Transcriptomic analysis of TGFβ-mediated fibrosis in primary human Tenon’s fibroblasts

Author:

Pasvanis Zoe1,Boynes Antony1,Kong Roy C.K.1,Chan Elsa C.1,Wong Raymond C.B.1,Gaskin Jennifer Fan1

Affiliation:

1. Centre for Eye Research Australia

Abstract

Abstract

Glaucoma filtration surgery (GFS) is performed to slow down disease progression in glaucoma, a leading cause of irreversible blindness worldwide. Following surgery, pathological wound healing may lead to conjunctival fibrosis and filtering failure. Myofibroblasts are the key cells responsible for postoperative conjunctival scarring. This study aims to further understand the molecular mechanisms of conjunctival fibrosis following GFS. We utilised RNA-sequencing (RNA-seq) to delineate the TGFβ1 induced changes in the transcriptome of human Tenon’s fibroblasts (HTFs). RNA sequencing was performed on HTFs after 5 days of TGFβ1 treatment. Following quality control, 3,362 differentially expressed genes were identified, of which 1,532 were upregulated and 1,820 were downregulated. We identified signaling pathways associated with the pathogenesis of conjunctival fibrosis. The DEGs (differentially expressed genes) were enriched in pathways including myofibroblast differentiation, TGFβ-signaling, collagen and extracellular matrix organization, epithelial to mesenchymal transition, and cell cycle regulation. The results of this study identified the transition from HTF to myofibroblast is characterised by the upregulation of key genes including LDLRAD4, CDKN2B, FZD8, MYOZ1, and the downregulation of SOD3, LTBP4 and RCAN2. This insight into the transcriptional landscape of HTFs and myofibroblast differentiation is essential to understand the pathophysiology of conjunctival scarring and develop new therapeutic agents.

Publisher

Springer Science and Business Media LLC

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