BACKGROUND: Tumor-associated macrophages (TAMs) play pivotal roles in innate immunity and contribute to the advancement of lung cancer. We aimed to identify novel TAM-related biomarkers and significance of macrophage infiltration in lung adenocarcinoma (LUAD) through an integrative analysis of single-cell RNA-sequencing (scRNA-seq) data. To describe the cell atlas and construct a novel prognostic signature in LUAD. METHODS: The gene signature linked to TAMs was identified utilizing Scanpy from the scRNA-seq dataset GSE131907. Subsequent analysis involved evaluating the expression levels of these genes, their potential molecular mechanisms, and prognostic significance in LUAD using data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. We also constructed a risk score models through LASSO Cox regression for these genes. The underlying mechanism was further elucidated through the application of GSEA, ESTIMATE, TIDE, and other bioinformatic algorithms. RESULTS: Single-cell atlas was described by analyze 29 scRNA-seq samples from 19 LUAD patients. The TAMs-related gene signature (TGS) was identified as an independent prognostic factor by LASSO Cox regression analysis using differential expression genes (DEGs) derived from pro- and anti-inflammatory macrophage cells. Risk score model including nine TAMs-related genes (FOSL1, ZNF697, ADM, UBE2S, TICAM1, S100P, BIRC3, TLE1, and DEFB1) were obtained for prognosis construction. Moreover, the risk model underwent additional validation in four external GEO cohorts: GSE31210, GSE72094, GSE26939, and GSE30219. Interestingly, TGS-high tumors revealed enrichments in TGF-β signaling and hypoxia pathways, which shown low immune infiltration and immunosuppression by ESTIMATE and TIDE algorithm. The TGS-high risk group exhibited lower richness and diversity in the T-cell receptor (TCR) repertoire. CONCLUSION: This study introduces a novel TGS score developed through LASSO Cox regression analysis, utilizing DEGs in pro- and anti-inflammatory macrophage cells. High TGS tumors exhibited enrichment in TGF-β signaling and hypoxia pathways, suggesting their potential utility in predicting prognosis and immune responses in patients with LUAD. These results offer promising implications for the development of therapeutic strategies for LUAD.