GLS-driven glutamine catabolism contributes to prostate cancer radiosensitivity by regulating the redox state, stemness and ATG5-mediated autophagy
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GLS-driven glutamine catabolism contributes to prostate cancer radiosensitivity by regulating the redox state, stemness and ATG5-mediated autophagy. / Mukha, Anna; Kahya, Uğur; Linge, Annett; Chen, Oleg; Löck, Steffen; Lukiyanchuk, Vasyl; Richter, Susan; Alves, Tiago C; Peitzsch, Mirko; Telychko, Vladyslav; Skvortsov, Sergej; Negro, Giulia; Aschenbrenner, Bertram; Skvortsova, Ira-Ida; Mirtschink, Peter; Lohaus, Fabian; Hölscher, Tobias; Neubauer, Hans; Rivandi, Mahdi; Labitzky, Vera; Lange, Tobias; Franken, André; Behrens, Bianca; Stoecklein, Nikolas H; Toma, Marieta; Sommer, Ulrich; Zschaeck, Sebastian; Rehm, Maximilian; Eisenhofer, Graeme; Schwager, Christian; Abdollahi, Amir; Groeben, Christer; Kunz-Schughart, Leoni A; Baretton, Gustavo B; Baumann, Michael; Krause, Mechthild; Peitzsch, Claudia; Dubrovska, Anna.
in: THERANOSTICS, Jahrgang 11, Nr. 16, 2021, S. 7844-7868.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - GLS-driven glutamine catabolism contributes to prostate cancer radiosensitivity by regulating the redox state, stemness and ATG5-mediated autophagy
AU - Mukha, Anna
AU - Kahya, Uğur
AU - Linge, Annett
AU - Chen, Oleg
AU - Löck, Steffen
AU - Lukiyanchuk, Vasyl
AU - Richter, Susan
AU - Alves, Tiago C
AU - Peitzsch, Mirko
AU - Telychko, Vladyslav
AU - Skvortsov, Sergej
AU - Negro, Giulia
AU - Aschenbrenner, Bertram
AU - Skvortsova, Ira-Ida
AU - Mirtschink, Peter
AU - Lohaus, Fabian
AU - Hölscher, Tobias
AU - Neubauer, Hans
AU - Rivandi, Mahdi
AU - Labitzky, Vera
AU - Lange, Tobias
AU - Franken, André
AU - Behrens, Bianca
AU - Stoecklein, Nikolas H
AU - Toma, Marieta
AU - Sommer, Ulrich
AU - Zschaeck, Sebastian
AU - Rehm, Maximilian
AU - Eisenhofer, Graeme
AU - Schwager, Christian
AU - Abdollahi, Amir
AU - Groeben, Christer
AU - Kunz-Schughart, Leoni A
AU - Baretton, Gustavo B
AU - Baumann, Michael
AU - Krause, Mechthild
AU - Peitzsch, Claudia
AU - Dubrovska, Anna
N1 - © The author(s).
PY - 2021
Y1 - 2021
N2 - Radiotherapy is one of the curative treatment options for localized prostate cancer (PCa). The curative potential of radiotherapy is mediated by irradiation-induced oxidative stress and DNA damage in tumor cells. However, PCa radiocurability can be impeded by tumor resistance mechanisms and normal tissue toxicity. Metabolic reprogramming is one of the major hallmarks of tumor progression and therapy resistance. Specific metabolic features of PCa might serve as therapeutic targets for tumor radiosensitization and as biomarkers for identifying the patients most likely to respond to radiotherapy. The study aimed to characterize a potential role of glutaminase (GLS)-driven glutamine catabolism as a prognostic biomarker and a therapeutic target for PCa radiosensitization. Methods: We analyzed primary cell cultures and radioresistant (RR) derivatives of the conventional PCa cell lines by gene expression and metabolic assays to identify the molecular traits associated with radiation resistance. Relative radiosensitivity of the cell lines and primary cell cultures were analyzed by 2-D and 3-D clonogenic analyses. Targeting of glutamine (Gln) metabolism was achieved by Gln starvation, gene knockdown, and chemical inhibition. Activation of the DNA damage response (DDR) and autophagy was assessed by gene expression, western blotting, and fluorescence microscopy. Reactive oxygen species (ROS) and the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) were analyzed by fluorescence and luminescence probes, respectively. Cancer stem cell (CSC) properties were investigated by sphere-forming assay, CSC marker analysis, and in vivo limiting dilution assays. Single circulating tumor cells (CTCs) isolated from the blood of PCa patients were analyzed by array comparative genome hybridization. Expression levels of the GLS1 and MYC gene in tumor tissues and amino acid concentrations in blood plasma were correlated to a progression-free survival in PCa patients. Results: Here, we found that radioresistant PCa cells and prostate CSCs have a high glutamine demand. GLS-driven catabolism of glutamine serves not only for energy production but also for the maintenance of the redox state. Consequently, glutamine depletion or inhibition of critical regulators of glutamine utilization, such as GLS and the transcription factor MYC results in PCa radiosensitization. On the contrary, we found that a combination of glutamine metabolism inhibitors with irradiation does not cause toxic effects on nonmalignant prostate cells. Glutamine catabolism contributes to the maintenance of CSCs through regulation of the alpha-ketoglutarate (α-KG)-dependent chromatin-modifying dioxygenase. The lack of glutamine results in the inhibition of CSCs with a high aldehyde dehydrogenase (ALDH) activity, decreases the frequency of the CSC populations in vivo and reduces tumor formation in xenograft mouse models. Moreover, this study shows that activation of the ATG5-mediated autophagy in response to a lack of glutamine is a tumor survival strategy to withstand radiation-mediated cell damage. In combination with autophagy inhibition, the blockade of glutamine metabolism might be a promising strategy for PCa radiosensitization. High blood levels of glutamine in PCa patients significantly correlate with a shorter prostate-specific antigen (PSA) doubling time. Furthermore, high expression of critical regulators of glutamine metabolism, GLS1 and MYC, is significantly associated with a decreased progression-free survival in PCa patients treated with radiotherapy. Conclusions: Our findings demonstrate that GLS-driven glutaminolysis is a prognostic biomarker and therapeutic target for PCa radiosensitization.
