Abstract 436: Extracellular glutathione catabolism as an alternative cyst(e)ine source in cancer

Abstract Glutathione (GSH) is the most abundant antioxidant in the human body and plays essential roles in physiology and disease. Unchecked oxidative stress is involved in tumor initiation and progression. Paradoxically, clinical and laboratory data show that ROS-scavenging molecules such as glutathione not only fail to impede tumor growth but, in many cases, accelerate it. Still, preliminary data from our lab show that genetically ablating tumors’ ability to synthesize GSH does not affect their growth. This finding suggests that the favorable influence of glutathione on tumor growth may arise from external sources. Yet, the mechanism cancer cells employ to take advantage of extracellular glutathione in this context is unknown. Here, we show that γ-glutamyl-transpeptidase 1 (GGT1), an enzyme that localizes on the outer side of the cell’s membrane, can break down extracellular GSH, yielding glutamate and the dipeptide cysteinylglycine. Cysteinylglycine can be further metabolized, generating cysteine and glycine to the cells. We found that the addition of exogenous GSH, as well as cysteinylglycine, can mitigate cell death, proliferation arrest, and oxidative stress caused by cystine depletion. The overexpression of human GGT1 in cancer cells significantly reduces the amount of glutathione needed to rescue cell fitness. Accordingly, pharmacologic inhibition of GGT1 activity prevented GSH's ability to rescue cell proliferation. Moreover, an unbiased pharmacological screening revealed that GGT1-mediated cysteine acquisition renders cancer cells resistant to inhibitors of cystine uptake and utilization, supporting the role of GGT1 as a source of cyst(e)ine. Our findings reveal that cancer cells can access cysteine, a vital yet scarce amino acid, through GGT1's ability to break down glutathione, which is present in remarkably high concentrations within the tumor microenvironment. Although the downstream proteins involved in the transport and utilization of GSH breakdown products are yet to be identified, we anticipate this pathway has the potential to uncover novel therapeutic avenues for cancer treatment by targeting alternative pathways of nutrient acquisition that are potentially implicated in cancer progression and therapy resistance. Citation Format: Fabio Hecht, Marco Zocchi, Emily Tuttle, Gloria Asantewaa, TashJaé Scales, Fatemeh Alimohammadi, Nathan Ward, Gina DeNicola, Isaac Harris. Extracellular glutathione catabolism as an alternative cyst(e)ine source in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 436.