PLK1 inhibition dampens NLRP3 inflammasome–elicited response in inflammatory disease models
Marta Baldrighi, … , Ziad Mallat, Xuan Li
Marta Baldrighi, … , Ziad Mallat, Xuan Li
Published September 12, 2023
Citation Information: J Clin Invest. 2023;133(21):e162129. https://doi.org/10.1172/JCI162129.
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Research Article Cell biology Immunology

PLK1 inhibition dampens NLRP3 inflammasome–elicited response in inflammatory disease models

Abstract

Unabated activation of the NLR family pyrin domain–containing 3 (NLRP3) inflammasome is linked with the pathogenesis of various inflammatory disorders. Polo-like kinase 1 (PLK1) has been widely studied for its role in mitosis. Here, using both pharmacological and genetic approaches, we demonstrate that PLK1 promoted NLRP3 inflammasome activation at cell interphase. Using an unbiased proximity-dependent biotin identification (Bio-ID) screen for the PLK1 interactome in macrophages, we show an enhanced proximal association of NLRP3 with PLK1 upon NLRP3 inflammasome activation. We further confirmed the interaction between PLK1 and NLRP3 and identified the interacting domains. Mechanistically, we show that PLK1 orchestrated the microtubule-organizing center (MTOC) structure and NLRP3 subcellular positioning upon inflammasome activation. Treatment with a selective PLK1 kinase inhibitor suppressed IL-1β production in in vivo inflammatory models, including LPS-induced endotoxemia and monosodium urate–induced peritonitis in mice. Our results uncover a role of PLK1 in regulating NLRP3 inflammasome activation during interphase and identify pharmacological inhibition of PLK1 as a potential therapeutic strategy for inflammatory diseases with excessive NLRP3 inflammasome activation.

Authors

Marta Baldrighi, Christian Doreth, Yang Li, Xiaohui Zhao, Emily Warner, Hannah Chenoweth, Kamal Kishore, Yagnesh Umrania, David-Paul Minde, Sarah Thome, Xian Yu, Yuning Lu, Alice Knapton, James Harrison, Murray Clarke, Eicke Latz, Guillermo de Cárcer, Marcos Malumbres, Bernhard Ryffel, Clare Bryant, Jinping Liu, Kathryn S. Lilley, Ziad Mallat, Xuan Li

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Figure 5

PLK1 regulates microtubule nucleation and affects NLRP3 inflammasome positioning.

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PLK1 regulates microtubule nucleation and affects NLRP3 inflammasome pos...
(A and B) RosaCreErt2/WT Plk1fl/fl BMDMs were treated with 4-OH Tamoxifen (0.002 mg/mL 4-OH Tam, 24 hours) before and during priming (100 ng/mL LPS, 5 hours) and were then activated (5 mM ATP, 30 minutes). γ-Tubulin fluorescence was quantified within a 3 × 3 μm ROI centered around the main γ-tubulin focus. Scale bars: 10 μm (lower magnification) and 3 μm (enlarged insets) (A). Quantification by fluorescence intensity (n = 20) (B). (C and D) RosaCreErt2/WT Plk1fl/fl BMDMs were treated as in A. EB1 fluorescence was quantified in a circular ROI of 3 μM diameter. Scale bars: 3 μm (C). Quantification by mean EB1 fluorescence (No Tam, n = 54; 4-OH Tam, n = 77) (D). (E) RosaCreErt2/WT Plk1fl/fl BMDMs were treated with 4-OH tamoxifen (0.002 mg/mL 4-OH Tam, 24 hours) before and during priming (100 ng/mL LPS, 5 hours). Nlrp3 relative expression was quantified by qPCR. n = 3/group. (F) RosaCreErt2/WT Plk1fl/fl BMDMs were treated as in A and processed for fractionation Western blotting. cytop, cytoplasmic; memb, membrane; ins, insoluble fractions. GAPDH, GRASP65, and vimentin, respectively, were used as markers for each fraction. (F) Results are representative of 2 independent experiments. (A–F) Treatments without 4-OH tamoxifen were used as controls. Two-way ANOVA with Šidák’s post hoc test (B) and an unpaired t test (D and E) were used for statistical analysis. All data are the mean ± SEM.