GLS2 links glutamine metabolism and atherosclerosis by remodelling artery walls (MOVAS).

Metabolic dysregulation, including perturbed glutamine-glutamate homeostasis, is common in atherosclerotic cardiovascular disease. Here, we reveal that modulation of glutaminolysis or glutamate availability in culture media is critical for smooth muscle cell line (MOVAS) phenotypic switching. Cells were treated for 24 hours with glutaminase inhibitors (50mM DON, Sigma-SML0601) in presence or absence of 2mM glutamate suplementation (Gibco-35050061). High-throughput transcriptional profiling revealed that modulation of glutamine and glutamate homeostasis impacted genes involved in protein and extracellular matrix organization, cell motility and microtubule. Thus, we uncovered a novel role for glutamine metabolism in the phenotypic switch in SMCs, a hallmark of vascular remodelling. Immortalized smooth muscle cell line MOVAS (ATCC CRL-2797) were cultured at 37°C and 5% CO2 in standatd growth media consisting of Dulbecco’s Modified Eagle’s Medium (DMEM, Corning), 100 U/ml of penicillin/streptomycin (Corning), 10% Fetal Bovine Serum (Corning) and 0.2mg/mL geneticin (G418, VWR Life Science). Cells used at 60-70% confluency were treated for 24 hours with glutaminase inhibitors (50mM DON, Sigma-SML0601) and supplemented or not with 2mM glutamate (Gibco-35050061)