound healing and accelerates wound closure in a corneal epithelial cell scratch assay. Finally, we propose a classification system for LSC markers based on their expression fidelity in homeostasis and disease. This transcriptional dissection uncovers the dramatically altered transcriptional landscape of the corneal epithelium in DED, providing a framework and atlas for future study of these ocular surface stem cells in health and disease.

Multiomic Screening Unravels the Immunometabolic Signatures and Drug Targets of Age-Related Macular Degeneration

“…Interestingly, we also found a significant causal relationship with AMD in our immunome MR analysis. Another clinical study found a significant association between CD16+ monocytes and refractory wet AMD (Lin et al, 2024). Coupled with scRNA-seq results, we hypothesize that the androgen -IL10RA -CD16+monocytes pathway is likely to be a critical immunometabolic pathway in wet AMD.…”

“…Our prior analysis unveiled distinctive immunometabolic factors across various stages and types of AMD, prompting a systematic investigation into immune-related changes. We delved into the peripheral immune milieu at single-cell resolution in 43 AMD patients, encompassing 8 with early AMD, 6 with GA, 16 with CNV, 13 with mixed GA and CNV, alongside 22 healthy controls (Lin et al, 2024). Predominant cell populations were delineated utilizing scRNA-seq data (Figure 3A, S5A).…”

“…The scRNA-seq datasets, encompassing blood samples from both healthy individuals and patients diagnosed with AMD, are made available to the public domain through the Gene Expression Omnibus (GEO) repository, through the accession code GSE222647 (Lin et al, 2024). The initial phase of quality control was executed by applying a set of criteria for cell selection: specifically, only those cells were considered that exhibited a gene expression count ranging from more than 400 to less than 8,000 and an UMI (Unique Molecular Identifier) count that fell between 600 and 120,000.…”

Unlocking the therapeutic potential of the corneal endothelium: Intravital imaging reveals endogenous regenerative capabilities

“…Optimal clustering of cells was performed using the assistance of the clustree R package [47]. Cell type assignment was performed using marker genes defined in murine single cell literature [48][49][50][51]. Volcano plots were created using the R package EnhancedVolcano using differentially expressed gene lists generated by Seurat's FindMarker function (Wilcoxon Rank Sum tests with P < 0.05) [52].…”

Study on the Promotion of hADSCs Migration and Chemotaxis by SDF-1

“…Because of the convenience of obtaining materials, the richness of sources, and the low immunogenicity,3 stem cells have been applied in various fields, such as tissue engineering and regenerative medicine, in recent years 4,5. It has also contributed to the alternative treatment of some ocular diseases 4–9. Many ocular diseases, such as corneal degeneration, retinitis pigmentosa, and diabetic retinopathy, require cell tissue transplantation or vascular reconstruction.…”