title: Mechanisms of peroxisome proliferator activated receptor γ regulation by non-steroidal anti-inflammatory drugs.
abstract: Non-steroidal anti-inflammatory drugs (NSAIDs) display anti-inflammatory, antipyretic and analgesic properties by inhibiting cyclooxygenases and blocking prostaglandin production. Previous studies, however, suggested that some NSAIDs also modulate peroxisome proliferator activated receptors (PPARs), raising the possibility that such off target effects contribute to the spectrum of clinically relevant NSAID actions. In this study, we set out to understand how peroxisome proliferator activated receptor-γ (PPARγ/PPARG) interacts with NSAIDs using X-ray crystallography and to relate ligand binding modes to effects on receptor activity. We find that several NSAIDs (sulindac sulfide, diclofenac, indomethacin and ibuprofen) bind PPARγ and modulate PPARγ activity at pharmacologically relevant concentrations. Diclofenac acts as a partial agonist and binds to the PPARγ ligand binding pocket (LBP) in typical partial agonist mode, near the β-sheets and helix 3. By contrast, two copies of indomethacin and sulindac sulfide bind the LBP and, in aggregate, these ligands engage in LBP contacts that resemble agonists. Accordingly, both compounds, and ibuprofen, act as strong partial agonists. Assessment of NSAID activities in PPARγ-dependent 3T3-L1 cells reveals that NSAIDs display adipogenic activities and exclusively regulate PPARγ-dependent target genes in a manner that is consistent with their observed binding modes. Further, PPARγ knockdown eliminates indomethacin activities at selected endogenous genes, confirming receptor-dependence of observed effects. We propose that it is important to consider how individual NSAIDs interact with PPARγ to understand their activities, and that it will be interesting to determine whether high dose NSAID therapies result in PPAR activation.
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Ana C
authors.initials: AC
authors.lastname: Puhl
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Flora A
authors.initials: FA
authors.lastname: Milton
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Cvoro
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Douglas H
authors.initials: DH
authors.lastname: Sieglaff
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Jéssica C L
authors.initials: JC
authors.lastname: Campos
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Amanda
authors.initials: A
authors.lastname: Bernardes
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Carly S
authors.initials: CS
authors.lastname: Filgueira
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Jan Lammel
authors.initials: JL
authors.lastname: Lindemann
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Tuo
authors.initials: T
authors.lastname: Deng
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Francisco A R
authors.initials: FA
authors.lastname: Neves
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Igor
authors.initials: I
authors.lastname: Polikarpov
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador Saocarlense 400, São Carlos, SP, 13560-970, Brazil (ACP, JCLC, AB, IP).
authors.forename: Paul
authors.initials: P
authors.lastname: Webb
mesh_terms: D041721:3T3-L1 Cells; D050156:Adipogenesis / Q000187:drug effects; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D054333:Drug Partial Agonism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D047495:PPAR gamma / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D047495:PPAR gamma
keywords: 3T3-L1; X-ray structure; gene expression; non-steroidal anti-inflammatory drugs; partial agonist; peroxisome proliferator activated receptors