YWHAG mediated epithelial-mesenchymal transition facilitates tumor progression and metastatic spread in non-small cell lung cancer.

BACKGROUND

YWHAG, a 14-3-3 protein family member, has been implicated in tumorigenesis, yet its role in lung cancer progression and underlying mechanisms remain poorly understood. This study aimed to elucidate the functional and mechanistic contributions of YWHAG to non-small cell lung cancer (NSCLC).

METHODS

Multi-omics analyses were performed using TCGA pan-cancer datasets, NSCLC single-cell RNA sequencing (scRNA-seq) data (GSE117570 & GSE148071), and public NSCLC mouse models. Functional validation included in vitro siRNA knockdown and overexpression of YWHAG in NCI-H292 cells, co-immunoprecipitation (Co-IP), immunofluorescence, and pathway activity assays. A lung-specific YWHAG knockout mouse model was generated using AAV5-SPC-Cre, and tumor growth/metastasis were assessed in an orthotopic A549 cell-derived NSCLC model via bioluminescence imaging and histopathology.

RESULTS

YWHAG was significantly upregulated across 29 cancer types, including lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC), and correlated with poor patient survival. scRNA-seq revealed YWHAG-enriched proliferating epithelial cells (ECs) with elevated EMT activity (EMT_AUC score), marked by upregulation of COL12A1, FN1, and YWHAG. Mechanistically, YWHAG knockdown suppressed EMT (increased E-cadherin, decreased N-cadherin/Vimentin), proliferation, and migration, while overexpression exerted opposite effects. YWHAG interacted with LRRK2 to activate the PI3K/AKT pathway, and LRRK2 overexpression rescued PI3K/AKT suppression post-YWHAG knockdown. In mice, lung-specific YWHAG knockout attenuated tumor growth and metastasis, suppressed EMT markers, and reduced metastatic burden in lung, liver, and skeletal muscle.

CONCLUSION

YWHAG drives NSCLC progression by promoting EMT, proliferation, and metastasis via the LRRK2/PI3K/AKT axis. Its pan-cancer overexpression and prognostic significance highlights YWHAG as a promising therapeutic target in lung cancer.