Transcriptional upregulation of NDUFA10 by ATF3 sustains mitochondrial bioenergetics and malignant phenotypes in NSCLC

Abstract Non-small cell lung cancer (NSCLC) remains a leading cause of global cancer mortality, necessitating the identification of novel therapeutic vulnerabilities. This study identifies NDUFA10, a key mitochondrial Complex I subunit, as a critical driver of NSCLC pathogenesis. Multi-cohort analysis of TCGA and clinical datasets revealed significant NDUFA10 upregulation in neoplastic tissues, which correlated with advanced pathological stage, metastasis, smoking history, and poor overall survival. Single-cell RNA sequencing further localized NDUFA10 enrichment to proliferating malignant epithelial subpopulations, where its expression positively associated with oxidative phosphorylation (OXPHOS) and ATP synthesis across primary and metastatic sites. Functionally, NDUFA10 ablation (shRNA or CRISPR/Cas9) in primary NSCLC and A549 cells disrupted mitochondrial homeostasis—evidenced by impaired respiration, attenuated Complex I activity, and augmented ROS production—while inhibiting cell viability, migration, and invasion. Mechanistically, NDUFA10 modulates the Akt-mTOR signaling axis; constitutively active Akt1 partially rescued the proliferative and survival deficits induced by NDUFA10 silencing. Furthermore, intersectional bioinformatics and ChIP-qPCR identified ATF3 as the direct transcriptional activator driving NDUFA10 overexpression. In vivo, NDUFA10-silenced xenografts exhibited significantly attenuated growth, characterized by compromised mitochondrial function and inhibited Akt-mTOR activation. In summary, our findings characterize NDUFA10 as a critical metabolic checkpoint in NSCLC, where its ATF3-driven its overexpression coordinates mitochondrial bioenergetics and Akt-mTOR signaling to sustain malignant progression.