AB - Radiotherapy is one of the curative treatment options for localized prostate cancer (PCa). The curative potential of radiotherapy is mediated by irradiation-induced oxidative stress and DNA damage in tumor cells. However, PCa radiocurability can be impeded by tumor resistance mechanisms and normal tissue toxicity. Metabolic reprogramming is one of the major hallmarks of tumor progression and therapy resistance. Specific metabolic features of PCa might serve as therapeutic targets for tumor radiosensitization and as biomarkers for identifying the patients most likely to respond to radiotherapy. The study aimed to characterize a potential role of glutaminase (GLS)-driven glutamine catabolism as a prognostic biomarker and a therapeutic target for PCa radiosensitization. Methods: We analyzed primary cell cultures and radioresistant (RR) derivatives of the conventional PCa cell lines by gene expression and metabolic assays to identify the molecular traits associated with radiation resistance. Relative radiosensitivity of the cell lines and primary cell cultures were analyzed by 2-D and 3-D clonogenic analyses. Targeting of glutamine (Gln) metabolism was achieved by Gln starvation, gene knockdown, and chemical inhibition. Activation of the DNA damage response (DDR) and autophagy was assessed by gene expression, western blotting, and fluorescence microscopy. Reactive oxygen species (ROS) and the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) were analyzed by fluorescence and luminescence probes, respectively. Cancer stem cell (CSC) properties were investigated by sphere-forming assay, CSC marker analysis, and in vivo limiting dilution assays. Single circulating tumor cells (CTCs) isolated from the blood of PCa patients were analyzed by array comparative genome hybridization. Expression levels of the GLS1 and MYC gene in tumor tissues and amino acid concentrations in blood plasma were correlated to a progression-free survival in PCa patients. Results: Here, we found that radioresistant PCa cells and prostate CSCs have a high glutamine demand. GLS-driven catabolism of glutamine serves not only for energy production but also for the maintenance of the redox state. Consequently, glutamine depletion or inhibition of critical regulators of glutamine utilization, such as GLS and the transcription factor MYC results in PCa radiosensitization. On the contrary, we found that a combination of glutamine metabolism inhibitors with irradiation does not cause toxic effects on nonmalignant prostate cells. Glutamine catabolism contributes to the maintenance of CSCs through regulation of the alpha-ketoglutarate (α-KG)-dependent chromatin-modifying dioxygenase. The lack of glutamine results in the inhibition of CSCs with a high aldehyde dehydrogenase (ALDH) activity, decreases the frequency of the CSC populations in vivo and reduces tumor formation in xenograft mouse models. Moreover, this study shows that activation of the ATG5-mediated autophagy in response to a lack of glutamine is a tumor survival strategy to withstand radiation-mediated cell damage. In combination with autophagy inhibition, the blockade of glutamine metabolism might be a promising strategy for PCa radiosensitization. High blood levels of glutamine in PCa patients significantly correlate with a shorter prostate-specific antigen (PSA) doubling time. Furthermore, high expression of critical regulators of glutamine metabolism, GLS1 and MYC, is significantly associated with a decreased progression-free survival in PCa patients treated with radiotherapy. Conclusions: Our findings demonstrate that GLS-driven glutaminolysis is a prognostic biomarker and therapeutic target for PCa radiosensitization.
KW - Animals
KW - Autophagy
KW - Autophagy-Related Protein 5/metabolism
KW - Biomarkers, Pharmacological
KW - Cell Line, Tumor
KW - Glutaminase/antagonists & inhibitors
KW - Glutamine/metabolism
KW - Humans
KW - Male
KW - Mice, Nude
KW - Neoplastic Stem Cells/metabolism
KW - Oxidation-Reduction
KW - Prostatic Neoplasms/metabolism
KW - Proto-Oncogene Proteins c-myc/metabolism
KW - Radiation Tolerance/genetics
KW - Reactive Oxygen Species/metabolism
KW - Xenograft Model Antitumor Assays
U2 - 10.7150/thno.58655
DO - 10.7150/thno.58655
M3 - SCORING: Journal article
C2 - 34335968
VL - 11
SP - 7844
EP - 7868
JO - THERANOSTICS
JF - THERANOSTICS
SN - 1838-7640
IS - 16
ER -