Single‐Cell Reveal GALNT7‐Dependent Ferroptosis Suppression as a Mechanism of Immunotherapy Resistance in Non‐Small Cell Lung Cancer

ABSTRACT Immunotherapy has transformed the treatment of non‐small cell lung cancer (NSCLC), yet most patients fail to respond due to poorly understood resistance mechanisms. Here, integrative multi‐omics analysis combining single‐cell RNA‐sequencing, bulk RNA‐seq, and spatial transcriptomics is performed on tumors from NSCLC patients receiving immune checkpoint blockade (ICB). Transcriptomic profiling revealed that GALNT7, a glycosyltransferase, is selectively upregulated in non‐responders (NR) and enriched in malignant epithelial cells. Functional and pathway analyses linked GALNT7 expression to suppression of ferroptosis‐related signaling, whereas ICB responders (R) exhibited higher ferroptosis activity. Silencing GALNT7 in NSCLC cells impaired proliferation, induced apoptosis, and triggered ferroptotic cell death, characterized by lipid peroxidation and mitochondrial damage. Mechanistically, GALNT7 loss decreased SLC7A11 and GPX4, while upregulating the ferroptosis activator ACSL4. In vivo, GALNT7 knockdown reduced tumor growth, enhanced CD8 ⁺ T cell infiltration, and increased interferon gamma (IFN‐γ) production within the tumor microenvironment. Moreover, combining GALNT7 depletion with PD‐1 blockade achieved synergistic tumor suppression, which is reversed by Ferrostatin‐1, indicating ferroptosis‐dependent immunostimulation. Collectively, these findings uncover GALNT7 as a critical regulator linking ferroptosis and anti‐tumor immunity, providing a mechanistic basis for ferroptosis‐based sensitization to ICB therapy in NSCLC.