Abstract
Fibrosis-related events play a part in most blinding diseases worldwide. However, little is known about the mechanisms driving this complex multifactorial disease. Here we have carried out the first genome-wide RNA-Sequencing study in human conjunctival fibrosis. We isolated 10 primary fibrotic and 7 non-fibrotic conjunctival fibroblast cell lines from patients with and without previous glaucoma surgery, respectively. The patients were matched for ethnicity and age. We identified 246 genes that were differentially expressed by over two-fold and p < 0.05, of which 46 genes were upregulated and 200 genes were downregulated in the fibrotic cell lines compared to the non-fibrotic cell lines. We also carried out detailed gene ontology, KEGG, disease association, pathway commons, WikiPathways and protein network analyses, and identified distinct pathways linked to smooth muscle contraction, inflammatory cytokines, immune mediators, extracellular matrix proteins and oncogene expression. We further validated 11 genes that were highly upregulated or downregulated using real-time quantitative PCR and found a strong correlation between the RNA-Seq and qPCR results. Our study demonstrates that there is a distinct fibrosis gene signature in the conjunctiva after glaucoma surgery and provides new insights into the mechanistic pathways driving the complex fibrotic process in the eye and other tissues.
Highlights
Some of the authors of this publication are working on these related projects: Nanoparticles for topical administration of siRNA to the airways for cystic fibrosis therapy View project
The MYOCD gene encodes myocardin, a smooth muscle-specific transcriptional co-activator of serum response factor (SRF), and its expression was significantly upregulated in fibrotic fibroblast (FFs) compared to non-fibrotic fibroblast (NFs) (Table S1)
In the disease association analysis, the LMO3, MYB and BIRC3 oncogenes were significantly upregulated whereas the WISP2, Insulin-like growth factor-binding protein 5 (IGFBP5) and RASSF2 tumour suppressor genes were significantly downregulated in FFs compared to NFs
Summary
Some of the authors of this publication are working on these related projects: Nanoparticles for topical administration of siRNA to the airways for cystic fibrosis therapy View project. All content following this page was uploaded by Cynthia Yu-Wai-Man on 18 July 2017. Our study demonstrates that there is a distinct fibrosis gene signature in the conjunctiva after glaucoma surgery and provides new insights into the mechanistic pathways driving the complex fibrotic process in the eye and other tissues. Identifying the dysregulated genes will be important in understanding the complex network of signalling pathways driving the fibrotic process and in finding potential novel therapeutic targets and biomarkers of disease severity and prognosis in fibrotic eye diseases. RNA-Sequencing (RNA-Seq) is a powerful and highly sensitive technology that allows the whole transcriptome to be studied compared to DNA microarrays and requires smaller amounts of RNA5, 6
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