Jia‐Shen Wu, Xinyu Zhang, Xinyu Ma, Yue‐Yue Wei, Leilei Wang, Ze‐Bang Du, Xiaogang Xia, Lin Che, Dong‐Bei Guo, Hanying Zheng, Youliang Yao, Wengang Li, Yu‐Chun Lin, Zhong‐Ning Lin
Abstract Microcystin‐LR (MC‐LR) exacerbates metabolic dysfunction‐associated steatotic liver disease (MASLD) by inducing histopathological damage and lipid metabolism disorders. Inducible hepatocyte‐derived extracellular vesicles (iHD‐EVs) released after xenobiotic exposure activate the macrophage NOD‐like receptor protein 3 (NLRP3) inflammasome. Suppression of NLRP3 phosphorylation at serine 295 (p‐NLRP3 S295 ) is previously shown to alleviate MASLD progression. Here, it is demonstrated that microcystin‐LR (MC‐LR)‐induced iHD‐EVs reduced deliver of miR‐328‐3p to macrophages, thereby upregulating protein phosphatase 2A (PP2A)‐B56δ. Consequent PP2A‐B56δ activation disrupts inositol 1,4,5‐triphosphate receptor and voltage‐dependent anion channel 1 coupling, evokes mitochondria‐associated endoplasmic reticulum membrane (MAM) calcium (Ca 2+ ) overload, and recruits p‐NLRP3 S295 into the inflammasome. Neutralization of p‐NLRP3 S295 with a site‐specific monoclonal antibody (anti‐p‐NLRP3 S295 mAb) markedly attenuates liver inflammation and injury in MC‐LR‐exposed mice. Collectively, the miR‐328‐3p/PP2A‐B56δ/p‐NLRP3 S295 axis is identified as a crucial driver of metaflammation and establishes circulating EV‐miR‐328‐3p as a novel biomarker and anti‐p‐NLRP3 S295 mAbs as translational tools for MC‐LR‐associated MASLD.