ZNF263-mediated transcriptional activation of HK3 alleviates sepsis-associated acute lung injury by regulating the PI3K/AKT/NF-κB pathway

Abstract Acute lung injury is a frequent complication of sepsis. Sepsis-associated acute lung injury (SALI) is a critical condition characterized by systemic inflammatory response syndrome, and effective targeted therapies remain limited. Hexokinase 3 (HK3), a glycolytic enzyme, has been implicated in immune regulation. However, the molecular role of HK3 in SALI remains unclear. This study aimed to investigate the regulatory mechanisms involving HK3 in the pathogenesis of SALI. Both in vivo and in vitro models of SALI were established to evaluate the effects of HK3 overexpression on inflammatory responses, apoptosis, and oxidative stress. Transcriptome sequencing was performed on HK3-overexpressing cells and corresponding controls to identify downstream pathways. Potential upstream transcription factors of HK3 were predicted through bioinformatic analysis, followed by rescue experiments to validate the regulatory axis. HK3 overexpression significantly reduced inflammatory factors, suppressed apoptosis, and alleviated oxidative stress in lung epithelial cells. In the SALI mouse model, HK3 overexpression alleviated lung pathology, reduced inflammatory cytokines in bronchoalveolar lavage fluid, decreased the number of apoptotic cells, and modulated oxidative stress markers. Transcriptomic sequencing and experimental results revealed that HK3 was associated with the inhibition of key components of the TNF/NF-κB signaling cascade, particularly the PI3K/AKT/NF-κB pathway. Zinc finger protein 263 (ZNF263) is a transcription factor that directly binds to the HK3 promoter and activates its transcription. ZNF263 overexpression upregulated HK3, attenuating LPS-induced inflammation, apoptosis, and oxidative damage. Rescue experiments further confirmed that ZNF263 exerts a lung-protective effect through HK3-associated modulation of the PI3K/AKT/NF-κB axis. This study reveals for the first time that the transcription factor ZNF263 mitigates inflammation, apoptosis, and oxidative stress in pulmonary epithelial cells during SALI by targeting HK3, a process that is associated with suppression of the PI3K/AKT/NF-κB pathway. Our findings identify a promising therapeutic target, providing a theoretical basis for developing novel treatment strategies for SALI.