@article{WeineltKarathanasisSmithetal.2021,
  author    = {Weinelt, Nadine and Karathanasis, Christos and Smith, Sonja and Medler, Juliane and Malkusch, Sebastian and Fulda, Simone and Wajant, Harald and Heilemann, Mike and van Wijk, Sjoerd J. L.},
  title     = {Quantitative single-molecule imaging of TNFR1 reveals zafirlukast as antagonist of TNFR1 clustering and TNFα-induced NF-ĸB signaling},
  series = {Journal of Leukocyte Biology},
  volume    = {109},
  journal   = {Journal of Leukocyte Biology},
  number    = {2},
  doi       = {10.1002/JLB.2AB0420-572RR},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-215960},
  pages     = {363 -- 371},
  year      = {2021},
  abstract  = {TNFR1 is a crucial regulator of NF-ĸB-mediated proinflammatory cell survival responses and programmed cell death (PCD). Deregulation of TNFα- and TNFR1-controlled NF-ĸB signaling underlies major diseases, like cancer, inflammation, and autoimmune diseases. Therefore, although being routinely used, antagonists of TNFα might also affect TNFR2-mediated processes, so that alternative approaches to directly antagonize TNFR1 are beneficial. Here, we apply quantitative single-molecule localization microscopy (SMLM) of TNFR1 in physiologic cellular settings to validate and characterize TNFR1 inhibitory substances, exemplified by the recently described TNFR1 antagonist zafirlukast. Treatment of TNFR1-mEos2 reconstituted TNFR1/2 knockout mouse embryonic fibroblasts (MEFs) with zafirlukast inhibited both ligand-independent preligand assembly domain (PLAD)-mediated TNFR1 dimerization as well as TNFα-induced TNFR1 oligomerization. In addition, zafirlukast-mediated inhibition of TNFR1 clustering was accompanied by deregulation of acute and prolonged NF-ĸB signaling in reconstituted TNFR1-mEos2 MEFs and human cervical carcinoma cells. These findings reveal the necessity of PLAD-mediated, ligand-independent TNFR1 dimerization for NF-ĸB activation, highlight the PLAD as central regulator of TNFα-induced TNFR1 oligomerization, and demonstrate that TNFR1-mEos2 MEFs can be used to investigate TNFR1-antagonizing compounds employing single-molecule quantification and functional NF-ĸB assays at physiologic conditions.},
  language  = {en}
}