ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors
Abstract Background ATM is a multifunctional serine/threonine kinase that in addition to its well-established role in DNA repair mechanisms is involved in a number of signaling pathways including regulation of oxidative stress response and metabolic diversion of glucose through the pentose phosphate...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-11-01
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| Series: | Cancer & Metabolism |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40170-023-00320-4 |
| _version_ | 1827764248178065408 |
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| author | Cristina Terlizzi Viviana De Rosa Francesca Iommelli Antonio Pezone Giovanna G. Altobelli Maurizio Maddalena Jelena Dimitrov Caterina De Rosa Carminia Maria Della Corte Vittorio Enrico Avvedimento Silvana Del Vecchio |
| author_facet | Cristina Terlizzi Viviana De Rosa Francesca Iommelli Antonio Pezone Giovanna G. Altobelli Maurizio Maddalena Jelena Dimitrov Caterina De Rosa Carminia Maria Della Corte Vittorio Enrico Avvedimento Silvana Del Vecchio |
| author_sort | Cristina Terlizzi |
| collection | DOAJ |
| description | Abstract Background ATM is a multifunctional serine/threonine kinase that in addition to its well-established role in DNA repair mechanisms is involved in a number of signaling pathways including regulation of oxidative stress response and metabolic diversion of glucose through the pentose phosphate pathway. Oncogene-driven tumorigenesis often implies the metabolic switch from oxidative phosphorylation to glycolysis which provides metabolic intermediates to sustain cell proliferation. The aim of our study is to elucidate the role of ATM in the regulation of glucose metabolism in oncogene-driven cancer cells and to test whether ATM may be a suitable target for anticancer therapy. Methods Two oncogene-driven NSCLC cell lines, namely H1975 and H1993 cells, were treated with ATM inhibitor, KU55933, alone or in combination with oncogene driver inhibitors, WZ4002 or crizotinib. Key glycolytic enzymes, mitochondrial complex subunits (OXPHOS), cyclin D1, and apoptotic markers were analyzed by Western blotting. Drug-induced toxicity was assessed by MTS assay using stand-alone or combined treatment with KU55933 and driver inhibitors. Glucose consumption, pyruvate, citrate, and succinate levels were also analyzed in response to KU55933 treatment. Both cell lines were transfected with ATM-targeted siRNA or non-targeting siRNA and then exposed to treatment with driver inhibitors. Results ATM inhibition deregulates and inhibits glucose metabolism by reducing HKII, p-PKM2Tyr105, p-PKM2Ser37, E1α subunit of pyruvate dehydrogenase complex, and all subunits of mitochondrial complexes except ATP synthase. Accordingly, glucose uptake and pyruvate concentrations were reduced in response to ATM inhibition, whereas citrate and succinate levels were increased in both cell lines indicating the supply of alternative metabolic substrates. Silencing of ATM resulted in similar changes in glycolytic cascade and OXPHOS levels. Furthermore, the driver inhibitors amplified the effects of ATM downregulation on glucose metabolism, and the combined treatment with ATM inhibitors enhanced the cytotoxic effect of driver inhibitors alone by increasing the apoptotic response. Conclusions Inhibition of ATM reduced both glycolytic enzymes and OXPHOS levels in oncogene-driven cancer cells and enhanced apoptosis induced by driver inhibitors thus highlighting the possibility to use ATM and the driver inhibitors in combined regimens of anticancer therapy in vivo. |
| first_indexed | 2024-03-11T11:01:40Z |
| format | Article |
| id | doaj.art-e83d3991835d4c898a45587c3d859097 |
| institution | Directory Open Access Journal |
| issn | 2049-3002 |
| language | English |
| last_indexed | 2024-03-11T11:01:40Z |
| publishDate | 2023-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer & Metabolism |
