Lactylation of ENO1 drives lung adenocarcinoma progression and resistance to EGFR-TKIs by stabilizing YAP1

Targeted tyrosine kinase inhibitors (TKIs) have achieved remarkable clinical efficacy in lung adenocarcinoma (LUAD); however, acquired resistance inevitably limits long-term treatment benefit. This study aimed to investigate the role of ENO1 in LUAD progression and TKI resistance, and to elucidate the underlying molecular mechanisms. ENO1 expression was analyzed in LUAD tissues and cell lines using TCGA data, qPCR, western blot, and immunohistochemistry. Functional assays, including CCK-8, colony formation, subcutaneous xenograft models, and a CRISPR/Cas9-based autochthonous GEMM, were performed to assess the effects of ENO1 modulation on LUAD cell proliferation and tumor growth. Mechanistically, RNA sequencing, co-immunoprecipitation, ubiquitination assays, and mass spectrometry were employed to identify ENO1-interacting proteins and post-translational modifications. ENO1 was significantly upregulated at both mRNA and protein levels in LUAD, and high ENO1 expression was closely associated with poor patient prognosis. ENO1 deficiency markedly inhibited LUAD cell proliferation in vitro and tumor growth in vivo. Mechanistically, ENO1 directly interacted with YAP1 and prevented its ubiquitination and proteasome-mediated degradation, independent of canonical Hippo kinases. Notably, lactylation of ENO1 at lysine 420 (K420) enhanced its interaction with YAP1. Elevated ENO1 K420 lactylation was observed in osimertinib-resistant PC-9 and H1975 cells, contributing to YAP1 hyperactivation and drug resistance. Importantly, targeting ENO1 effectively restored the sensitivity of resistant LUAD cells to osimertinib. Our findings demonstrate that ENO1 K420 lactylation serves as a key post-translational modification that stabilizes YAP1, drives LUAD progression and EGFR-TKI resistance, and implicate the ENO1-YAP1 axis as a promising candidate therapeutic target for LUAD. Not applicable.