Project description:Long noncoding RNAs (lncRNAs) are emerging as important regulators in cellular processes and have been showed to be involved in the occurrence and development of various neurodegenerative diseases including glaucoma. The aim of this study is to reveal disease-related extracellular lncRNAs and message RNAs (mRNAs) in aqueous humor (AH) of individual primary open-angle glaucoma (POAG) patients, to determine the potential biomarkers for POAG diagnosis
Project description:To better understand the molecular changes in the aqueous humor (AH) content with glaucoma, we analyzed the microRNA (miRNA) profiles of AH samples from patients with Primary Open Angle Glaucoma (POAG) and Exfoliation Glaucoma (XFG) compared to non-glaucoma controls.
Project description:TGF-beta levels are known to increase in the aqueous humor of eye cells in patients with glaucoma. Increase TGF-beta is assumed to have a biochemical impact on the trabecular meshwork, and an increase in extracellular matrix formation, which may be responsible for decrease outflow facility of the eye. This may increase extracellular pressure, causing glaucoma. TGF-beta 1 may be the cause of abnormal accumulation of extracellular matrices in trabecular meshwork of eyes with primary open angle glaucoma. Transforming growth factor (TGF)-beta2 regulates the expression of proteoglycans in aqueous humor from human glaucomatous eyes. To identify gene expression changes as a result of TGF-beta1 and 2 treatment of human trabecular meshwork cells. We expect to see a change in expression of the proteoglycans in HTM cells as a response to TGF-beta treatment. Human Trabecular Meswork cells in the eye were bathed by aqueous humor. TM cells were removed from individuals with the following ages: 16,66,67,73, and 76. Each individual was treated with EtOH (control), TGF-beta1, or TGF-beta2. Total RNA from each individual was pooled for each chip. Technical replicates were created for each treatment type, for a total of 6 chips.
Project description:TGF-beta levels are known to increase in the aqueous humor of eye cells in patients with glaucoma. Increase TGF-beta is assumed to have a biochemical impact on the trabecular meshwork, and an increase in extracellular matrix formation, which may be responsible for decrease outflow facility of the eye. This may increase extracellular pressure, causing glaucoma. TGF-beta 1 may be the cause of abnormal accumulation of extracellular matrices in trabecular meshwork of eyes with primary open angle glaucoma. Transforming growth factor (TGF)-beta2 regulates the expression of proteoglycans in aqueous humor from human glaucomatous eyes. To identify gene expression changes as a result of TGF-beta1 and 2 treatment of human trabecular meshwork cells. We expect to see a change in expression of the proteoglycans in HTM cells as a response to TGF-beta treatment. Human Trabecular Meswork cells in the eye were bathed by aqueous humor. TM cells were removed from individuals with the following ages: 16,66,67,73, and 76. Each individual was treated with EtOH (control), TGF-beta1, or TGF-beta2. Total RNA from each individual was pooled for each chip. Technical replicates were created for each treatment type, for a total of 6 chips. Keywords: dose response
Project description:Globally, irreversible vision loss is mainly attributed to glaucoma, a complex eye condition. Given that the progression of glaucoma is challenging to detect and track, and reliable predictive markers are lacking, there is an urgent need for biomarkers. In this context, the aqueous humor, which is part of the anterior segment of the eye, assumes a critical role. The aim of this study is to develop a comprehensive database for glaucoma research by analyzing primary open-angle glaucoma (POAG) human aqueous humor samples (n=66). We employ advanced label-free quantitative proteomics to investigate the proteome associated with glaucoma in human aqueous humor. Our dataset, generated using the TimsTOF™ Pro Bruker mass spectrometer and DIA-NN bioinformatics for library-independent quantification of data-independent acquisition (DIA) proteomics, offers researchers access to the most extensive catalog of human aqueous humor proteins reported to date. This resource is expected to deepen our understanding of the proteomic profiles in human glaucoma aqueous humor and also support investigations into proteomes related to other ocular conditions.
Project description:Long non-coding RNAs were associated with the development and progression of glaucoma. Our study aim to identify the potential genes in human trabecular meshwork related to primary open-angle glaucoma (POAG).
Project description:Comparative evaluation of the aqueous humor proteome of primary angle closure and primary open angle glaucomas and senile cataract eyes
Project description:A high percentage of vision loss and blindness worldwide are due to glaucoma and cataracts ocular diseases. Small extracellular vesicles (sEVs) are small lipidic vesicles involved in transport and cell-to-cell communication, released into different body fluids including eye’s aqueous humor. The identification and characterization of the protein content of small extracellular vesicles in ocular pathologies has not been well elucidated yet. To that end, a by quantitative proteomics approach using Tandem Mass Tag (TMT) 10-Plex was performed to analyze the protein content of extracellular vesicles from patients´ aqueous humors with cataracts and glaucoma compared to healthy individuals (ICL). Labeled peptides were analyzed on an Orbitrap Exploris 480 equipped with FAIMS Pro Duo interface. Then, western blot (WB) and ELISA analyses using aqueous humor samples from ICL, cataracts, and glaucoma patients were performed for validation. From the TMT 10-plex analysis of sEVs, a total of 828 peptides and 192 proteins were identified and quantified. After data analysis with R program, 14 significantly dysregulated proteins from sEVs of aqueous humor in cataracts and 11 in glaucoma showed a fold change ≥1.5. Then, the dysregulation of 10 candidate dysregulated proteins was confirmed by WB and ELISA using directly aqueous humor samples. In addition, GAS6 and SPP1 were found to possess a high diagnostic ability of glaucoma patients. In conclusion, the proteomics analysis of sEVs from aqueous humor samples allowed the identification of GAS6 and SPP1 as glaucoma biomarkers, which in combination allowed for discriminating glaucoma patients from control individuals with an area under the curve (AUC) of 76.1%, and a sensitivity of 65.6%, and a specificity of 87.7%.
