Cell atlas of the human ocular anterior segment: Tissue-specific and shared cell types

Proceedings of the National Academy of Sciences, Год журнала: 2022, Номер 119(29)

Опубликована: Июль 15, 2022

The anterior segment of the eye consists cornea, iris, ciliary body, crystalline lens, and aqueous humor outflow pathways. Together, these tissues are essential for proper functioning eye. Disorders vision have been ascribed to defects in all them; some disorders, including glaucoma cataract, among most prevalent causes blindness world. To characterize cell types that compose tissues, we generated an atlas human using high-throughput single-nucleus RNA sequencing (snRNAseq). We profiled 195,248 nuclei from nondiseased six donors, identifying >60 types. Many were discrete, whereas others, especially lens formed continua corresponding known developmental transitions persist adulthood. Having each tissue separately, performed integrated analysis entire segment, revealing unique a single structure, others shared across tissues. was then used investigate type-specific expression patterns more than 900 ocular disease genes identified through either Mendelian inheritance or genome-wide association studies.

Язык: Английский

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Mass Spectrometric Detection of Keratins in Tear Fluid

Experimental Eye Research, Год журнала: 2025, Номер 251, С. 110231 - 110231

Опубликована: Янв. 4, 2025

Язык: Английский

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Human conjunctiva organoids to study ocular surface homeostasis and disease

Cell stem cell, Год журнала: 2024, Номер 31(2), С. 227 - 243.e12

Опубликована: Янв. 11, 2024

The conjunctival epithelium covering the eye contains two main cell types: mucus-producing goblet cells and water-secreting keratinocytes, which present mucins on their apical surface. Here, we describe long-term expanding organoids air-liquid interface representing mouse human conjunctiva. A single-cell RNA expression atlas of primary cultured conjunctiva reveals that keratinocytes express multiple antimicrobial peptides identifies tuft cells. IL-4/-13 exposure increases differentiation drastically modifies secretome. Human NGFR+ basal are identified as bipotent stem Conjunctival cultures can be infected by herpes simplex virus 1 (HSV1), adenovirus 8 (hAdV8), SARS-CoV-2. HSV1 infection was reversed acyclovir addition, whereas hAdV8 infection, lacks an approved drug therapy, inhibited cidofovir. We document transcriptional programs induced hAdV8. Finally, transplanted. Together, organoid enable study (patho)-physiology.

Язык: Английский

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Cell therapy in the cornea: The emerging role of microenvironment

Progress in Retinal and Eye Research, Год журнала: 2024, Номер 102, С. 101275 - 101275

Опубликована: Май 24, 2024

Язык: Английский

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Single-cell RNA sequencing reveals the complex cellular niche of pterygium

The Ocular Surface, Год журнала: 2024, Номер 32, С. 91 - 103

Опубликована: Янв. 28, 2024

Язык: Английский

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RNA-Seq–based transcriptome analysis of corneal endothelial cells derived from patients with Fuchs endothelial corneal dystrophy

Scientific Reports, Год журнала: 2023, Номер 13(1)

Опубликована: Май 27, 2023

Fuchs endothelial corneal dystrophy (FECD) is the most common inherited disease. Fibrillar focal excrescences called guttae and edema due to cell death result in progressive vision loss. Multiple genetic variants have been reported, but pathogenesis of FECD not fully understood. In this study, we used RNA-Seq analyze differential gene expression endothelium obtained from patients with FECD. Differential analysis transcriptomic profiles revealed that 2366 genes (1092 upregulated 1274 downregulated genes) was significantly altered compared healthy subjects. Gene ontology demonstrated an enrichment involved extracellular matrix (ECM) organization, response oxidative stress, apoptotic signaling. Several pathway analyses consistently indicated dysregulation ECM-associated pathways. Our findings support previously proposed underlying mechanisms, including stress apoptosis cells, as well phenotypic clinical hallmark ECM deposits. Further investigation focusing on differentially expressed related these pathways might be beneficial for elucidating mechanisms developing novel therapies.

Язык: Английский

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Therapeutic Strategies for Restoring Perturbed Corneal Epithelial Homeostasis in Limbal Stem Cell Deficiency: Current Trends and Future Directions

Cells, Год журнала: 2022, Номер 11(20), С. 3247 - 3247

Опубликована: Окт. 16, 2022

Limbal stem cells constitute an important cell population required for regeneration of the corneal epithelium. If insults to limbal or their niche are sufficiently severe, a disease known as deficiency occurs. In absence functioning cells, vision-compromising conjunctivalization epithelium occurs, leading opacification, inflammation, neovascularization, and chronic scarring. transplantation is standard treatment unilateral cases deficiency, but bilateral require use cultured non-limbal autologous allogeneic transplantation. Herein we review current therapeutic utilization cells. We also describe several markers that impact phenotype function discuss possibility modulating other sources facilitate development novel interventions. finally consider hurdles widespread adoption these proposed methodologies how they can be overcome realize vision-restoring

Язык: Английский

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Single-Cell Transcriptomics Reveals Cellular Heterogeneity and Complex Cell–Cell Communication Networks in the Mouse Cornea

Investigative Ophthalmology & Visual Science, Год журнала: 2023, Номер 64(13), С. 5 - 5

Опубликована: Окт. 4, 2023

To generate a single-cell RNA-sequencing (scRNA-seq) map and construct cell-cell communication networks of mouse corneas.C57BL/6 corneas were dissociated to single cells subjected scRNA-seq. Cell populations clustered annotated for bioinformatic analysis using the R package "Seurat." Differential expression patterns validated spatially mapped with whole-mount immunofluorescence staining. Global intercellular signaling constructed CellChat.Unbiased clustering scRNA-seq transcriptomes 14,732 from 40 revealed 17 cell clusters six major types: nine epithelial cell, three keratocyte, two corneal endothelial one each immune vascular fibroblast clusters. The subtypes included quiescent limbal stem cells, transit-amplifying differentiated minor conjunctival CellChat provided an atlas complex communications among all types.We complete transcriptomic cross-talk types cornea, which can be used as foundation further research on cornea. This study also deepens understanding cellular heterogeneity heterotypic interaction within corneas.

Язык: Английский

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Sox9 marks limbal stem cells and is required for asymmetric cell fate switch in the corneal epithelium

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Апрель 11, 2024

Abstract Adult tissues with high cellular turnover require a balance between stem cell renewal and differentiation, yet the mechanisms underlying this equilibrium are unclear. The cornea exhibits polarized lateral flow of progenitors from peripheral niche to center; attributed differences in fate. To identify genes that critical for regulating asymmetric fates limbal cells their transient amplified progeny central cornea, we utilized an vivo cycle reporter isolate proliferating basal across anterior ocular surface epithelium performed single-cell transcriptional analysis. This strategy greatly increased resolution revealed distinct identities unique expression profiles structural transcription factors. We focused on Sox9; factor implicated regulation various organs. Sox9 was found be differentially expressed corneal. Lineage tracing analysis confirmed marks long-lived conditional deletion led abnormal differentiation squamous metaplasia cornea. These data suggest requirement switch fate commitment toward as exit niche. By inhibiting terminal corneal forcing them into perpetual symmetric divisions, replicated loss-of-function phenotype. Our findings reveal essential role spatial is required sustain tissue homeostasis.

Язык: Английский

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Single-cell transcriptomic profiling of rat iridocorneal angle at perinatal stages: revisiting the development of periocular mesenchyme

Experimental Eye Research, Год журнала: 2025, Номер unknown, С. 110249 - 110249

Опубликована: Янв. 1, 2025

Язык: Английский

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