Influence of cyclical mechanical strain on global gene expression in human lamina cribrosa cells in vitro
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE3166 | GEO | 2005/08/21
SECONDARY ACCESSION(S): PRJNA93007
REPOSITORIES: GEO
Similar Datasets
Project description:The mechanical effect of raised intraocular pressure is a recognised stimulus for optic neuropathy in primary open angle glaucoma (POAG). Characteristic extra-cellular matrix (ECM) remodelling accompanies axonal damage in the lamina cribrosa (LC) of the optic nerve head in POAG. Glial cells in the lamina cribrosa may play a role in this process but the precise cellular responses to mechanical forces in this region are unknown. The authors examined global gene expression profiles in lamina cribrosa cells exposed to cyclical mechanical stretch, with an emphasis on ECM genes. Experiment Overall Design: n=3 stretch and static control experiments. Experiment Overall Design: RNA pooled from each experiment and hybridised to individual experiment and control arrays.
2008-06-12 | E-GEOD-3166 | biostudies-arrayexpress
Project description:Primary open angle glaucoma (POAG) is a progressive optic neuropathy, which is a major cause of worldwide visual impairment and blindness. Pathological hallmarks of the glaucomatous optic nerve head include retinal ganglion cell axon loss and extracellular matrix (ECM) remodeling of the lamina cribrosa layer. TGF-beta is an important pro-fibrotic modulator of ECM metabolism, whose levels are elevated in human POAG lamina cribrosa tissue compared with non-glaucomatous controls. We treated human lamina cribrosa (LC) cells with TGF-beta1 (10ng/ml) for 24 hours in order to examine differential gene expression patterns in repsonse to this cytokine. In particular we focused on ECM and fibrotic genes. We found that TGF-beta1 induces expression and release of ECM components in LC cells, which may be important in regulating matrix remodeling in the lamina cribrosa. In disease states such as POAG, the LC cell may represent an important pro-fibrotic cell type and an attractive target for novel therapeutic strategies.
2008-06-12 | E-GEOD-2705 | biostudies-arrayexpress
Project description:Primary open angle glaucoma (POAG) is a progressive optic neuropathy, which is a major cause of worldwide visual impairment and blindness. Pathological hallmarks of the glaucomatous optic nerve head include retinal ganglion cell axon loss and extracellular matrix (ECM) remodeling of the lamina cribrosa layer. TGF-beta is an important pro-fibrotic modulator of ECM metabolism, whose levels are elevated in human POAG lamina cribrosa tissue compared with non-glaucomatous controls. We treated human lamina cribrosa (LC) cells with TGF-beta1 (10ng/ml) for 24 hours in order to examine differential gene expression patterns in repsonse to this cytokine. In particular we focused on ECM and fibrotic genes. We found that TGF-beta1 induces expression and release of ECM components in LC cells, which may be important in regulating matrix remodeling in the lamina cribrosa. In disease states such as POAG, the LC cell may represent an important pro-fibrotic cell type and an attractive target for novel therapeutic strategies. Keywords: other
2005-08-05 | GSE2705 | GEO
Project description:Purpose: Marked extracellular matrix (ECM) remodeling occurs in the human optic nerve head in primary open angle glaucoma (POAG). The glial fibrillary acid protein (GFAP) negative lamina cribrosa cell may play an important role in this remodeling process. The authors report the first study of global and ECM-focused gene transcription differentials between GFAP-negative negative lamina cribrosa (LC) cells from normal and POAG human donors. Methods: GFAP-negative LC cell lines were generated from the optic nerve tissue of three normal (n=3) and three POAG (n=3) human donors. Using Affymetrix U133A arrays the transcriptional profile between the normal and diseased groups were compared. Bioinformatic analysis was carried out using robust multichip average (RMA Express) and EASE/David. Real time TaqMan PCR and immunohistochemistry analyses were performed to validate the microarray data. Results: 285 genes were up regulated by greater than 1.5 fold and 413 were down regulated by greater than 1.5 fold in the POAG LC cells versus normal controls. Upregulated genes in POAG LC cells included, SPARC, periostin, thrombospondin, CRTL-1, CTGF and collagen types I, III, V and VIII. Downregulated ECM genes in POAG included MMP-1, fibulin, decorin and tenacsin XB. All TaqMan PCR validation assays were significant (*p<0.05) and consistent with the array data. Immunohistochemistry of one target (periostin) confirmed its differential expression at the protein level in POAG optic nerve head tissue compared with non-glaucomatous controls. Functional annotation and over-representation analysis identified ECM genes as a statistically over-represented class of genes in POAG LC cells compared with normal LC cells. Conclusions: This study reports for the first time that POAG LC cells in-vitro demonstrate up regulated ECM and pro-fibrotic gene expression compared with normal LC cells. This may be a pathological characteristic of this cell type in POAG in-vivo. We believe that the LC cell may be a pivotal regulator of optic nerve head ECM remodeling and an attractive target for future therapeutic strategies in POAG.
