Targeted proteomic and bioinformatic investigation of extracellular matrix remodeling in hAEC-EV-mediated corneal repair
Ontology highlight
ABSTRACT:
ORGANISM(S): Oryctolagus Cuniculus
SUBMITTER: 
Hong Liu
PROVIDER: PXD075526 | iProX | Wed Mar 11 00:00:00 GMT 2026
REPOSITORIES: iProX
Publications
Targeted proteomic and bioinformatic investigation of extracellular matrix remodeling in hAEC-EV-mediated corneal repair.
Hu Shuqin S Qiu Ting T Liu Hong H
Frontiers in bioengineering and biotechnology 20260318
<h4>Introduction</h4>Human amniotic epithelial cell-derived extracellular vesicles (hAEC-EVs) have shown therapeutic potential in corneal injury repair; however, the underlying molecular mechanisms, particularly those related to extracellular matrix (ECM) remodeling, remain incompletely understood.<h4>Methods</h4>A proteomic and bioinformatic strategy was applied to analyze ECM-related molecular alterations in corneal tissues following alkali injury and hAEC-EV treatment. Differentially expresse ...[more]
Similar Datasets
Project description:Transplantation of amniotic membrane-expanded limbal epithelium (AMLE) in place of donor tissue grafts results in significantly improved outcomes for patients suffering from severe limbal stem cell deficiency; however the reasons for such superior results are unclear. The purpose of this study was to identify transcriptional gene profiles specific to AMLE and donor central corneal epithelium (CE), which may contribute to the divergent clinical outcomes observed following transplant. Limbal fibroblasts which underlie the epithelium and secrete extracellular matrix proteins following injury/surgery were also profiled. Using cell culture, immunofluorescence, microarray gene expression profiling and qRT-PCR validation; this study aims to identify enriched biological processes and pathways which characterise AMLE and CE tissues. We hope the study outcomes will shed light onto the factors which contribute to provide the improved clinical outcomes associated with AMLE transplantation. Gene expression profiling of three central corneal buttons, three amniotic membrane-expanded limbal epithelial cultures and three limbal fibroblast cultures
2014-10-02 | E-GEOD-56421 | biostudies-arrayexpress
Project description:Dr. Panjwani's laboratory is focusing on the mechanism by which galectins-3 and 7 mediate corneal epithelial cell migration. We are currently performing studies to: (i) identify and characterize the corneal epithelial cell surface and extracellular matrix (ECM) molecules which serve as counterreceptors of galectin-3 and -7, to establish whether the lectins modulate corneal epithelial cell migration by binding to well known integrins, growth factor receptors, and/or ECM molecules and (ii) determine whether galectin-3 mediates corneal epithelial cell migration indirectly by modulating the expression of key adhesion and/or signal transduction molecules by using small interfering RNA, cDNA microarrays and glycogene arrays. We have prepared three independent preparations of total RNA of corneal epithelial cells from WT mice (total six samples) for analysis of glycogene expression. Samples are Normal Cornea (Left eye) and Laser ablation + 16-18 hours healing (right eye)
2010-12-11 | E-GEOD-26009 | biostudies-arrayexpress
Project description:The human cornea relies on its extracellular matrix (ECM) properties, with corneal stromal fibroblasts (CSFs) crucial for ECM remodeling. We studied the effects of nanofibrillar materials on CSFs using a decellularized human amniotic membrane (dHAM) and fibrillar collagen scaffold (FCS). FCS enhances focal adhesions (FAs), activating mechanotransduction signals through FAK autophosphorylation and YAP1 nuclear translocation. Nanofibrillar topography induces gene expression changes related to tissue remodeling and collagen production. Here, we performed shotgun proteomics analysis (DDA-PASEF in TimsToF Pro) to analyze the impact of FCS topography on CSFs protein expression.
