Suberanilohydroxamic acid (SAHA) inhibits collagen deposition in a transforming growth factor β1-driven precision cut lung slice (PCLS) model of pulmonary fibrosis

Brand, Oliver J.; Pasini, Alice; Habgood, Antony; Knox, Alan J.; Jenkins, Gisli; Pang, Linhua

Abstract

Introduction and Objectives: Idiopathic Pulmonary Fibrosis (IPF) is a chronic, progressive interstitial lung disease that is refractory to current treatment options. Transforming growth factor (TGF)-β1 is a key pro-fibrotic cytokine that plays a crucial role in IPF pathogenesis. Our group previously demonstrated distinct epigenetic modifications involved in repression of the antifibrotic gene cyclooxygenase-2 (COX-2) in fibroblasts from IPF (F-IPF) lungs compared with fibroblasts from non-fibrotic lungs (F-NL). Epigenetic drugs capable of inhibiting DNA and histone modifications may, therefore, represent a putative novel therapy. The aim of this study was to investigate the ability of 4 epigenetic inhibitors to regulate TGF-β-driven fibrosis in ex vivo mouse lung.
Methods: A precision-cut lung slice (PCLS) model of fibrosis was established using the previously described [1] CC10-tTS-rtTA-TGFβ1 transgenic (tgTGF-β1) mouse. The model was first assessed by investigating PCLS overexpression of TGF-β1 in response to stimulation of the transgene by doxycycline treatment. Gene expression of COX-2 and fibrotic markers including collagen were assessed after 4 days of treatment. The anti-fibrotic potential of 4 epigenetic inhibitors; BIX01294 (BIX, inhibitor of G9a histone methyltransferase), 3-deazaneplanocin A (DZNep, inhibitor of EZH2 histone methyltransferase), SAHA (inhibitor of histone deacetylases, HDACs) and Decitabine (DAC, DNA demethylating agent) was investigated. Viability of PCLS was assessed by MTT and Prestoblue® viability assay.
Results: Treatment of PCLS from tgTGF-β1 mice with doxycycline induced a concentration-dependent increase in global TGF-β1, pro-fibrotic markers including collagen and pro-inflammatory COX-2, which was comparable to recombinant TGF-β1 treatment. Treatment with three of the epigenetic inhibitors BIX01294, DZNep and DAC did not reduce the pro-fibrotic response following doxycycline treatment. However SAHA demonstrated a significant suppressive effect on COX-2 and collagen expression, while not directly affecting TGF-β1 transgene expression.
Conclusions: The data suggests that SAHA has the potential to reduce fibrosis in a TGF-β1 driven model of pulmonary fibrosis. Further work is currently underway to assess the anti-fibrotic potential of this drug in tgTGF-β1 animals.

Citation

Brand, O. J., Pasini, A., Habgood, A., Knox, A. J., Jenkins, G., & Pang, L. Suberanilohydroxamic acid (SAHA) inhibits collagen deposition in a transforming growth factor β1-driven precision cut lung slice (PCLS) model of pulmonary fibrosis. Presented at British Thoracic Society Winter Meeting 2016

• Presentation Conference Type: Conference Paper (published)
• Conference Name: British Thoracic Society Winter Meeting 2016
• End Date: Dec 9, 2016
• Acceptance Date: Oct 18, 2016
• Online Publication Date: Nov 15, 2016
• Publication Date: Dec 30, 2016
• Deposit Date: Nov 16, 2016
• Publicly Available Date: Nov 16, 2016
• Print ISSN: 0040-6376
• Electronic ISSN: 1468-3296
• Publisher: BMJ Publishing Group
• Peer Reviewed: Not Peer Reviewed
• Volume: 71
• Issue: Suppl. 3
• Pages: A31
• DOI: https://doi.org/10.1136/thoraxjnl-2016-209333.58
• Public URL: https://nottingham-repository.worktribe.com/output/822870
• Publisher URL: https://thorax.bmj.com/content/thoraxjnl/71/Suppl_3/A31.2.full.pdf
• Additional Information: Conference abstract from British Thoracic Society Winter Meeting 2016.
• Contract Date: Nov 16, 2016