Patient behavior and physiology can directly affect cancer metabolism. Smoking is the leading risk factor for non-small cell lung cancer (NSCLC). Here, we identified that smoking modulates lung cancer cell metabolism through altered protein post-translational modification. Proteomic analyses identified elevated K251 succinylation (K251-Su) of GAPDH, a key enzyme in glycolysis, in NSCLC samples, and GAPDH K251-Su was significantly higher in patients who smoke compared to non-smokers. Exposure of lung cancer cells to cigarette smoke extract led to increased uptake of glutamine and enhanced GAPDH K251-Su. Glutamine uptake by cancer cells in hypoxic and nutrient-deficient microenvironments provided succinyl-CoA donors for GAPDH succinylation at K251, which was catalyzed by acyltransferase p300. K251-Su increased GAPDH stability by suppressing TRIM4-mediated K254 ubiquitination. GAPDH K251-Su enhanced glycolysis and glutamine reductive carboxylation to meet the demands for cell growth and to support survival in hypoxic and nutrient-depleted conditions, promoting tumor growth and metastasis. These findings indicate that tobacco smoking mediates metabolic reprogramming of cancer cells through succinylation of GAPDH to drive NSCLC progression.

The positive rate of GAPDH K251-Su in the tumor and adjacent normal tissue of NSCLC

The 3D reconstruction video of lung micro-CT scanning in a KrasG12D-driven NSCLC GAPDH WT mouse. The red nodular lesions in the 3D reconstructed video indicate tumor lesions.

Supplementary Figure 6 (Related to Fig.3).

The effects of cigarette smoke extract and various amino acids on GAPDH K251-Su, GAPDH and glutamine uptake in A549 cells under different culture conditions.

The 3D reconstruction video of lung micro-CT scanning in a KrasG12D-driven NSCLC GAPDH K251R mouse. The red nodular lesions in the 3D reconstructed video indicate tumor lesions.