2009-01-06 | GSE13534 | GEO
Project description:Primary Open-Angle Glaucoma (POAG) and Pseudoexfoliation Glaucoma (PEXG) are forms of glaucoma with distinct etiologies, but they share a common type of optic neurodegeneration that leads to irreversible loss of retinal ganglion cells, visual field and blindness if left untreated. The main objective in the present study was to explore whether differential-proteomic analysis of blood serum from patients with POAG or PEXG could lead to the identification of candidate biomarkers of glaucoma and provide clues to pathogenic mechanisms. A total of 202 blood samples from medically treated patients with POAG (n=73), PEXG (n=59), and cataracts (control, n=70) were collected and analyzed following a proteomic workflow that involved two phases: discovery and validation. In the discovery phase, differential-proteomics (2D-DIGE) methodology led to the identification of a panel of 35 proteins that were altered (i.e., over- or under-expressed) in serum samples of glaucoma patients. Functional pathway analysis suggested a potential involvement of these proteins in immunological and inflammatory pathways. In the validation phase, ELISA assays of the top-17-ranked proteins confirmed that most of them were over-expressed in the glaucoma groups. Stepwise discriminant analysis of the ELISA data resulted in a six-protein panel (APOA4, TF, C3, APOL1, IGHG2, C4A) that was used to generate multivariate predictive models. These models achieved a diagnostic efficiency of 81% (correct assignment), specificity of 83%, and sensitivity of 92%. Specifically, 100% efficacy was reached in the discrimination of the POAG group from control, and 86% efficacy in the discrimination of POAG from PEXG. Overall, these data demonstrate the discovery of a panel of serum biomarkers that could be used in large-scale multiplexed screenings for the diagnosis of POAG and PEXG patients. Furthermore, these biomarkers may have a role in the pathogenesis, and association to immune-inflammatory processes with glaucoma.
2014-01-17 | PXD000198 | Pride
Project description:Influence of cyclical mechanical strain on global gene expression in human lamina cribrosa cells in vitro
| PRJNA93007 | ENA
Project description:Glaucoma is a progressive optic neuropathy that can lead to irreversible blindness. Its main risk factor is elevated intraocular pressure (IOP). The trabecular meshwork (TM) acts as a filter between the anterior chamber of the eye and the aqueous humor collecting ducts, and dysfunction of this meshwork is responsible for the increased IOP in primary open-angle glaucoma (POAG). Considering that the culture conditions of human TM cells (HTMC) influence gene expression, we used human TM explants (HTMEx), which most closely mimic physiological conditions, to study the transcriptome of HTMC. The transforming growth factor-beta 2 (TGF-β2) signaling pathway has been implicated in the pathophysiology of POAG. To better characterize the role of TGF-β2 in this pathophysiology, we used bulk RNA sequencing and immunohistological analyses to establish gene signatures of TGF-β2-exposed HTMEx and correlate them with morphological alterations. We identified differentially upregulated genes primarily involved in ECM regulation, as well as profibrotic TGF-β signaling pathways, confirmed using confocal microscopy to highlight changes in trabecular architecture, TGFβ2-induced F-actin rearrangements, and extracellular matrix (ECM) deposition. Enrichment analysis also revealed modulations of gene expression related to cytoskeletal organization, as well as activation of the bone morphogenic protein (BMP) and Wnt signaling pathways in response to TGF-β2.
