Extracellular matrix stiffness orchestrates NETs formation and mTOR-Driven metabolic reprogramming in lung cancer

Lung cancer is strongly associated with increased extracellular matrix (ECM) stiffness, which correlates with poor patient prognosis. Our study reveals that high-stiffness tumor niches exhibit significant upregulation of neutrophil extracellular traps (NETs), which enhance the Warburg effect and promote tumor cell proliferation. Using atomic force microscopy (AFM) and multi-immunofluorescence staining (mIF), we demonstrated a spatial correlation between NETs formation and localized ECM stiffness in lung cancer tissues. In Kras^LSL-G12D/+/Trp53 ^fl/fl transgenic mouse model, bleomycin-induced lung stiffening further increased NETs generation, while genetic ablation of Pad4 (Pad4^-/- mouse model) or pharmacological inhibition of NETs (via LOX mAb, BAPN, or DNase I) reduced tumor burden. Mechanistically, stiffness-driven NETs upregulated glycolytic enzymes and extracellular acidification rate (ECAR) through aberrant mTOR pathway activation. Ex vivo and patient-derived xenograft (PDX) models validated the therapeutic potential of targeting LOX could extracellularly attenuate stiffness of ECM and intracellularly inhibit mTOR pathway. Our findings propose a novel strategy to improve lung cancer outcomes by disrupting the stiffness-NETs-mTOR axis, offering a one target-dual function approach for tumors with stiffening ECM.