Integrated analysis of transcriptome and proteome of the human cornea and aqueous humor reveal novel biomarkers for corneal endothelial cell dysfunction
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ABSTRACT:
PROVIDER: PRJNA736411 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14768214_1.fastq.gz | Fastqsanger.gz | |||
| SRR14768214_2.fastq.gz | Fastqsanger.gz | |||
| SRR14768215_1.fastq.gz | Fastqsanger.gz | |||
| SRR14768215_2.fastq.gz | Fastqsanger.gz | |||
| SRR14768216_1.fastq.gz | Fastqsanger.gz |
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Project description:Previous studies have reported that elevated protein levels in the aqueous humor (AH) are associated with corneal endothelial cell dysfunction (CECD), but the detailed underlying mechanism as well as specific biomarkers for CECD remain elusive. In the present study, we aimed to identify protein markers in AH directly associated with changes in Corneal endothelial cells (CECs), as AH can be easily obtained for analysis. We carried out an in-depth proteomic analysis of patient-derived AH as well as transcriptomic analysis of CECs from the same patients with bullous keratopathy (BK) resulting from CECD. We first determined differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) from CECs and AH in CECD, respectively. By combining transcriptomic and proteomic analyses, 13 shared upregulated markers were observed between DEGs and DEPs. Through data integration with individual analysis via data-independent acquisition, three upregulated transcripts and proteins were identified, namely metallopeptidase inhibitor 1 (TIMP1), Fc fragment of IgG binding protein (FCGBP), and angiopoietin-related protein 7 (ANGPTL7). Furthermore, we confirmed AH biomarkers for CECD using individual proteome verification and immunoassay validation. Conclusively, our findings may provide valuable insights into the disease process and identify biofluid markers for the assessment of CEC function during BK development.
2021-06-10 | GSE176481 | GEO
Project description:Although pig-to-nonhuman primate (NHP) corneal xenotransplantation has shown long-term graft survival, xenogeneic antigenrelated immune responses are still stronger than allogenic antigen-associated responses. Therefore, there is an unmet need to investigate major rejection pathways in corneal xenotransplantation, even with immunosuppression. This study aimed to identify biomarkers in aqueous humor for predicting rejection and to investigate rejection-related pathways in grafts from NHPs transplanted with porcine corneas following administration of steroids combined with tacrolimus/rituximab. NHPs who had received corneas from wild‐type (WT) or α‐1,3‐galactosyltransferase gene‐knockout (GTKO) pigs were divided into groups with or without rejection according to clinical examinations. Liquid chromatography-mass spectrometry (LC-MS) was used to analyze the proteomes of corneal tissues or aqueous humor. The biological functions of differentially expressed proteins (DEPs) were assessed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) for pathways and protein–protein interaction network analysis. Among the 66 DEPs in aqueous humor, complement proteins (C3, C5, and C9) and cholesterol metabolic proteins (APOA1 and APOA2) were related to xenogeneic rejection as biomarkers, and alternative pathways of the complement system seemed to be important in xenogeneic graft rejection. Among 416 DEPs of the cornea, NF‐κB1 and proteosomes (PSMD7, PSMA5, and PSMD3) seemed to be related with xenogeneic graft rejection. Additionally, oxidative phosphorylation and leukocyte activation‐related pathways are involved in rejection. Overall, our proteomic approach highlights the important role of NF‐κB1, proteosomes, oxidative phosphorylation, and leukocyte activation-related inflammation in the cornea, and the relevance of complement pathways of the aqueous humor as a predictive biomarker of xenogeneic rejection.
