S100a9 lactylation triggers neutrophil trafficking and cardiac inflammation in myocardial ischemia/reperfusion injury
Xiaoqi Wang, Xiangyu Yan, Ge Mang, Yujia Chen, Shuang Liu, Jiayu Sui, Zhonghua Tong, Penghe Wang, Jingxuan Cui, Qiannan Yang, Yafei Zhang, Dongni Wang, Ping Sun, Weijun Song, Zexi Jin, Ming Shi, Peng Zhao, Jia Yang, Mingyang Liu, Naixin Wang, Tao Chen, Yong Ji, Bo Yu, Maomao Zhang
Xiaoqi Wang, Xiangyu Yan, Ge Mang, Yujia Chen, Shuang Liu, Jiayu Sui, Zhonghua Tong, Penghe Wang, Jingxuan Cui, Qiannan Yang, Yafei Zhang, Dongni Wang, Ping Sun, Weijun Song, Zexi Jin, Ming Shi, Peng Zhao, Jia Yang, Mingyang Liu, Naixin Wang, Tao Chen, Yong Ji, Bo Yu, Maomao Zhang
View: Text | PDF
Research Article Cardiology Immunology

S100a9 lactylation triggers neutrophil trafficking and cardiac inflammation in myocardial ischemia/reperfusion injury

Abstract

Lactylation, a posttranslational modification derived from glycolysis, plays a pivotal role in ischemic heart disease. Neutrophils are predominantly glycolytic cells that trigger intensive inflammation of myocardial ischemia/reperfusion (MI/R). However, whether lactylation regulates neutrophil function during MI/R remains unknown. We applied lactyl proteomics analysis and found that S100a9 was lactylated at lysine 26 (S100a9K26la) in neutrophils, with elevated levels observed in both patients with acute myocardial infarction (AMI) and MI/R model mice. We demonstrated that S100a9K26la drove the development of MI/R using mutant knockin mice. Mechanistically, lactylated S100a9 translocated to the nucleus of neutrophils, where it bound to the promoters of migration-related genes, thereby enhancing their transcription as a coactivator and promoting neutrophil migration and cardiac recruitment. Additionally, lactylated S100a9 was released during neutrophil extracellular trap (NET) formation, leading to cardiomyocyte death by disrupting mitochondrial function. The enzyme dihydrolipoyllysine-residue acetyltransferase (DLAT) was identified as the lactyltransferase facilitating neutrophil S100a9K26la following MI/R, a process that could be restrained by α-lipoic acid. Consistently, we found that targeting the DLAT/S100a9K26la axis suppressed neutrophil burden and improved cardiac function following MI/R. In patients with AMI, elevated S100a9K26la levels in plasma were positively correlated with cardiac death. These findings highlight S100a9 lactylation as a potential therapeutic target for MI/R and as a promising biomarker for evaluating poor MI/R outcomes.

Authors

Xiaoqi Wang, Xiangyu Yan, Ge Mang, Yujia Chen, Shuang Liu, Jiayu Sui, Zhonghua Tong, Penghe Wang, Jingxuan Cui, Qiannan Yang, Yafei Zhang, Dongni Wang, Ping Sun, Weijun Song, Zexi Jin, Ming Shi, Peng Zhao, Jia Yang, Mingyang Liu, Naixin Wang, Tao Chen, Yong Ji, Bo Yu, Maomao Zhang

×

Figure 5

DLAT-catalyzed S100a9 lactylation in neutrophils can be antagonized by ALA.

Options: View larger image (or click on image) Download as PowerPoint
DLAT-catalyzed S100a9 lactylation in neutrophils can be antagonized by A...
(A) Differentially abundant proteins were identified in neutrophils by IP and MS analysis using anti-S100a9K26la and IgG antibodies. (B) MS/MS spectrum of DLAT. (C) IP assay of BM neutrophils following MI/R with the indicated antibodies (n = 4). (D) SPR analysis of lactyl-CoA binding to the recombinant DLAT protein. (E) An in vitro lactylation assay was performed in the presence or absence of lactyl-CoA (n = 4). (F) 32Dcl3 cells were differentiated into granulocytes with addition of G-CSF for 4 days and transfected with control siRNA (NC) or DLAT specific siRNA (siDLAT) for 24 hours, and then collected for immunoblot analysis (n = 4). (G) Molecular docking between DLAT and lactyl-CoA. (H) Per-residue energy contributions of key residues involved in lactyl-CoA combined with DLAT. (I) Purified rS100a9 was incubated with immunoprecipitates of WT Flag-DLAT or the I423A mutant from HEK293T cells followed by detection of S100a9K26 lactylation (n = 6). (J and K) BM neutrophils were stimulated with the indicated doses of ALA and then collected for IP with anti-DLAT (J) (n = 4) and intracellular acetyl-CoA detection by ELISA (K) (n = 6). Data indicate the mean ± SD. *P < 0.05 and **P < 0.01 for the indicated comparisons, by 1-way ANOVA with Tukey’s multiple-comparison test (K). (P values were adjusted for 6 comparisons.) (L) Purified rS100a9 was incubated with immunoprecipitates of WT Flag-DLAT or the K131/258R mutant (K2R)) from HEK293T cells followed by detection of S100a9K26la (n = 6). (M) In vitro lactylation assay in the presence or absence of acetyl-CoA (n = 4). (N and O) SPR analysis of acetyl-CoA (N) and ALA (O) binding to the recombinant DLAT protein. (P and Q) Flag DLAT plasmid was transfected into HEK293T cells, and then cell lysates were collected and incubated with vehicle or lactyl-CoA, acetyl-CoA, or ALA, followed by a CETSA (n = 4).