Microtubule affinity regulating kinase 4 promoted activation of the NLRP3 inflammasome-mediated pyroptosis in periodontitis

Abstract

Background Microtubule dynamics plays a crucial role in the spatial arrangement of cell organelles and activation of the NLRP3 inflammasome. Purpose This study aimed to explore whether microtubule affinity regulating kinase 4 (MARK4) can be a therapeutic target of periodontitis by affecting microtubule dynamics and NLRP3 inflammasome-mediated pyroptosis in macrophages. Materials and Methods The NLRP3 inflammasome-related genes and MARK4 were measured in the healthy and inflamed human gingival tissues. Bone marrow-derived macrophages (BMDMs) were infected with Porphyromonas gingivalis, while the MARK4 inhibitors (OTSSP167 and Compound 50) and small interference RNA were utilized to restrain MARK4. Apoptosis-associated speck-like protein (ASC) speck was detected by confocal, and levels of interleukin-1β (IL-1β), as well as IL-18, were assessed by ELISA. Results Increased staining and transcription of MARK4, NLRP3, ASC, and Caspase-1 were observed in the inflamed gingiva. P. gingivalis infection promoted MARK4 expression and the NLRP3 inflammasome in BMDMs. Inhibition of MARK4 decreased LDH release, IL-1β and IL-18 production, ASC speck formation, and the pyroptosis-related genes transcription. Furthermore, MARK4 inhibition reduced microtubule polymerization and acetylation in P. gingivalis- infected BMDMs. Conclusions MARK4 promoted NLRP3 inflammasome activation and pyroptosis in P. gingivalis- infected BMDMs by affecting microtubule dynamics. MARK4 inhibition might be a potential target in regulating the NLRP3 inflammasome during periodontitis progress.

Pyroptosis may play a crucial role in modifications of the immune microenvironment in periodontitis

“…Inflammasome activation is the basis of pyroptosis, and most studies reported the involvement of pyroptosis-related NLRP3 inflammasome in the pathogenesis of periodontitis. For instance, Wang et al 27 found that microtubule affinity regulating kinase 4 promoted NLRP3 inflammasome activation and pyroptosis in periodontitis; Xia et al 28 discovered miR-223-3p in salivary exosomes could modulate GSDMDmediated pyroptosis by targeting NLRP3 in periodontitis; Zhou et al 29 detected that pyroptosis may affect the onset and development of diabetes-associated periodontitis through activating NLRP3 inflammasome, which can also result in alveolar bone loss. 30 Up to date, research about pyroptosis is mainly focused on its influence in the tumor immune microenvironment.…”

Pyroptosis in bone loss

“…have found that porphyromonas gingivalis could activate innate immune cells through activating NLRP3 inflammasome, inducing proinflammatory cytokines during pyroptosis and subsequently enhancing alveolar bone loss [ 116 ]. Moreover, in the macrophages infected by porphyromonas gingivalis, overexpressed microtubule affinity regulating kinase 4 (MARK4) could further increased inflammasome activation and the pyroptosis, promoting alveolar bone loss [ 117 ]. Mycoplasma salivarium had the similar role with porphyromonas gingivali s , which could activate NLRP3 to induce IL-1β and promote periodontal disease accompanying bone loss [ 118 ].…”

“…Caspase-1 and caspase-4/-5/-11 cleave GSDMD into a GSDMD-N-terminal domain that anchors in the cell membrane, leading to the cell rupture. MARK4 [ 117 ], mycoplasma salivarium [ 118 ], staphylococcus aureus [ 119 ], and porphyromonas gingivalis [ 116 ] have been found in the presented pathway positively targeting NLRP3 to facilitate pyroptosis, yet glyburide [ 120 ], dioscin [ 91 ], metformin [ 121 ], resveratrol [ 122 ] target to suppress NLRP3. Meanwhile, caspase-1 is also the target of action to be modulated by P2X7/AMPK/cyclic stretch [ 124 , 125 ] and DEX [ 126 ], cranberry PACs [ 127 ], affecting bone loss involved in pyroptosis.…”

