Background Although precision-targeted therapies and tyrosine kinase inhibitors (TKIs) have significantly improved outcomes in non-small-cell lung cancer (NSCLC), patients with EGFR-mutant NSCLC with concurrent TP53 mutations often develop drug resistance and experience poor clinical outcomes. This study aims to investigate the molecular mechanisms underlying this aggressive subtype using single-cell RNA sequencing. Methods Formalin-fixed paraffin-embedded (FFPE) tumor samples were obtained from 40 hospitalized NSCLC patients. Somatic mutation profiles were determined using a targeted 23-gene next-generation sequencing (NGS) panel. Four samples harboring concurrent EGFR and TP53 mutations were selected for single-cell transcriptomic profiling using the 10x Genomics platform. Results Two dominant malignant epithelial cell populations were identified: C1EGFR +, associated with proliferation and invasion, and C2STAT1 +, linked to immunosuppression and drug resistance. These tumor subtypes cooperatively drive CD8 + T cell exhaustion through the MDK– (ITGA4+ITGB1), MIF– (CD74+CXCR4), and TGF-β signaling pathways. In addition, antigen-presenting cancer-associated fibroblasts (apCAFs) recruit regulatory T cells via the CCL5–CCR4 axis, collectively establishing an immune-excluded tumor microenvironment. Mechanistically, a STAT1/ETS1-centered transcriptional program regulates the expression of key immunosuppressive (e. g. , MDK, MIF, TGFB1) and resistance-associated genes (e. g. , ERBB2, JAK2). Conclusion These findings reveal a coordinated transcriptional network that promotes immune evasion and therapeutic resistance in EGFR/TP53 co-mutated NSCLC. Targeting the STAT1/ETS1 axis, in combination with EGFR-TKIs or immune checkpoint inhibitors, may provide a novel strategy to overcome resistance and improve patient outcomes. Further validation in larger patient cohorts and functional studies is warranted to confirm these observations and support clinical translation.
Shi et al. (Fri,) studied this question.