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
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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

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Figure 4

S100a9K26 lactylation boosts the transcription of neutrophil migration and promotes neutrophil recruitment following MI/R.

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S100a9K26 lactylation boosts the transcription of neutrophil migration a...
(A) Immunoblotting for S100a9K26la, lamin B1, and GAPDH in cellular fractions of BM neutrophils from WT and S100a9K26R mice on post-MI/R day 1 (n = 4). (B and C) CUT-Tag analysis of S100a9K26la in blood neutrophils from sham-treated and post-MI/R mice on day 1. (B) Genes marked by exclusively increased in S100a9K26la (S100a9K26la-log2[MI/R/sham] ≥0.5 and S100a9-log2[MI/R/sham] ≤0.5, S100a9K26la-specific); increased in both S100a9K26la and S100a9 (S100a9K26la-log2[MI/R/sham] >1 and S100a9-log2[MI/R/sham] >1, shared); or exclusively increased in S100a9 (S100a9-log2[MI/R/sham] ≥0.5 and S100a9K26la-log2[MI/R/sham] ≤0.5, S100a9-specific). (C) Top 10 GO terms for genes with S100a9K26la-specific modifications. (D) Number of upregulated and downregulated genes with and without S100a9K26la binding. (E) GO terms of upregulated (red) and downregulated (blue) genes with S100a9K26la modification. (F) Representative migratory genes ranked by S100a9K26la binding signal (right) and mRNA expression according to the RNA-Seq data (left). (G and H) S100a9K26la occupancy (G) (n = 6) and gene expression (H) (n = 6) in circulating neutrophils at day 1 after MI/R were analyzed using ChIP-qPCR or RT-qPCR. (I) Adhesion of BM neutrophils on day 1 after MI/R in the presence or absence of Mg2+ (1 mmol/L) was determined on CMECs pretreated or untreated with TNF-α (20 ng/mL, n = 6). (J) Percentage of polarized neutrophils (with ruffled or extended pseudopods) on day 1 after MI/R after CXCL2 stimulation. Scale bars: 10 μm (n = 5). (K) Transwell assay of BM neutrophils on day 1 after MI/R with CXCL2 treatment (30 ng/mL) for 2 hours (n = 4). Median (IQR: 25th–75th percentiles). (L and M) Representative flow cytometry plots and quantification of BL (L) and heart (M) neutrophils (CD45+CD11b+Ly6G+) from WT and S100a9K26R mice on day 1 after MI/R (n = 5). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 for the indicated comparisons, by 1-way ANOVA with Tukey’s multiple-comparison test (P values were adjusted for 6 comparisons) (G and H), 2-way ANOVA with Tukey’s multiple-comparison test (P values were adjusted for 6 comparisons) (I), 2-tailed Mann-Whitney U test (K), and unpaired, 2-tailed Student’s t test (J, L, and M). All data indicate the mean ± SD.