A signalome screening approach in the autoinflammatory disease TNF receptor associated periodic syndrome (TRAPS) highlights the anti-inflammatory properties of drugs for repurposing. (November 2017)

• Record Type: Journal Article
• Authors: Todd, Ian; Negm, Ola H.; Reps, Jenna; Radford, Paul; Figueredo, Grazziela; McDermott, Elizabeth M.; Drewe, Elizabeth; Powell, Richard J.; Bainbridge, Susan; Hamed, Mohamed; Crouch, Sharon; Garibaldi, Jon; St-Gallay, Steve; Fairclough, Lucy C.; Tighe, Patrick J.
• Is Part Of: Pharmacological research. Volume 125(2017)Part B
• Page Start: 188
• Page End: 200
• Publication Date: 2017-11
• Subjects: BCA bicinchoninic acid -- CV coefficient of variation -- MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide -- PBMCs peripheral blood mononuclear cells -- RPPA reverse phase protein array -- SSMD strictly standardized mean difference -- TNFR1 tumour necrosis factor receptor-1 -- TRAPS TNF receptor associated periodic syndrome -- WT wild type; Drug repurposing -- Reverse-phase protein micro-array -- Signalome -- TNF receptor associated periodic syndrome -- Fluoroquinolone -- Anti-inflammatory; Pharmacology -- Periodicals; Pharmacology -- Periodicals; Research -- Periodicals; Médicaments -- Recherche -- Périodiques; Pharmacologie -- Périodiques; 615.105
• DOI: 10.1016/j.phrs.2017.08.012
• Languages: English
• ISSNs: 1043-6618

Abstract

Graphical abstract: Abstract: TNF receptor associated periodic syndrome (TRAPS) is an autoinflammatory disease caused by mutations in TNF Receptor 1 (TNFR1). Current therapies for TRAPS are limited and do not target the pro-inflammatory signalling pathways that are central to the disease mechanism. Our aim was to identify drugs for repurposing as anti-inflammatories based on their ability to down-regulate molecules associated with inflammatory signalling pathways that are activated in TRAPS. This was achieved using rigorously optimized, high through-put cell culture and reverse phase protein microarray systems to screen compounds for their effects on the TRAPS-associated inflammatory signalome. 1360 approved, publically available, pharmacologically active substances were investigated for their effects on 40 signalling molecules associated with pro-inflammatory signalling pathways that are constitutively upregulated in TRAPS. The drugs were screened at four 10-fold concentrations on cell lines expressing both wild-type (WT) TNFR1 and TRAPS-associated C33Y mutant TNFR1, or WT TNFR1 alone; signalling molecule levels were then determined in cell lysates by the reverse-phase protein microarray. A novel mathematical methodology was developed to rank the compounds for their ability to reduce the expression of signalling molecules in the C33Y-TNFR1 transfectants towards the level seen in the WT-TNFR1 transfectants. Seven high-ranking drugs were selected and tested by RPPA for effects on the same 40 signalling molecules in lysates of peripheral blood mononuclear cells (PBMCs) from C33Y-TRAPS patients compared to PBMCs from normal controls. The fluoroquinolone antibiotic lomefloxacin, as well as others from this class of compounds, showed the most significant effects on multiple pro-inflammatory signalling pathways that are constitutively activated in TRAPS; lomefloxacin dose-dependently significantly reduced expression of 7/40 signalling molecules across the Jak/Stat, MAPK, NF-κB and PI3K/AKT pathways. This study demonstrates the power of signalome screening for identifying candidates for drug repurposing.