2022-05-20 | PXD031320 | Pride
Project description:INTRODUCTION: Cornea is the outermost part of the eye supplied mostly by aqueous humor (AH). Therefore, the comparison of the metabolomic compositions of AH and cornea may help to determine which compounds are produced inside the cornea, and which penetrate into cornea from AH for intra-corneal consumption. Keratoconus (KC) is the most common form of the cornea dystrophy, and the analysis of KC corneas can unravel the metabolomic changes occurring in AH and cornea of KC patients. </br> OBJECTIVES: The work is aimed at the determination of concentrations of a wide range of metabolites in the human cornea and AH, the comparison of the metabolomic profiles of cornea and AH, and the comparison of the metabolomic compositions of samples taken from KC patients and normal donors (post-mortem). </br> METHODS: The quantitative metabolomic profiling was carried out with the use of two independent methods—high-frequency 1H NMR spectroscopy and HPLC with high-resolution ESI-MS detection. </br> RESULTS: The concentrations of 71 most abundant metabolites in cornea and AH from keratoconus patients and from human cadavers have been measured. It is found that the concentrations of purines and organic acids in cornea are significantly higher than in AH. The KC corneas are characterized by the enhanced levels of acetate and citrate, and also by low values of GSH/GSSG ratios. </br> CONCLUSION: A significant difference in the metabolomic compositions of the human AH and cornea has been revealed. The concentrations of glucose and some amino acids in cornea are significantly lower than in AH, indicating their fast consumption inside the cornea. The high levels of organic acids, purines and GSH in cornea should be attributed to their production in the cornea. The enhanced levels of acetate and citrate as well as the low values of GSH/GSSG ratios in KC corneas are the indicators of the oxidative stress. </br>
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Project description:Purpose: To characterize microRNAs (miRNAs) and their possible roles in high myopia by using next generation sequencing Methods: Aqueous humor samples were obtained from 15 highly myopic eyes and 15 cataract eyes at the onset of surgery. miRNA next generation sequencing and bioinformatics analyses were performed using RNA extracted from aqueous humor samples. Results: A total of 341 miRNAs were detected in the aqueous humor samples of highly myopic eyes; 201 miRNAs were detected in the aqueous humor samples of cataractous control eyes. A total of 249 mature miRNAs and 17 novel miRNAs were differentially expressed during myopia. Possible pathways regulated by these aberrantly expressed miRNAs included the TNF, MAPK, PI3K-Akt, and HIF-1 signaling pathways. Conclusions: The current study provided an overall view of miRNA profiling in the aqueous humor of highly myopic eyes. These profiles may be associated with myopia pathogenesis, and are potential biomarkers.
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Project description:Aqueous humor (AH) is the fluid in the anterior and posterior chambers of the eye that contains proteins regulating ocular homeostasis. Analysis of aqueous humor proteome is challenging mainly due to low sample volume and protein concentration. In this study, by utilizing state of the art technology, we performed Liquid-Chromatography Mass spectrometry (LC-MS/MS) analysis of 88 aqueous humor samples from subjects undergoing cataract surgery.
2021-09-09 | PXD022463 | Pride
Project description:The purpose of this study is to discover genes that might increase aqueous humor outflow when human ciliary muscle or human trabecular meshwork cells are treated with the prostaglandin analogues latanoprost free acid or prostaglandin F2alpha. Five tissue donors were pooled on each chip. Keywords: other
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Project description:Identify DNA methylation change in retinoblastoma using aqueous humor cfDNA
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Project description:The aqueous humor is a colourless, transparent fluid that fills the anterior chamber of the eye. It has an important role to play in maintaining the intraocular pressure of the eye and providing nourishment to avascular structures such as the lens and cornea. It is also essential for providing a clear path for the entry and exit of light from the eye and maintaining the refractive power of the eye. In this study, 250 samples of aqueous humor were collected from patients undergoing cataract surgery. The samples were pooled, divided into aliquots and subjected to ingel digestion, insolution digestion followed by strong cation exchange chromatography and basic reverse phase liquid chromatography fractionation. MS/MS analysis was carried out on a Fourier transform LTQ-Orbitrap velos mass spectrometer and the data were searched with the search algorithms Mascot, SEQUEST and X!Tandem against the NCBI RefSeq human protein database. We have identified 818 proteins of which 447 have been identified for the first time. Proteins such as sorbitol dehydrogenase, filensin and phakinin were some of the novel proteins identified in this study. Classification of proteins based on domain information yielded 304 proteins with signal peptides, 46 proteins with transmembrane domain and 75 proteins with both signal peptide and transmembrane domains. Our study provides a detailed profile of the human aqueous humor. The results from this study of aqueous proteins should help facilitate research into pathological conditions of the eye such as glaucoma, myopia, cataract and uveitis.
2016-01-04 | PXD000787 | Pride