Premium
Inhibition of Tumor Necrosis Factor Receptor 1 Signaling by Small Molecules
Author(s) -
Lo Chih Hung,
Vunnam Nagamani,
Lewis Andrew,
Chiu TingLan,
Brummel Benjamin,
Schaaf Tory,
Grant Benjamin,
Bawaskar Prachi,
Thomas David,
Sachs Jonathan
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.609.11
Subject(s) - förster resonance energy transfer , tumor necrosis factor alpha , receptor , tumor necrosis factor receptor 1 , microbiology and biotechnology , small molecule , signal transduction , extracellular , drug discovery , chemistry , ligand (biochemistry) , biophysics , etanercept , pharmacology , biochemistry , biology , immunology , fluorescence , tumor necrosis factor receptor , physics , quantum mechanics
Tumor necrosis factor receptor 1 (TNFR1) is a member of the TNFR superfamily which can be activated by binding to one of two cognate ligands, tumor necrosis factor (TNF‐α) or lymphotoxin‐α (LT‐α), via its extracellular domain. Ligand binding leads to network formation and signaling cascade of IκBα degradation and NF‐κB activation, resulting in inflammatory and autoimmune diseases. Current treatments, such as etanercept and infliximab, reduce symptoms through sequestration of free ligands. However, these treatments are expensive and induce dangerous side‐effects due to off‐target inhibition of other related TNF receptors that are not involved in the disease condition. As a consequence, there is a desperate need for receptor‐specific treatments, which hold the promise of overcoming the limitations of conventional therapeutics. It has been proposed that TNFR1 pre‐ligand assembly domain (PLAD), a portion of the extracellular region of TNFR1 that mediates receptor‐chain association essential for signaling, is an important therapeutic target in inflammatory arthritis. This motivates the discovery of small‐molecule inhibitors that could bind to TNFR1 PLAD to disrupt receptor‐receptor interactions and inhibit downstream signaling. In this study, we have engineered a biosensor exhibiting fluorescence resonance energy transfer (FRET) by attaching fluorophores to the intracellular domain of TNFR1. By using a fluorescence lifetime screening platform, two hit compounds were identified to inhibit TNFR1 function. These small‐molecule inhibitors reduce FRET and disrupt PLAD‐PLAD interaction without ablating ligand binding. The biosensor and the fluorescence lifetime screening approach reported here provide a very promising high‐throughput drug discovery platform for TNF receptors, as well as oligomeric receptors in general. Support or Funding Information This work is supported by National Institutes of Health Grant R01 GM107175 and R42 DA037622