Project description:Purpose: To investigate the changes in gene expression induced by cyclic mechanical stress (CMS) in trabecular meshwork (TM) 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.
Project description:The purpose of this study was to understand the response of trabecular meshwork under mechanical stretch using a multi-omics approach.
Project description:Glucocorticoids with different chemical structures but similar glucocorticoid receptor potency regulate subsets of common and unique genes in human trabecular meshwork cells. Gene expression changes of human trabecular meshwork cells, TM 86 and TM 93, due to treatment with dexamethasone (Dex), fluocinolone acetonide (FA), and triamcinolone acetonide (TA).
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