2024-09-02 | PXD048096 | Pride
Project description:Corneal injuries remain a major cause of consultation in the ophthalmology clinics worldwide. Repair of corneal wounds is a complex mechanism that involves cell death, migration, proliferation, differentiation, and extracellular matrix (ECM) remodeling. In the present study, we used a tissue-engineered, two-layers (epithelium and stroma) human cornea as a biomaterial to study both the cellular and molecular mechanisms of wound healing. Gene profiling on microarrays revealed important alterations in the pattern of genes expressed by tissue-engineered corneas in response to wound healing. Expression of many MMPs-encoding genes was shown by microarray and qPCR analyses to increase in the migrating epithelium of wounded corneas. Many of these enzymes were converted into their enzymatically active form as wound closure proceeded. In addition, expression of MMPs by human corneal epithelial cells (HCECs) was affected both by the stromal fibroblasts and the collagen-enriched ECM they produce. Most of all, results from mass spectrometry analyses provided evidence that a fully stratified epithelium is required for proper synthesis and organization of the ECM on which the epithelial cells adhere. In conclusion, and because of the many characteristics it shares with the native cornea, this human two layers corneal substitute may prove particularly useful to decipher the mechanistic details of corneal wound healing. Primary cultures of human corneal epithelial cells cultivated on BSA (number of replicates: 7), Collagen type I (number of replicates: 2), Collagen type IV (number of replicates: 2), Fibronectin (number of replicates: 2), Tenascin C (number of replicates: 2) and Laminin (number of replicates: 2) matrix. Central, internal and external ring of wounded Tissue-engineered human cornea.
2015-11-25 | E-GEOD-75336 | biostudies-arrayexpress
Project description:Transplantation of amniotic membrane-expanded limbal epithelium (AMLE) in place of donor tissue grafts results in significantly improved outcomes for patients suffering from severe limbal stem cell deficiency; however the reasons for such superior results are unclear. The purpose of this study was to identify transcriptional gene profiles specific to AMLE and donor central corneal epithelium (CE), which may contribute to the divergent clinical outcomes observed following transplant. Limbal fibroblasts which underlie the epithelium and secrete extracellular matrix proteins following injury/surgery were also profiled. Using cell culture, immunofluorescence, microarray gene expression profiling and qRT-PCR validation; this study aims to identify enriched biological processes and pathways which characterise AMLE and CE tissues. We hope the study outcomes will shed light onto the factors which contribute to provide the improved clinical outcomes associated with AMLE transplantation.
2014-10-02 | GSE56421 | GEO
Project description:The corneal epithelial barrier maintains the metabolic activities of the ocular surface by regulating membrane transporters and metabolic enzymes responsible for the homeostasis of the eye as well as the pharmacokinetic behavior of drugs. Despite its importance, no established biomimetic in vitro methods are available to perform the spatiotemporal investigation of metabolism and determine the transportation of endogenous and exogenous molecules across the corneal epithelium barrier. This study introduces multiple corneal epitheliums on a chip namely, Corneal Epithelium on a Chip (CEpOC), which enables the spatiotemporal collection as well as analysis of micro-scaled extracellular metabolites from both the apical and basolateral sides of the barriers. Longitudinal samples collected during 48 h period were analyzed using untargeted liquid chromatography-mass spectrometry metabolomics method, and 104 metabolites were annotated. We observed the spatiotemporal secretion of biologically relevant metabolites (i.e., antioxidant, glutathione and uric acid) as well as the depletion of essential nutrients such as amino acids and vitamins mimicking the in vivo molecules trafficking across the human corneal epithelium. Through the shifts of extracellular metabolites and quantitative analysis of mRNA associated with transporters, we were able to investigate the secretion and transportation activities across the polarized barrier in a correlation with the expression of corneal transporters. Thus, CEpOC can provide a non-invasive, simple, yet effectively informative method to determine pharmacokinetics and pharmacodynamics as well as to discover novel biomarkers for drug toxicological and safety tests as advanced experimental model of the human corneal epithelium.
2021-07-13 | MTBLS2274 | MetaboLights
Project description:Fuchs’ endothelial corneal dystrophy is major corneal disorder in the western world affecting the innermost part of the cornea, which leads to visual impairment. The morphological changes observed in Fuchs’ endothelial corneal dystrophy is well described, however, much less in known of the pathology at the molecular level. As the morphological changes observed in the cornea is profound in the extracellular matrix we sought to determine in protein profiles and changes herein in the Descement’s membrane and endothelium layer of Fuchs’ endothelial conrneal dystrophy patients when compared to healthy control tissue. Using the extracted ion chromatogram label-free MS based quantification method we quantified approximately the 50 most abundant proteins of the Descemet’s membrane and endothelial layer in in patient and control tissue. In addition, using the isobaric tag for relative and absolute quantification MS method resulted in a total of 22 regulated proteins of which the majority were extracellular proteins known to be involved in proper assembly and modulation of the basement membrane in other tissues. Many of the regulated proteins were furthermore among the most abundant proteins quantified. The two MS methods performed here suggest altered arrangement of the extracellular matrix in Fuchs’ endothelial corneal dystrophy and provide new candidate proteins that may be involved in molecular mechanism of this disease.
