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Changes in Gene expression mediated by Cyclic Mechanical Stress in Trabecular Meshwork Cells

ABSTRACT: Purpose: To investigate the changes in gene expression induced by cyclic mechanical stress (CMS) in trabecular meshwork (TM) cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE14768 | GEO | 2009/03/04

SECONDARY ACCESSION(S): PRJNA112149

REPOSITORIES: GEO

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Changes in Gene expression mediated by Cyclic Mechanical Stress in Trabecular Meshwork Cells

Project description:Purpose: To investigate the changes in gene expression induced by cyclic mechanical stress (CMS) in trabecular meshwork (TM) cells. Methods: HTM cultures from three donors were plated on type I collagen-coated flexible silicone bottom plates and subjected to 15% stretching, 1 cycle per second for 6 hours. Non-stressed parallel control cultures were incubated under the same conditions in the absence of CMS. Total RNA from each culture was amplified (1 round amplification) and hybridized to Operon Human Oligo Arrays version 3.0 (35K). Control used was Universal Reference Human RNA from Stratagene. Differences in gene expression induced by CMS were analyzed using Genespring 7.2.

2009-03-04 | E-GEOD-14768 | biostudies-arrayexpress

Genome-wide Expression Profiling and Pathway Analysis in Cyclic Stretched Human Trabecular Meshwork Cells

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Project description:Microarray study comparing the trabecular meshwork (TM) with TM-MSC, corneal and scleral tissues

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Project description:Trabecular meshwork (TM) tissue is subjected to constant mechanical stress due to the ocular pulse created by the cardiac cycle. This induces alterations in membrane lipids and associated cell-cell adhesion and cell-extracellular matrix (ECM) interactions triggering intracellular signaling responses to counter mechanical insults. A loss of such response can lead to elevated intraocular pressure (IOP), a major risk factor for primary open-angle glaucoma. The purpose of this study was to understand the response of TM under mechanical stretch using a multi-omics approach. We performed an unbiased mRNA sequencing for changes in transcripts, mass spectrometry (MS)-based quantitative proteomics for protein changes, and multiple reaction monitoring (MRM) profiling based MS and HPLC-based MS to identify lipid changes. With the help of the multi-omics data analysis on human TM cells under mechanical stress, we provide evidence for the regulation of TM actin cytoskeleton and ECM interactions at the mRNA and protein levels, which can further modulate the TM lipids contents effectively regulating the mechanical properties of TM membrane. Overall, in this study, we propose the mechanistic interplay of macromolecules to bring about a concerted cellular response in TM cells to achieve mechanotransduction and IOP regulation

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Gene expression profile of human trabecular meshwork cells with silenced expression of autophagy genes Atg5 and Atg7 subjected to cyclic mechanical stretch

Project description:In order to investigate the effects of downregulated autophagy in TM cells response to mechanical stretch, we conducted gene expression analysis in human TM cells deficient in autophagy and subjected to cyclic mechanical stretch. For this, three independent strains of primary human TM cells were transfected with a cocktail of siRNAs to specifically silence the expression of the autophagy genes Atg5 and Atg7 (siAtg5/7), and subjected to cyclic mechanical stress (15% elongation, 1 cycle/sec, 24h).

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Project description:Treatment with glucocorticoids is known to cause ocular hypertension in humans which can lead to steriod-induced glaucoma We used microarrays to determine changes in gene expression in two human trabecular meshwork (TM) cell isolates (TM-4 and TM-2) from the same donor that could explain the increase in ocular hypertension.

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Project description:The trabecular meshwork (TM) responsible for IOP homeostasis in the eye could sense the IOP fluctuations dynamically and adapt to the mechanical changes appropriately. Cationic mechanosensitive channels (CMCs) have been reported with critical roles in mediating the TM responding to many mechanical forces. However, the mechanism of how CMCs influence the TM cellular function in aqueous humor drainage and against mechanical damage is still elusive. Gene ontology enrichment analysis demonstrated that prohibition of CMCs significantly influenced the mechanical changes in the TM, such as store-operated calcium channel activity, microtubule cytoskeleton polarity, toll-like receptor signaling pathway, and neuron cell fate specification. Our results indicated that they might be the critical downstream signals of CMCs adapting to mechanical forces and mediating AH outflow.

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