Abstract
Subcellular machinery of NLRP3 is essential for inflammasome assembly and activation. However, the stepwise process and mechanistic basis of NLRP3 engagement with organelles remain unclear. Herein, we demonstrated glycogen synthase kinase 3β (GSK3β) as a molecular determinant for the spatiotemporal dynamics of NLRP3 inflammasome activation. Using live cell multispectral time-lapse tracking acquisition, we observed that upon stimuli NLRP3 was transiently associated with mitochondria and subsequently recruited to the Golgi network (TGN) where it was retained for inflammasome assembly. This occurred in relation to the temporal contact of mitochondria to Golgi apparatus. NLRP3 stimuli initiate GSK3β activation with subsequent binding to NLRP3, facilitating NLRP3 recruitment to mitochondria and transition to TGN. GSK3β activation also phosphorylates phosphatidylinositol 4-kinase 2 Α (PI4k2A) in TGN to promote sustained NLRP3 oligomerization. Our study has identified the interplay between GSK3β signaling and the organelles dynamics of NLRP3 required for inflammasome activation and opens new avenues for therapeutic intervention.
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Data availability
All data are presented in the main text or supplementary materials. The expression plasmids reported in this manuscript are available at Addgene or upon request. The rest of the data supporting the present study are available from the corresponding author upon reasonable request.
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Acknowledgements
We thank Yale Liver Center Morphology Core/Molecular Core Facilities for the support of confocal microcopy, image processing and biological assay, Yale Proteomics Core Facility for the support of LC-MS/MS assay, Center for cellular and molecular imaging (CCMI) EM core facility at Yale medical school for EM processing. This study was supported by NIH UO1 grant 5U01AA026962-04 (to WZM and XO), by VA Merit award BX003259-05(to W.Z.M.), by Yale Liver Center Award NIH P30DK-034989 Morphology Core and Cellular /Molecular Core, and in part by Yale Liver Center Pilot Project Award (to XO) and NCI grant 5R01CA224023-03 (to RAF). The Yale Liver Center Core Facilities were funded by NIH by grant DK P30-034989.
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SA, RAF, WZM, and XO designed the research plan and interpreted the results. SA, YQ, ZL, SH, QL, and XO performed and analyzed the experiments. SA and. SB performed reversal experiments. SA, WZM, and XO wrote and WZM and XO edited the manuscript. XO supervised the project and gave final approval.
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Our studies did not include human participants, human data, or human tissue. For the animal studies, experiments were performed according to both the Animal Care and Use Committee of Yale University and the Institutional Animal Ethics Committee/Institutional Animal Care and Use Committee (IACUC) of Army Medical University.
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Arumugam, S., Qin, Y., Liang, Z. et al. GSK3β mediates the spatiotemporal dynamics of NLRP3 inflammasome activation. Cell Death Differ 29, 2060–2069 (2022). https://doi.org/10.1038/s41418-022-00997-y
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DOI: https://doi.org/10.1038/s41418-022-00997-y
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