Deubiquitinases Maintain Protein Homeostasis and Survival of Cancer Cells upon Glutathione Depletion

Harris, Isaac S.; Endress, Jennifer E.; Coloff, Jonathan L.; Selfors, Laura M.; McBrayer, Samuel K.; Rosenbluth, Jennifer M.; Takahashi, Nobuaki; Dhakal, Sabin; Koduri, Vidyasagar; Oser, Matthew G.; Schauer, Nathan J.; Doherty, Laura M.; Hong, Andrew L.; Kang, Yun Pyo; Younger, Scott T.; Doench, John G.; Hahn, William C.; Buhrlage, Sara J.; DeNicola, Gina M.; Kaelin, William G.; Brugge, Joan S.

doi:10.1016/j.cmet.2019.01.020

Cited by 138 publications

(129 citation statements)

References 110 publications

(111 reference statements)

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“…Figure 8D) did not induce lipid ROS ( Figure 3E) and had no impact on cell viability ( Figure 3F). This result highlights contradictory findings regarding the sufficiency of glutathione depletion for the induction of ferroptosis in different cell lines (13,30). We inferred that in PDA cells, the loss of one or more additional cysteine-derived metabolites may be critical to the induction of ferroptosis.…”

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confidence: 80%

“Induction of pancreatic tumor-selective ferroptosis through modulation of cystine import”

Badgley

Kremer

Maurer

et al. 2019

Preprint

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Pancreatic ductal adenocarcinoma (PDA) is the third-leading cause of cancer mortality in the US and is highly resistant to classical, targeted, and immune therapies. We show that human PDA cells are dependent on the provision of exogenous cystine to avert a catastrophic accumulation of lipid reactive oxygen species (ROS) that, left unchecked, leads to ferroptotic cell death, both in vitro and in vivo. Using a dual-recombinase genetically engineered model, we found that acute deletion of Slc7a11 led to tumorselective ferroptosis, tumor stabilizations/regressions, and extended overall survival. The mechanism of ferroptosis induction in PDA cells required the concerted depletion of both glutathione and coenzyme A, highlighting a novel branch of ferroptosis-relevant metabolism. Finally, we found that cystine depletion in vivo using the pre-IND agent cyst(e)inase phenocopied Slc7a11 deletion, inducing tumor-selective ferroptosis and disease stabilizations/regressions in the well-validated KPC model of PDA.One Sentence Summary: Genetic and pharmacological targeting of cystine import induces pancreatic cancer-selective ferroptosis in vivo. Main Body:The cancer-selective induction of apoptosis through cytotoxic chemotherapy is a foundation of modern oncology. However, some cancers, such as PDA, have proven highly resistant to traditional therapeutic strategies (1). In addition to proliferative signals, activating mutations in KRAS (found in >90% of human pancreatic tumors) induce both an increase in the generation of ROS as well as compensatory antioxidant programs (2-4). We hypothesize that in this state of elevated ROS flux, ROS detoxification programs become a critical metabolic dependency.Cellular ROS are neutralized primarily by thiols derived from the semi-essential amino acid cysteine (5-10). Antioxidant molecules such as glutathione and thioredoxin serve to position the uniquelyreactive sulfhydryl group of cysteine to catalyze ROS detoxification reactions. As cysteine is ultimately derived from extracellular sources, we examined the effects of culturing PDA cell lines in the absence of Author Contributions

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mentioning

confidence: 80%

“Induction of pancreatic tumor-selective ferroptosis through modulation of cystine import”

Badgley

Kremer

Maurer

et al. 2019

Preprint

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“…Recent studies have suggested that a majority of cancer cell lines survive upon 72 hours of BSO treatment despite potent inhibition of GSH levels at this time point. Moreover, these studies have demonstrated that chronic BSO treatment, up to 9 days, is required in induce cell death or stasis 27 . In our models, 6 hours of BSO treatment was sufficient to diminish GSH levels (Figure S3E) , in line with previous kinetic data 27 .…”

Section: Resultsmentioning

confidence: 99%

“…Moreover, these studies have demonstrated that chronic BSO treatment, up to 9 days, is required in induce cell death or stasis 27 . In our models, 6 hours of BSO treatment was sufficient to diminish GSH levels (Figure S3E) , in line with previous kinetic data 27 . By contrast, co-treatment of GOT1 knockdown with BSO was cytotoxic at 72 hours, while BSO alone was cytostatic (Figures 3D and S3F) .…”

Section: Resultsmentioning

confidence: 99%

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GOT1 Inhibition Primes Pancreatic Cancer for Ferroptosis through the Autophagic Release of Labile Iron

Kremer

Nelson

Lin

et al. 2020

Preprint

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“…These redox Western samples were mixed with a 4× nonreducing buffer before separation by SDS-PAGE. For analyses of TXN1 oxidation, a previously established, urea-PAGE protocol was followed (Du et al, 2013; Harris et al, 2019). Two control lysates were fully reduced with dithiothreitol (VWR) and then treated with iodoacetic acid (Sigma-Aldrich) or iodoacetamide (Sigma-Aldrich) to represent fully oxidized or reduced TXN protein.…”

Section: Methodsmentioning

confidence: 99%

Nicotinamide nucleotide transhydrogenase regulates mitochondrial metabolism in NSCLC through maintenance of Fe-S protein function

Ward

Kang

Falzone

et al. 2020

Journal of Experimental Medicine

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Human lung tumors exhibit robust and complex mitochondrial metabolism, likely precipitated by the highly oxygenated nature of pulmonary tissue. As ROS generation is a byproduct of this metabolism, reducing power in the form of nicotinamide adenine dinucleotide phosphate (NADPH) is required to mitigate oxidative stress in response to this heightened mitochondrial activity. Nicotinamide nucleotide transhydrogenase (NNT) is known to sustain mitochondrial antioxidant capacity through the generation of NADPH; however, its function in non-small cell lung cancer (NSCLC) has not been established. We found that NNT expression significantly enhances tumor formation and aggressiveness in mouse models of lung tumor initiation and progression. We further show that NNT loss elicits mitochondrial dysfunction independent of substantial increases in oxidative stress, but rather marked by the diminished activities of proteins dependent on resident iron-sulfur clusters. These defects were associated with both NADPH availability and ROS accumulation, suggesting that NNT serves a specific role in mitigating the oxidation of these critical protein cofactors.

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Deubiquitinases Maintain Protein Homeostasis and Survival of Cancer Cells upon Glutathione Depletion

Cited by 138 publications

References 110 publications

“Induction of pancreatic tumor-selective ferroptosis through modulation of cystine import”

“Induction of pancreatic tumor-selective ferroptosis through modulation of cystine import”

GOT1 Inhibition Primes Pancreatic Cancer for Ferroptosis through the Autophagic Release of Labile Iron

Nicotinamide nucleotide transhydrogenase regulates mitochondrial metabolism in NSCLC through maintenance of Fe-S protein function

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