RT Journal Article
T1 Methotrexate limits inflammation through an A20-dependent cross-tolerance mechanism
A1 Municio, Cristina
A1 Dominguez-Soto, Ángeles
A1 Fuentelsaz-Romero, Sara
A1 Lamana Domínguez, Amalia
A1 Montes, Nuria
A1 Cuevas, Víctor
A1 García Campos. Raquel, 
A1 Pablos Álvarez, José Luis
A1 González-Álvaro, Isidoro
A1 Puig-Kröger, Amaya
AB OBJECTIVES: Methotrexate (MTX) is the anchor drug for treatment of rheumatoid arthritis (RA), but the mechanism of its anti-inflammatory action is not fully understood. In RA, macrophages display a proinflammatory polarisation profile that resembles granulocyte-macrophage colony-stimulating factor (GM-CSF)-differentiated macrophages and the response to MTX is only observed in thymidylate synthase+ GM-CSF-dependent macrophages. To determine the molecular basis for the MTX anti-inflammatory action, we explored toll-like receptor (TLR), RA synovial fluid (RASF) and tumour necrosis factor receptor (TNFR)-initiated signalling in MTX-exposed GM-CSF-primed macrophages.METHODS:Intracellular responses to TLR ligands, TNFα or RASF stimulation in long-term low-dose MTX-exposed human macrophages were determined through quantitative real-time PCR, western blot, ELISA and siRNA-mediated knockdown approaches. The role of MTX in vivo was assessed in patients with arthritis under MTX monotherapy and in a murine sepsis model.RESULTS:MTX conditioned macrophages towards a tolerant state, diminishing interleukin (IL)-6 and IL-1β production in LPS, LTA, TNFα or RASF-challenged macrophages. MTX attenuated LPS-induced MAPK and NF-κB activation, and toll/IL-1R domain-containing adaptor inducing IFN-beta (TRIF1)-dependent signalling. Conversely, MTX increased the expression of the NF-κB suppressor A20 (TNFAIP3), itself a RA-susceptibility gene. Mechanistically, MTX-induced macrophage tolerance was dependent on A20, as siRNA-mediated knockdown of A20 reversed the MTX-induced reduction of IL-6 expression. In vivo, TNFAIP3 expression was significantly higher in peripheral blood cells of MTX-responsive individuals from a cohort of patients with arthritis under MTX monotherapy, whereas MTX-treated mice exhibited reduced inflammatory responses to LPS.CONCLUSIONS:MTX impairs macrophage proinflammatory responses through upregulation of A20 expression. The A20-mediated MTX-induced innate tolerance might limit inflammation in the RA synovial context, and positions A20 as a potential MTX-response biomarker.
PB BMJ Publishing Group
SN 0003-4967
YR 2018
FD 2018
LK https://hdl.handle.net/20.500.14352/94316
UL https://hdl.handle.net/20.500.14352/94316
LA eng
NO Municio C, Dominguez-Soto à , Fuentelsaz-Romero S, et alMethotrexate limits inflammation through an A20-dependent cross-tolerance mechanismAnnals of the Rheumatic Diseases 2018;77:752-759.
NO This work was supported by grants from Instituto de Salud Carlos III/FEDER (PI14/00075 and PI17/00037) to AP-K, PI14/00422 to IG-A, RIER RD16 to JLP, IG-A and AP-K, Ministerio de Economía y Competitividad SAF2014-54423-R to ALC. FEDER, Fondo Europeo de Desarrollo Regional: una manera de hacer Europa. SF-R and AP-K are supported by FIBHGM.
NO Instituto de Salud Carlos III
NO European Commission
NO Ministerio de Economía y Competitividad (España)
DS Docta Complutense
RD 9 abr 2025