2-Deoxy-D-glucose reshapes the sepsis-trained stressed granulocytes by comprehensive alterations of the transcriptional landscape

The secondary infections following sepsis pose a significant clinical challenge due to hyperinflammatory organ injury, which is primarily driven by maladaptive trained immunity (TRIM) in stressed granulocytes. Glucose metabolic reprogramming serves as a crucial mechanism underlying TRIM and is increasingly acknowledged as a promising therapeutic target for preventing cytokine storms following reinfections. Here, we employed recovery mice subsequent to the cecal ligation and puncture (CLP) procedures to model reinfection following sepsis and confirm the beneficial effects of 2-Deoxy-D-glucose (2-DG), which inhibits boosted glucose metabolism in stressed granulocytes. The 2-DG treatment leads to a marked reduction in TNF-α secretion, reactive oxygen species (ROS) production, and neutrophil extracellular traps (NETs) formation upon secondary stimulation while alleviating pulmonary inflammation in vivo. We conducted RNA sequencing to investigate how 2-DG treatment reshapes the transcriptomic profile of stressed granulocytes. Our findings revealed the effective reversion of LPS-induced pro-inflammatory responses after sepsis training, including cell migration, chemotaxis, and immune cell recruitment. However, 2-DG also caused extensive and non-specific suppressions of granulocyte biofunctions, such as cell-to-cell signaling and nucleic acid metabolism, indicating potential unexpected consequences. Furthermore, analysis of the upstream regulators in the 2DG-associated gene network identified CD44 as a promising alternative therapeutic target to 2-DG. This study provides novel insights into the pharmacological mechanism network associated with 2-DG treatment and supports the implementation of immunosuppressive strategies via targeted remodeling of glycose metabolism. This approach may serve as an effective therapeutic intervention for addressing secondary infections in sepsis.