Metabolic requirement for GOT2 in pancreatic cancer depends on environmental context
Mitochondrial glutamate-oxaloacetate transaminase 2 (GOT2) is part of the malate-aspartate shuttle, a mechanism by which cells transfer reducing equivalents from the cytosol to the mitochondria. GOT2 is a key component of mutant KRAS (KRAS*)-mediated rewiring of glutamine metabolism in pancreatic du...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-07-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/73245 |
| _version_ | 1828374320714350592 |
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| author | Samuel A Kerk Lin Lin Amy L Myers Damien J Sutton Anthony Andren Peter Sajjakulnukit Li Zhang Yaqing Zhang Jennifer A Jiménez Barbara S Nelson Brandon Chen Anthony Robinson Galloway Thurston Samantha B Kemp Nina G Steele Megan T Hoffman Hui-Ju Wen Daniel Long Sarah E Ackenhusen Johanna Ramos Xiaohua Gao Zeribe C Nwosu Stefanie Galban Christopher J Halbrook David B Lombard David R Piwnica-Worms Haoqiang Ying Marina Pasca di Magliano Howard C Crawford Yatrik M Shah Costas A Lyssiotis |
| author_facet | Samuel A Kerk Lin Lin Amy L Myers Damien J Sutton Anthony Andren Peter Sajjakulnukit Li Zhang Yaqing Zhang Jennifer A Jiménez Barbara S Nelson Brandon Chen Anthony Robinson Galloway Thurston Samantha B Kemp Nina G Steele Megan T Hoffman Hui-Ju Wen Daniel Long Sarah E Ackenhusen Johanna Ramos Xiaohua Gao Zeribe C Nwosu Stefanie Galban Christopher J Halbrook David B Lombard David R Piwnica-Worms Haoqiang Ying Marina Pasca di Magliano Howard C Crawford Yatrik M Shah Costas A Lyssiotis |
| author_sort | Samuel A Kerk |
| collection | DOAJ |
| description | Mitochondrial glutamate-oxaloacetate transaminase 2 (GOT2) is part of the malate-aspartate shuttle, a mechanism by which cells transfer reducing equivalents from the cytosol to the mitochondria. GOT2 is a key component of mutant KRAS (KRAS*)-mediated rewiring of glutamine metabolism in pancreatic ductal adenocarcinoma (PDA). Here, we demonstrate that the loss of GOT2 disturbs redox homeostasis and halts proliferation of PDA cells in vitro. GOT2 knockdown (KD) in PDA cell lines in vitro induced NADH accumulation, decreased Asp and α-ketoglutarate (αKG) production, stalled glycolysis, disrupted the TCA cycle, and impaired proliferation. Oxidizing NADH through chemical or genetic means resolved the redox imbalance induced by GOT2 KD, permitting sustained proliferation. Despite a strong in vitro inhibitory phenotype, loss of GOT2 had no effect on tumor growth in xenograft PDA or autochthonous mouse models. We show that cancer-associated fibroblasts (CAFs), a major component of the pancreatic tumor microenvironment (TME), release the redox active metabolite pyruvate, and culturing GOT2 KD cells in CAF conditioned media (CM) rescued proliferation in vitro. Furthermore, blocking pyruvate import or pyruvate-to-lactate reduction prevented rescue of GOT2 KD in vitro by exogenous pyruvate or CAF CM. However, these interventions failed to sensitize xenografts to GOT2 KD in vivo, demonstrating the remarkable plasticity and differential metabolism deployed by PDA cells in vitro and in vivo. This emphasizes how the environmental context of distinct pre-clinical models impacts both cell-intrinsic metabolic rewiring and metabolic crosstalk with the TME. |
| first_indexed | 2024-04-14T07:31:53Z |
| format | Article |
| id | doaj.art-3f6b7faa3ebf44d494617d12f6cd6c1c |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:31:53Z |
| publishDate | 2022-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-3f6b7faa3ebf44d494617d12f6cd6c1c2022-12-22T02:05:50ZengeLife Sciences Publications LtdeLife2050-084X2022-07-011110.7554/eLife.73245Metabolic requirement for GOT2 in pancreatic cancer depends on environmental contextSamuel A Kerk0https://orcid.org/0000-0001-9786-2245Lin Lin1Amy L Myers2Damien J Sutton3Anthony Andren4Peter Sajjakulnukit5Li Zhang6Yaqing Zhang7Jennifer A Jiménez8Barbara S Nelson9Brandon Chen10Anthony Robinson11Galloway Thurston12Samantha B Kemp13Nina G Steele14Megan T Hoffman15Hui-Ju Wen16Daniel Long17Sarah E Ackenhusen18Johanna Ramos19Xiaohua Gao20Zeribe C Nwosu21Stefanie Galban22Christopher J Halbrook23David B Lombard24David R Piwnica-Worms25Haoqiang Ying26Marina Pasca di Magliano27https://orcid.org/0000-0001-9632-9035Howard C Crawford28Yatrik M Shah29Costas A Lyssiotis30https://orcid.org/0000-0001-9309-6141Doctoral Program in Cancer Biology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDoctoral Program in Cancer Biology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Surgery, University of Michigan-Ann Arbor, Ann Arbor, United StatesDoctoral Program in Cancer Biology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDoctoral Program in Cancer Biology, University of Michigan-Ann Arbor, Ann Arbor, United States; Department