A kbhb-associated prognostic signature identifies TTLL12 as a promoter of lung adenocarcinoma progression associated with M2 macrophage infiltration

Lysine β-hydroxybutyrylation (kbhb) is a post-translational modification closely linked to cellular metabolic status and contributes to tumor initiation and progression. However, the prognostic value and immunological relevance of kbhb-associated genes in lung adenocarcinoma (LUAD) have not been systematically characterized. Kbhb-associated differentially expressed genes were identified from the TCGA and GEO datasets, and a prognostic model was constructed using machine learning algorithms. Differences in clinical outcomes, immune profiles, and drug sensitivity were then systematically compared across distinct risk groups. Furthermore, single-cell omics, virtual screening, molecular dynamics simulations, and in vitro experiments were integrated to comprehensively evaluate the clinical significance, biological functions, and potential therapeutic relevance of the core gene TTLL12. A 14-gene prognostic model based on the CoxBoost+Ridge algorithm showed stable predictive performance across multiple cohorts. The high-risk group was mainly enriched in cell cycle- and DNA replication-related pathways and exhibited immunosuppression-related features and distinct drug sensitivity profiles. The core gene TTLL12 was upregulated in LUAD and was associated with advanced clinical stage and poor prognosis. Single-cell analysis and in vitro experiments suggested that TTLL12 may be associated with enhanced malignant phenotypes in LUAD cells and M2 macrophage polarization. Virtual screening indicated that relatively stable binding was observed between Paryriogenin I and TTLL12, suggesting that Paryriogenin I may be a candidate compound for further evaluation. The kbhb-related prognostic model established in this study may provide a useful tool for risk stratification in LUAD. Notably, TTLL12 is a promising prognostic biomarker that promotes tumor progression, and this process may be potentially associated with M2 macrophage polarization. In addition, the identification of Paryriogenin I may provide a theoretical reference for future TTLL12-targeted intervention.