ALDOA maintains NLRP3 inflammasome activation by controlling AMPK activation

Dysregulated NLRP3 inflammasome activity results in uncontrolled inflammation, which is the basis of many chronic diseases. Although the regulatory mechanism has been gradually clarified after a long period of research, the metabolic regulation of NLRP3 inflammasome is still a mystery. Here, we find that ALDOA, as a monitor of glycolysis, regulates NLRP3 inflammasome by sensing changes in glycolytic flux to participate in the formation of AXIN-based AMPK-activation complex on the lysosomal surface. In this process, ALDOA restricts PRKN/parkin-dependent mitophagy through controlling AMPK activation to maintain mitochondrial damage caused by NLRP3 agonists. Furthermore, ALDOA also regulates the transcription of <i>SQSTM1/p62</i>, a receptor for mitophagy, through AMPK-FOXO3 signaling. In addition to studying the mechanism by which ALDOA regulated NLRP3 inflammasome, we also screened ALDOA inhibitors and found that LYG-202, a synthetic flavonoid compound, inhibited ALDOA enzyme activity by occupying the position involved in Schiff base intermediate formation, thus preventing FBP from combining with ALDOA. In vitro, LYG-202 suppressed NLRP3 inflammasome via activating the AMPK-mitophagy signaling pathway. In vivo, LYG-202 attenuated sterile inflammation and fulminant hepatitis, and suppressed the activation of NLRP3 inflammasome activation. Therefore, our study demonstrated that the glycolytic enzyme ALDOA maintained NLRP3 inflammasome activation by monitoring the glycolytic flux to control AMPK activation during the classical activation of NLRP3. ALDOA: aldolase A; AMPK: AMP-activated protein kinase; ATG: autophagy related; ATG5: autophagy related 5; ATP: adenosine triphosphate; BMDMs: bone marrow-derived macrophages; CALCOCO2: calcium binding and coiled-coil domain 2; CASP1: caspase 1; CQ: chloroquine; FOXO3: forkhead box O3; IL1B: interleukin 1 beta; LPS: lipopolysaccharide; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3 beta; MT: mutant; mtDNA: mitochondrial DNA; MTORC1: mechanistic target of rapamycin kinase complex 1; mtROS: mitochondrial reactive oxygen species; NLRP3: NLR family, pyrin domain containing 3; OPTN: optineurin; PBS: phosphate-buffered saline; PRKN/Parkin: parkin RBR E3 ubiquitin protein ligase; SN: supernatant; SQSTM1/p62: sequestosome 1; STK11/LKB1: serine/threonine kinase 11; TOMM20: translocase of outer mitochondrial membrane 20; ULK1: unc-51 like autophagy activating kinase 1; v-ATPase: vacuolar type H+-ATPase; WT: wild-type.