Disruption of redox balance in glutaminolytic triple negative breast cancer by inhibition of glutamate export and glutaminase

Abstract

In triple-negative breast cancer (TNBC) that relies on catabolism of amino acid glutamine, glutaminase (GLS) converts glutamine to glutamate, which facilitates glutathione synthesis by mediating the enrichment of intracellular cystine via xCT antiporter activity. To overcome chemo resistant TNBC, we have tested a strategy of disrupting cellular redox balance by inhibition of GLS and xCT by CB839 and Erastin, respectively. Key findings of our study include: 1. Dual metabolic inhibition (CB839+Erastin) led to significant increases of cellular superoxide level in both parent and chemo resistant TNBC cells, but superoxide level was distinctly lower in resistant cells. 2. Dual metabolic inhibition combined with doxorubicin or cisplatin induced significant apoptosis in TNBC cells and is associated with high degrees of GSH depletion.In vivo, dual metabolic inhibition plus cisplatin led to significant growth delay of chemo resistant human TNBC xenografts. 3. Ferroptosis is induced by doxorubicin (DOX) but not by cisplatin or paclitaxel. Addition of dual metabolic inhibition to DOX chemotherapy significantly enhanced ferroptotic cell death. 4. Significant changes in cellular metabolites concentration preceded transcriptome changes revealed by single cell RNA sequencing, underscoring the potential of capturing early changes in metabolites as pharmacodynamic markers of metabolic inhibitors. Here we demonstrated that 4-(3-[18F]fluoropropyl)-L-glutamic acid ([18F]FSPG) PET detected xCT blockade by Erastin or its analog in mice bearing human TNBC xenografts. In summary, our study provides compelling evidence for the therapeutic benefit and feasibility of non-invasive monitoring of dual metabolic blockade as a translational strategy to sensitize chemo resistant TNBC to cytotoxic chemotherapy.

GLS and GLS2 Glutaminase Isoenzymes in the Antioxidant System of Cancer Cells

“…Ferroptosis modulation is essential in the chemoresistance event in TNBC [ 99 ], whereby tumor cells are characterized as commonly being Gln-dependent via glutaminolysis, which favors drug resistance [ 100 ]. To defeat these cancer cells, a dual combination strategy has been suggested by targeting both GLS and ferroptosis [ 99 ].…”

“…Ferroptosis modulation is essential in the chemoresistance event in TNBC [ 99 ], whereby tumor cells are characterized as commonly being Gln-dependent via glutaminolysis, which favors drug resistance [ 100 ]. To defeat these cancer cells, a dual combination strategy has been suggested by targeting both GLS and ferroptosis [ 99 ]. Gallbladder cancer cells, which also need GLS for growth and proliferation, triggered ferroptosis following GLS inhibition and a drop in antioxidant capacity (GSH- and NADPH-dependent), which evoked oxidative stress [ 101 ].…”