Genome-Wide RNA Sequencing of Human Trabecular Meshwork Cells Treated with TGF-β1: Relevance to Pseudoexfoliation Glaucoma

Abstract

Pseudoexfoliation glaucoma (XFG) is an aggressive form of secondary open angle glaucoma, characterised by the production of exfoliation material and is estimated to affect 30 million people worldwide. Activation of the TGF-β pathway by TGF-β1 has been implicated in the pathogenesis of pseudoexfoliation glaucoma. To further investigate the role of TGF-β1 in glaucomatous changes in the trabecular meshwork (TM), we used RNA-Seq to determine TGF-β1 induced changes in the transcriptome of normal human trabecular meshwork (HTM) cells. The main purpose of this study was to perform a hypothesis-independent RNA sequencing analysis to investigate genome-wide alterations in the transcriptome of normal HTMs stimulated with TGF-β1 and investigate possible pathophysiological mechanisms driving XFG. Our results identified multiple differentially expressed genes including several genes known to be present in exfoliation material. Significantly altered pathways, biological processes and molecular functions included extracellular matrix remodelling, Hippo and Wnt pathways, the unfolded protein response, oxidative stress, and the antioxidant system. This cellular model of pseudoexfoliation glaucoma can provide insight into disease pathogenesis and support the development of novel therapeutic interventions.

Genome-wide RNA sequencing of ocular fibroblasts from glaucomatous and normal eyes: Implications for glaucoma management

“…In keeping with our results, metallopeptidase ADAM12 has also been reported to be upregulated in glaucomatous lamina cibrosa cells [ 53 ] and is also associated with liver-fibrosis [ 54 ]. The expression of ADAM12 is regulated by TGF-β1 [ 55 ] and expression was shown to be upregulated in vitro in human trabecular meshwork cells after treatment with TGF-β1 [ 56 ]. Several lysyl oxidases were upregulated in our results which has also been shown in other myofibroblasts [ 34 ].…”

Exploring Epigenetic Modifications as Potential Biomarkers and Therapeutic Targets in Glaucoma

“…Another factor causing the increase in TM fibrosis is given as the demethylation of the GDF7 promoter which induces the transcription of the growth differentiation factor (GDF7) gene, a member of the TGFβ superfamily [ 22 , 23 , 24 ]. In rhesus macaque models, TM fibrosis was inhibited by antibodies directed against GDF7.…”

The Hippo signalling pathway and its impact on eye diseases

“…Genome-wide transcriptome analysis of HTM cells treated with TGF-β1/2 showed that the Hippo signalling pathway is related to POAG and TM pathophysiology. 95,96 YAP/TAZ are present in the TM, and YAP functions in normal trabecular tissue, whereas TAZ has a major impact on glaucomatous trabecular tissue. 97 Both matrix stiffness 91 and composition 98 can affect YAP/TAZ activation in HTM.…”