2023-07-03 | GSE236302 | GEO
Project description:Glaucoma is a group of diseases characterized by progressive optic nerve damage leading to irreversible visual field loss, often associated with elevated intraocular pressure (IOP). However, the specific underlying factors or molecular mechanisms distinguishing various glaucoma subtypes are still not fully understood. In this study, we enrolled a total of 27 subjects, comprising 5 exfoliation syndrome (XFS), 4 exfoliation glaucoma (XFG), 11 primary open-angle glaucoma (POAG), and 7 cataract patients (Control) for a comparative multi-omics analysis including both proteomics and metabolomics of the aqueous humor (AH). The AH proteome and metabolome were analyzed separately using data-independent acquisition (DIA) and data-dependent acquisition (DDA) methods, after which the results were additionally integrated for a comprehensive analysis. The comparative proteomics analysis led to the identification of differentially expressed proteins (DEPs) related to lipid metabolism, complement activation, and extracellular matrix (ECM) regulation, which were commonly up-regulated in both XFG and POAG groups. The metabolomics analysis revealed differentially expressed metabolites (DEMs) particularly those involved in amino acids, suggesting their role in antioxidative processes. Thus, bioinformatic analysis suggested significant substances, such as VTN, APOA1, C6, and L-phenylalanine, that showed quantitative alterations in glaucoma patients. In addition, the integrating the individual omics analyses revealed interaction networks involving PLG, APOA1, and L-phenylalanine, or C3, APOD, and L-valine, suggesting the quantitative relationship in protein-metabolite interactions related to inflammation and lipid metabolism within the AH of glaucoma patients. Furthermore, the correlation approach identified a relationship between FGG and L-phenylalanine, and VTN and inosine, implying potential interactions among compounds not previously identified. These findings would provide valuable insights into the complex molecular mechanism differentiating glaucoma subtypes and may benefit further research into the clinical application of AH proteome and metabolome.
2024-04-11 | PXD046219 | Pride
Project description:We determined the differential expression of miRNAs that target extracellular matrix mRNAs in glauocmatous and TGFb2 treated lamina cribrosa cells compared to normal lamina cribrosa cells.
2021-06-15 | GSE178137 | GEO
Project description:Although cells of the immune system experience force and pressure throughout their lifecycle, almost nothing is known about how these mechanical processes regulate the immune response. Immune cells in highly mechanical organs, such as the lung, are constantly exposed to tonic and dynamically changing mechanical cues. Here using reverse genetics, we show that myeloid cells respond to force and alterations in cyclical hydrostatic pressure (CHP) via the mechanosensory ion channel (MSIC) PIEZO1. Unbiased RNA-sequencing from macrophages subjected to CHP reveals a striking state of proinflammatory reprogramming. We report a novel mechanosensory-immune signaling circuit which PIEZO1 initiates in response to CHP, activating c-JUN, upregulating Endothelin-1 (EDN1), and stabilizing HIF1α to facilitate a prolonged program of proinflammatory mediators. Using mice conditionally deficient of PIEZO1 in myeloid cells, and cellular depletion assays, we show infiltrating monocytes respond to cyclical force to recruit neutrophils and clear pulmonary Pseudomonas aeruginosa infection. Furthermore, myeloid PIEZO1 also drove lung pathology in a mouse model of pulmonary fibrosis. Our results demonstrate a novel environmental sensory axis that myeloid cells recognize to mount an inflammatory response, and is the first report showing a physiological role for PIEZO1 and mechanosensation in immunity.
2019-08-21 | GSE133069 | GEO