Multi-omics Analyses Identify ANLN as a Prognostic Biomarker for Recurrence and Metastasis in Non–Small Cell Lung Cancer

Abstract

Lung cancer recurrence and metastasis remain major causes of cancer-related mortality, yet the molecular determinants driving these processes are not fully defined. Herein we did integrated analyses of bulk transcriptomic data and single-cell RNA sequencing data from multi-cohorts and identified key regulators of lung cancer progression. By integrating gene features related to survival, recurrence, and metastasis from independent cohorts and further analyzing the composition of the tumor microenvironment, we identified ANLN as a core progression-related gene with poor prognosis. Its expression was elevated in recurrent and metastatic tumors and correlated with reduced overall survival. xCell analysis revealed epithelial enrichment and relative immune and stromal depletion in ANLN-high tumors. Single-cell analyses of lymph node metastases demonstrated that ANLN is predominantly expressed in epithelial and proliferating tumor cells and is associated with extensive transcriptional and remodeling of the tumor microenvironment. Functional scoring and enrichment analyses revealed that ANLN-high epithelial cells exhibit coordinated activation of proliferative and migratory programs alongside suppression of immune-associated features. Pseudotime trajectory analysis further positioned ANLN enrichment at a critical intermediate state during AT2-derived epithelial evolution toward an invasive phenotype. These findings were further validated in an independent single-cell dataset capturing the transition from in situ to invasive lung cancer. In summary, our results identify ANLN as a marker of a conserved, invasion-prone epithelial state underlying lung cancer recurrence and metastasis, providing mechanistic insights and potential therapeutic implications.