2014-05-27 | PXD000746 | Pride
Project description:The in situ formation of adhesive hydrogels can repair of corneal defects but most focused on repairing focal stromal defects with diameters ≤ 3.5 mm because of their weak adhesion to wet tissues and fast biodegradation rate. Herein, a photocurable bioadhesive hydrogel that mimics the extracellular matrix (ECM) composition is developed for repairing 6-mm-diameter corneal stromal defects in rabbits. This biomacromolecule-based matrix hydrogel can undergo rapid gelling after UV light irradiation, exhibiting a high light transmittance and favorable mechanics. More importantly, this hydrogel maintained the viability and adhesion of cornea-derived cells and promoted their migration in vitro in 2D and 3D culture environments. Furthermore, in a rabbit corneal stromal defect model, the bioadhesive hydrogel could rapidly sealed wet corneal defects, resulting in epithelial wound coverage within 3-4 weeks, thereby contributing to a decrease in scar formation and increase in corneal stromal-neural regeneration during the 6-month follow-up. Thus, this work indicates that the biomacromolecule-based matrix hydrogel displays great potential for the regeneration of large-diameter corneal defects.
| PXD037052 |
Project description:Corneal injuries remain a major cause of consultation in the ophthalmology clinics worldwide. Repair of corneal wounds is a complex mechanism that involves cell death, migration, proliferation, differentiation, and extracellular matrix (ECM) remodeling. In the present study, we used a tissue-engineered, two-layers (epithelium and stroma) human cornea as a biomaterial to study both the cellular and molecular mechanisms of wound healing. Gene profiling on microarrays revealed important alterations in the pattern of genes expressed by tissue-engineered corneas in response to wound healing. Expression of many MMPs-encoding genes was shown by microarray and qPCR analyses to increase in the migrating epithelium of wounded corneas. Many of these enzymes were converted into their enzymatically active form as wound closure proceeded. In addition, expression of MMPs by human corneal epithelial cells (HCECs) was affected both by the stromal fibroblasts and the collagen-enriched ECM they produce. Most of all, results from mass spectrometry analyses provided evidence that a fully stratified epithelium is required for proper synthesis and organization of the ECM on which the epithelial cells adhere. In conclusion, and because of the many characteristics it shares with the native cornea, this human two layers corneal substitute may prove particularly useful to decipher the mechanistic details of corneal wound healing.
2015-11-25 | GSE75336 | GEO
Project description:The cornea is the transparent tissue at the forefront of the eye. Its outermost layer, the corneal epithelium, acts as a protective barrier for the ocular chamber. The cornea is avascular and its homeostasis relies on alternative sources to receive nutrients and growth factors, essential for maintaining its physiological functions and transparency. These sources include the corneal epithelium itself, the innervation and the tear film, collectively forming the corneal microenvironment. Any disruption to this finely balanced system (due to injury, ageing or diseases) can impair corneal transparency and lead to a progressive opacification, ultimately resulting in vision loss. Corneal blindness affects over 28 million people worldwide and represents the fourth leading cause of blindness. Among corneal injuries, abrasions are defined by a transient disruption in the integrity and cohesion of the epithelial barrier and are the most common type of eye injury encountered in clinical practice. Abrasion triggers a complex corneal wound healing process, involving the whole microenvironment, essential to restore transparency. However, a delayed or defective healing can lead to severe complications and visual impairments. Frequently caused by traumatic events, abrasion is also a mandatory step for some refractive surgeries such as photorefractive keratectomy (PRK). Understanding the molecular mechanisms underlying corneal wound healing is thus critical to improve the clinical outcomes and minimize the risk of complications such as haze or delayed epithelial regeneration. The aim of this study is to characterize the temporal dynamics of the tear fluid proteome following PRK and identify molecular signatures associated with corneal epithelial healing. We conducted a prospective cohort study involving longitudinal proteomic analysis of tear samples with mass spectrometry at baseline (preoperative), 30 minutes postoperatively (day 0), and on day 3 after PRK. Ten healthy adult patients undergoing bilateral PRK for myopia correction were included in this study. Corneal epithelial injury was induced by bilateral PRK, a standardized surgical model of epithelial removal followed by laser ablation.
2025-11-10 | PXD067691 | Pride