of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesMolecular and Cellular Pathology Graduate Program, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesProgram in Chemical Biology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Radiology, University of Michigan, Ann Arbor, United States; Rogel Cancer Center, University of Michigan, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United StatesDepartment of Pathology and Institute of Gerontology, University of Michigan, Ann Arbor, United StatesDepartment of Cancer Systems Imaging, University of Texas MD Anderson Cancer Center, Houston, United StatesDepartment of Molecular and Cellular Oncology, University of Texas MD Anderson Cancer Center, Houston, United StatesDepartment of Surgery, University of Michigan-Ann Arbor, Ann Arbor, United States; Rogel Cancer Center, University of Michigan, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United States; Rogel Cancer Center, University of Michigan, Ann Arbor, United StatesDepartment of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United States; Rogel Cancer Center, University of Michigan, Ann Arbor, United States; Department of Internal Medicine, Division of Gastroenterology and Hepatology, University of Michigan, Ann Arbor, United StatesDoctoral Program in Cancer Biology, University of Michigan-Ann Arbor, Ann Arbor, United States; Department of Molecular and Integrative Physiology, University of Michigan-Ann Arbor, Ann Arbor, United States; Rogel Cancer Center, University of Michigan, Ann Arbor, United States; Department of Internal Medicine, Division of Gastroenterology and Hepatology, University of Michigan, Ann Arbor, United StatesMitochondrial glutamate-oxaloacetate transaminase 2 (GOT2) is part of the malate-aspartate shuttle, a mechanism by which cells transfer reducing equivalents from the cytosol to the mitochondria. GOT2 is a key component of mutant KRAS (KRAS*)-mediated rewiring of glutamine metabolism in pancreatic ductal adenocarcinoma (PDA). Here, we demonstrate that the loss of GOT2 disturbs redox homeostasis and halts proliferation of PDA cells in vitro. GOT2 knockdown (KD) in PDA cell lines in vitro induced NADH accumulation, decreased Asp and α-ketoglutarate (αKG) production, stalled glycolysis, disrupted the TCA cycle, and impaired proliferation. Oxidizing NADH through chemical or genetic means resolved the redox imbalance induced by GOT2 KD, permitting sustained proliferation. Despite a strong in vitro inhibitory phenotype, loss of GOT2 had no effect on tumor growth in xenograft PDA or autochthonous mouse models. We show that cancer-associated fibroblasts (CAFs), a major component of the pancreatic tumor microenvironment (TME), release the redox active metabolite pyruvate, and culturing GOT2 KD cells in CAF conditioned media (CM) rescued proliferation in vitro. Furthermore, blocking pyruvate import or pyruvate-to-lactate reduction prevented rescue of GOT2 KD in vitro by exogenous pyruvate or CAF CM. However, these interventions failed to sensitize xenografts to GOT2 KD in vivo, demonstrating the remarkable plasticity and differential metabolism deployed by PDA cells in vitro and in vivo. This emphasizes how the environmental context of distinct pre-clinical models impacts both cell-intrinsic metabolic rewiring and metabolic crosstalk with the TME.https://elifesciences.org/articles/73245tumor metabolismpancreatic cancertumor microenvironment |
| spellingShingle | Samuel A Kerk Lin Lin Amy L Myers Damien J Sutton Anthony Andren Peter Sajjakulnukit Li Zhang Yaqing Zhang Jennifer A Jiménez Barbara S Nelson Brandon Chen Anthony Robinson Galloway Thurston Samantha B Kemp Nina G Steele Megan T Hoffman Hui-Ju Wen Daniel Long Sarah E Ackenhusen Johanna Ramos Xiaohua Gao Zeribe C Nwosu Stefanie Galban Christopher J Halbrook David B Lombard David R Piwnica-Worms Haoqiang Ying Marina Pasca di Magliano Howard C Crawford Yatrik M Shah Costas A Lyssiotis Metabolic requirement for GOT2 in pancreatic cancer depends on environmental context eLife tumor metabolism pancreatic cancer tumor microenvironment |
| title | Metabolic requirement for GOT2 in pancreatic cancer depends on environmental context |
| title_full | Metabolic requirement for GOT2 in pancreatic cancer depends on environmental context |
| title_fullStr | Metabolic requirement for GOT2 in pancreatic cancer depends on environmental context |
| title_full_unstemmed | Metabolic requirement for GOT2 in pancreatic cancer depends on environmental context |
| title_short | Metabolic requirement for GOT2 in pancreatic cancer depends on environmental context |
| title_sort | metabolic requirement for got2 in pancreatic cancer depends on environmental context |
| topic | tumor metabolism pancreatic cancer tumor microenvironment |
| url | https://elifesciences.org/articles/73245 |
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