The transcriptomic landscape of canonical activation of NLRP3 inflammasome from bone marrow-derived macrophages

Abstract

The NLRP3 inflammasome has gained significant attention due to its participation in diverse cellular processes. Nevertheless, the detailed framework of the canonical NLRP3 inflammasome assembly still remains unrevealed. This study aims to elucidate the transcriptomic landscape of the various stages involved in the canonical activation of the NLRP3 inflammasome in BMDMs by integrating RNA-seq, bioinformatics, and molecular dynamics analyses. The model for the canonical activation of the NLRP3 inflammasome was confirmed through morphological observations, functional assessments (ELISA and LDH), and protein detection (western blot). Subsequently, cells were subjected to RNA sequencing following three groups: control, priming (LPS 500 ng/ml, 4 h), and activation (LPS 500 ng/ml, 4 h; ATP 5 mM, 1 h). A total of 9116 differentially expressed genes (DEGs) were identified, which exerted regulatory effects on various pathways, including cell metabolism, ion fluxes, post-translational modifications, and organelles. Subsequently, six hub genes (Sirt3, Stat3, Syk, Trpm2, Tspo, and Txnip) were identified via integrating literature review and database screening. Finally, the three-dimensional structures of these six hub proteins were obtained using the MD-optimized RoseTTAFold and Gromacs simulations (at least 200 ns). In summary, our research offers novel insights into the transcriptomic-level understanding of the assembly of the canonical NLRP3 inflammasome.