MiR-141-3p regulates the expression of corneal epithelial differentiation
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ABSTRACT:
ORGANISM(S): Oryctolagus cuniculus
PROVIDER: GSE264014 | GEO | 2024/04/20
REPOSITORIES: GEO
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Project description:Corneal epithelial RCE1(5T5) cells follow a sequential process that leads to the formation of a 4-5 layered stratified epithelium with a gene expression pattern similar to that shown in primary cultures of corneal epithelial cells. We have identified three different developmental stages during the differentiation of the rabbit corneal epithelial cell line RCE1(5T5), and thus, analyzed the transcriptome of such different stages in order to understand the regulation of the corneal epithelial cell differentiation. This analysis has led to identify the global expression of gene signatures associated to such differentiation stages.
2020-12-29 | GSE147779 | GEO
Project description:miR-141-3p regulates the expression of corneal epithelial differentiation
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2010-12-11 | E-GEOD-26009 | biostudies-arrayexpress
Project description:The corneal epithelium is a stratified squamous epithelium that protects the eye from mechanical and toxic stress. In this system, cells are continually being sloughed off from the surface of the eye, and replenished by proliferation from corneal stem cells in the limbus, which migrate in and differentiate. During tissue development and homeostasis, distal regulatory regions called enhancers, which contain binding sites for numerous transcription factors, help to control and coordinate gene expression in a temporal and spatial specific manner. To identify enhancers in cornea epithelia, we performed ChIP-Seq with antibodies to the histone marks H3K4me3, H3K4me1, and H3K27ac in primary human corneal epithelial cells. Active enhancers were defined as having high levels of H3K4me1 and H3K27ac, and low levels of H3K4me3: 12,900 such regions were identified in the corneal epithelial cells. In combination with primary cell histone modification ChIP-Seq data from BROAD, 2946 active enhancer regions were identified that are unique to the corneal epithelial cells: 1551 of theses 2946 also overlie regions of high evolutionary constraint. Motif enrichment analysis of these regions has revealed binding sites for a number of transcription factors, highlighting their roles as key regulators of corneal epithelial development.
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Project description:The regenerative capacity of corneal endothelial cells (CECs) differs between species; in bigger mammals such as humans and sheep, CECs are arrested in a non-proliferative state. Damage to these cells can compromise its function leading to corneal opacity and impaired vision. Corneal transplantation is the current treatment for the recovery of a clear eyesight, but the donor tissue demand is higher than the offer and there is the need to develop novel treatment modalities. In contrast, rabbit CECs retain a high proliferative profile and have the capacity to repopulate the endothelium upon injury. There is currently a lack of fundamental knowledge to explain the difference in the proliferation capacity of these cells across species. Gaining information on the transcriptomic differences across species could allow the identification of crucial proliferation drivers of CEC proliferation to develop novel regenerative medicine therapies. In this study we have analyzed at the transcriptomic level corneal endothelial samples originating from human, sheep, and rabbit. To understand the differences across the CECs proliferation capacity in each species, we have generated an automated pipeline for the analysis of pathways with different activity. Our results revealed that 52 pathways had commonly different activity when comparing species with non-proliferative endothelium (human and sheep) to species with proliferative endothelium (rabbit).
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Project description:MicroRNAs, small endogenous non-coding RNAs, are involved in the regulation of epidermal homeostasis. Among them, miR-203 was the most described and expressed in human epidermis, promoting keratinocyte differentiation by repressing genes involved in proliferation. To identify other miRNAs involved in this process, the miRNomes of normal human keratinocytes cultured in monolayer (2D) or in 3D reconstructed skin were compared. Besides miR-203, mR-141 was one of the most expressed miRNAs in 3D culture and was overexpressed in 3D vs 2D condition, i.e. during keratinocyte differentiation. Functional experiments revealed that, mostly expressed in the basal layer, miR-141 decreased keratinocyte proliferation and clonogenicity while promoting their differentiation. Target prediction algorithm coupled with transcriptomic data of keratinocytes overexpressing miR-141, as well as 3’UTR luciferase assays enabled to evidence CCND2 mRNA as a direct target of miR-141, leading to its down regulation by miR-141 overexpression. Finally, CCND2 silencing decreased keratinocyte proliferation and induced their differentiation, revealing that miR-141 action was mediated by CCND2. MiR-141 features were also compared with miR-203 in parallel experiments. Although miR-141 displayed similar functions to the ones of miR-203, it exhibited different localization and targets, suggesting a joint participation of miR-141 and miR-203 to engage and maintain keratinocyte towards differentiation, respectively.
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Project description:We successfully induced corneal epithelial cells from human iPSCs. Then, we perfomed global expression analysis using microarray to compare the character of hiPSC-derived corneal epithelial cells with that of the other kinds of cells. Total RNA was obtained from human iPSCs (hiPSCs), human iPSC-derived corneal epithelial cells (hiCECs), human corneal limbal epithelial cells (HCECs), human oral keratinocytes (HOKs), human dermal fibroblasts (HDFs) and six weeks-differentiated hiPSCs (hiPSC-derived ocular surface ectoderm, OSE) using the QIAZol reagent. A microarray analysis using Sure Print G3 human 8x60K slides (Agilent technologies) was performed at Takara Bio (Shiga, Japan).
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Project description:Mycotic leratitis is the corneal inflammation predominantly caused by Fusarium and Aspergillus species. Corneal epithelium is the earliest cell type encounter the invading pathogen. The innate immune responses of human corneal epithelial cells against Aspergillus flavus is not known. Here we studied the role of human corneal epithelial cells against Aspergillus flavus infection. The results showed that corneal epithelial cells internalized Aspergillus flavus conidia through actin mediated polymerization surrounding the conidia. Further the actin inhibitor cytochalasin D treatment reduced the formation actin ring around the conidia. The engulfed conidia acquired endosomal proteins as revealed by immunofluorescence analsyisis. Mass spectromtery of phagosomal proteins confirmed the recruitment of endosomal proteins and other proteins involved in phagocytosis. These results show the involvement of corneal epithelial cells in anti fungal defense.
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