Long non-coding RNA HOTAIR induces GLI2 expression through Notch signalling in systemic sclerosis dermal fibroblasts
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(2020) 22:286
RESEARCH ARTICLE
Open Access
Long non-coding RNA HOTAIR induces GLI2 expression through Notch signalling in systemic sclerosis dermal fibroblasts Christopher W. Wasson1, Rebecca L. Ross1, Rebecca Wells1,2, Clarissa Corinaldesi1, Ioanna Ch. Georgiou2, Natalia A. Riobo-Del Galdo2,3 and Francesco Del Galdo1,4*
Abstract Objectives: Systemic sclerosis (SSc) is characterised by tissue fibrosis of the major organs of the body including the skin, lungs and heart. We have previously reported that the lncRNA HOTAIR plays a central role in the activation of SSc myofibroblasts, the key cellular elements of fibrosis. HOTAIR induces fibroblast activation through H3K27me3mediated activation of the Notch signalling pathway. Here we aimed to identify the signalling events downstream of Notch that drive SSc myofibroblast activation. Methods: Patient fibroblasts were obtained from full-thickness forearm skin biopsies of 3 adult patients with SSc of recent onset. The lncRNA HOTAIR was expressed in healthy dermal fibroblasts by lentiviral transduction. Hedgehog signalling pathway was inhibited with GANT61 and GLI2 siRNA. Gamma secretase inhibitors RO4929097 and DAPT were used to block Notch signalling. GSK126 was used to inhibit Enhancer of Zeste 2 (EZH2). Results: Overexpression of HOTAIR in dermal fibroblasts induced the expression of the Hedgehog pathway transcription factor GLI2. This is mediated by activation of Notch signalling following epigenetic downregulation of miRNA-34a expression. Inhibition of H3K27 methylation and Notch signalling reduced expression of GLI2 in HOTAIRexpressing fibroblasts as well as in SSc dermal fibroblasts. Importantly, the inhibition of GLI2 function using GANT61 or siRNA mitigates the pro-fibrotic phenotype induced by HOTAIR. Conclusions: Our data indicates that GLI2 expression is stably upregulated in SSc myofibroblasts through HOTAIR and that GLI2 mediates the expression of pro-fibrotic markers downstream of Notch. Keywords: Systemic sclerosis, Hedgehog signalling, Epigenetics, Long non-coding RNA, HOTAIR
Introduction Systemic sclerosis (SSc) is an autoimmune condition that initially presents in the skin of the patient’s hands and feet, where there is a build-up of extracellular matrix resulting in skin fibrosis. The disease progresses to the forearms and legs and in the most severe cases (diffuse * Correspondence: [email protected] 1 Leeds Institute of Rheumatic and Musculoskeletal Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, UK 4 Scleroderma Programme, NIHR Leeds Musculoskeletal Biomedical Research Centre, Leeds, UK Full list of author information is available at the end of the article
SSc) to the trunk. Beyond skin fibrosis, SSc can cause tissue fibrosis in the internal organs (lungs, heart and kidneys). The key role of tissue fibroblasts in the disease has been well-characterised. A number of signalling pathways such as Transforming Growth Factor beta (TGF-β) [1–3], Sonic Hedgehog (Shh) [4–6] and Notch signalling [7, 8] have been implicated in d
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