Pyroptosis in inflammatory bone diseases: Molecular insights and targeting strategies

“…Nrf2/HO-1 axis inhibits NF-κB pathway to attenuate pyroptosis 88 SDF-1 activates AMPK signal to suppress NLRP3 inflammasome and subsequent pyroptosis 89 A3AR inhibits the ROS/NLPR3/GSDMD axis to relieve pyroptosis-mediated symptoms in OA 90 Osteomyelitis NLRP3 inflammasome is activated to cause bone loss 93 Periodontitis Hypoxia with P. gingivalis-LPS synergistically activate NLRP3 to induce pyroptosis 101 MARK4 induces NLRP3-mediated pyroptosis 102 The Nrf2/HO-1 axis inhibits LPS-induced ROS to prevent NLRP3-mediated pyroptosis 98 NLRP6 inflammasome activates caspase-1 to induce pyroptosis of HGFs 103 Caspase-4/GSDMD pathway mediates noncanonical pyroptosis 104,105 Butyrate activates caspase-3/GSDME axis to induce pyroptosis 106 LPS-induced Dec1 upregulates NF-κB via the PI3K/AKT pathway to promote pyroptosis, while LPS-induced Dec2 attenuates the phosphorylation of NF-κB to inhibit pyroptosis [108][109][110][111] RA TNF-α plus CRT induces NLRP3 inflammasome-mediated pyroptosis 116 ACPAs induces NLRP3 inflammasome-mediated pyroptosis via the Akt/NF-κB axis 117 S100A9 may participate in the IL-6-induced pyroptosis via activating the CTSB/ATP pathway 118 Hypoxia induces NLRP3 inflammasome-mediated pyroptosis via the ROS/GRK2/HIF-1α axis 119 TNF-α plus hypoxia induce caspase-3/GSDME-mediated pyroptosis 120 ASIC1a promotes the influx of Ca 2+ , activating the calpain/calcineurin axis to induce NLRP3 inflammasome-mediated pyroptosis 122,123 CaSR promotes the influx of Ca 2 to induce NLRP3 inflammasome-mediated pyroptosis 124 Abbreviations: ACPAs, anti-citrullinated protein antibodies; AP, apical periodontitis; ASIC1a, acid-sensitive ion channel 1a; ATP, adenosine triphosphate; A3AR, adenosine A3 receptor; CaSR, calcium-sensing receptor; CRT, calreticulin; CTSB, cathepsin B; Dec1, embryo-chondrocyte expressed genes 1; FLS, fibroblast-like synoviocyte; Gout, gouty arthritis; GRK2, G protein-coupled receptor kinase 2; GSDMD, gasdermin D; HGFs, human gingival fibroblasts; HIF-1α, hypoxia-inducible factor-1α; HO-1, heme oxygenase-1; IL, interleukin; LPS, lipopolysaccharide; MARK4, microtubule affinity regulating kinase 4; NF-κB, nuclear factor kappa-B; NLRP3, nucleotide-binding oligomerization domain receptor family PYD domain containing 3; NOX, NAD(P)H oxidases; Nrf2, nuclear factor erythroid-2-related factor 2; OA, osteoarthritis; P. gingivalis, Porphyromonas gingivalis; ROS, reactive oxygen species; SDF-1, stromal cell-derived factor-1; TNF-α, tumor necrosis factor-α; USP7, ubiquitin-specific protease 7.…”

“…100,101 Thus, a combination of hypoxia and P. gingivalis-LPS synergistically activated the NLRP3 inflammasome in HGFs to promote pyroptosis in periodontitis. Recently, Wang et al 102 found microtubule affinity regulating kinase 4 (MARK4) could affect microtubule dynamics to activate NLRP3 inflammasome, illustrating that NLRP3 inflammasome-mediated pyroptosis may be regulated by inhibiting MARK4 during periodontitis progression. In contrast, activating the Nrf2/HO-1 axis could inhibit pyroptosis by decreasing the NLRP3 inflammasome and ROS caused by LPS, indicating that the Nrf2/HO-1 pathway is a periodontitisprotective mechanism.…”