| spelling | doaj.art-e83d3991835d4c898a45587c3d8590972023-11-12T12:27:46ZengBMCCancer & Metabolism2049-30022023-11-0111111110.1186/s40170-023-00320-4ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitorsCristina Terlizzi0Viviana De Rosa1Francesca Iommelli2Antonio Pezone3Giovanna G. Altobelli4Maurizio Maddalena5Jelena Dimitrov6Caterina De Rosa7Carminia Maria Della Corte8Vittorio Enrico Avvedimento9Silvana Del Vecchio10Department of Advanced Biomedical Sciences, University “Federico II”Institute of Biostructures and Bioimaging, National Research CouncilInstitute of Biostructures and Bioimaging, National Research CouncilDepartment of Biology, University “Federico II”Department of Advanced Biomedical Sciences, University “Federico II”Department of Advanced Biomedical Sciences, University “Federico II”Department of Advanced Biomedical Sciences, University “Federico II”Department of Precision Medicine, University of Campania Luigi VanvitelliDepartment of Precision Medicine, University of Campania Luigi VanvitelliDepartment of Molecular Medicine and Medical Biotechnology, University “Federico II”Department of Advanced Biomedical Sciences, University “Federico II”Abstract Background ATM is a multifunctional serine/threonine kinase that in addition to its well-established role in DNA repair mechanisms is involved in a number of signaling pathways including regulation of oxidative stress response and metabolic diversion of glucose through the pentose phosphate pathway. Oncogene-driven tumorigenesis often implies the metabolic switch from oxidative phosphorylation to glycolysis which provides metabolic intermediates to sustain cell proliferation. The aim of our study is to elucidate the role of ATM in the regulation of glucose metabolism in oncogene-driven cancer cells and to test whether ATM may be a suitable target for anticancer therapy. Methods Two oncogene-driven NSCLC cell lines, namely H1975 and H1993 cells, were treated with ATM inhibitor, KU55933, alone or in combination with oncogene driver inhibitors, WZ4002 or crizotinib. Key glycolytic enzymes, mitochondrial complex subunits (OXPHOS), cyclin D1, and apoptotic markers were analyzed by Western blotting. Drug-induced toxicity was assessed by MTS assay using stand-alone or combined treatment with KU55933 and driver inhibitors. Glucose consumption, pyruvate, citrate, and succinate levels were also analyzed in response to KU55933 treatment. Both cell lines were transfected with ATM-targeted siRNA or non-targeting siRNA and then exposed to treatment with driver inhibitors. Results ATM inhibition deregulates and inhibits glucose metabolism by reducing HKII, p-PKM2Tyr105, p-PKM2Ser37, E1α subunit of pyruvate dehydrogenase complex, and all subunits of mitochondrial complexes except ATP synthase. Accordingly, glucose uptake and pyruvate concentrations were reduced in response to ATM inhibition, whereas citrate and succinate levels were increased in both cell lines indicating the supply of alternative metabolic substrates. Silencing of ATM resulted in similar changes in glycolytic cascade and OXPHOS levels. Furthermore, the driver inhibitors amplified the effects of ATM downregulation on glucose metabolism, and the combined treatment with ATM inhibitors enhanced the cytotoxic effect of driver inhibitors alone by increasing the apoptotic response. Conclusions Inhibition of ATM reduced both glycolytic enzymes and OXPHOS levels in oncogene-driven cancer cells and enhanced apoptosis induced by driver inhibitors thus highlighting the possibility to use ATM and the driver inhibitors in combined regimens of anticancer therapy in vivo.https://doi.org/10.1186/s40170-023-00320-4ATMGlycolysisOXPHOSOncogene-driver inhibitorsAnticancer therapy |
| spellingShingle | Cristina Terlizzi Viviana De Rosa Francesca Iommelli Antonio Pezone Giovanna G. Altobelli Maurizio Maddalena Jelena Dimitrov Caterina De Rosa Carminia Maria Della Corte Vittorio Enrico Avvedimento Silvana Del Vecchio ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors Cancer & Metabolism ATM Glycolysis OXPHOS Oncogene-driver inhibitors Anticancer therapy |
| title | ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors |
| title_full | ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors |
| title_fullStr | ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors |
| title_full_unstemmed | ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors |
| title_short | ATM inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors |
| title_sort | atm inhibition blocks glucose metabolism and amplifies the sensitivity of resistant lung cancer cell lines to oncogene driver inhibitors |
| topic | ATM Glycolysis OXPHOS Oncogene-driver inhibitors Anticancer therapy |
| url | https://doi.org/10.1186/s40170-023-00320-4 |
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