Effects of a novel microtubule-depolymerizer on pro-inflammatory signaling in RAW264.7 macrophages

The Nuclear Factor-kappa B (NF-kappa B) pathway is vital for immune system regulation and pro-inflammatory signaling. Many inflammatory disorders and diseases, including cancer, are linked to dysregulation of NF-kappa B signaling. When macrophages recognize the presence of a pathogen, the signaling pathway is activated, resulting in the nuclear translocation of the transcription factor, NF-kappa B, to turn on pro-inflammatory genes. Here, we demonstrate the effects of a novel microtubule depolymerizer, NT-07-16, a polysubstituted pyrrole compound, on this process. Treatment with NT-07-16 decreased the production of pro-inflammatory cytokines in RAW264.7 mouse macrophages. It appears that the reduction in pro-inflammatory mediators produced by the macrophages after exposure to NT-07-16 may be due to activities upstream of the translocation of NF-kappa B into the nucleus. NF-kappa B translocation occurs after its inhibitory protein, I kappa B-alpha is phosphorylated which signals for its degradation releasing NF-kappa B so it is free to move into the nucleus. Previous studies from other laboratories indicate that these processes are associated with the microtubule network. Our results show that exposure to the microtubule-depolymerizer, NT-07-16 reduces the phosphorylation of I kappa B-alpha and also decreases the association of NF-kappa B with tubulin which may affect the ability of NF-kappa B to translocate into the nucleus. Therefore, the anti-inflammatory activity of NT-07-16 may be explained, at least in part, by alterations in these steps in the NF-kappa B signaling pathway leading to less NF-kappa B entering the nucleus and reducing the production of pro-inflammatory mediators by the activated macrophages.