TY - JOUR
T1 - Metabolic Requirement for GOT2 in Pancreatic Cancer Depends on Environmental Context
AU - Kerk, Samuel A.
AU - Lin, Lin
AU - Myers, Amy L.
AU - Sutton, Damien J.
AU - Andren, Anthony
AU - Sajjakulnukit, Peter
AU - Zhang, Li
AU - Zhang, Yaqing
AU - Jiménez, Jennifer A.
AU - Nelson, Barbara S.
AU - Chen, Brandon
AU - Robinson, Anthony
AU - Thurston, Galloway
AU - Kemp, Samantha B.
AU - Steele, Nina G.
AU - Hoffman, Megan T.
AU - Wen, Hui Ju
AU - Long, Daniel
AU - Ackenhusen, Sarah E.
AU - Ramos, Johanna
AU - Gao, Xiaohua
AU - Nwosu, Zeribe C.
AU - Galbán, Stefanie
AU - Halbrook, Christopher J.
AU - Lombard, David B.
AU - Piwnica-Worms, David R.
AU - Ying, Haoqiang
AU - Magliano, Marina Pasca Di
AU - Crawford, Howard C.
AU - Shah, Yatrik M.
AU - Lyssiotis, Costas A.
N1 - Publisher Copyright:
© 2022, eLife Sciences Publications Ltd. All rights reserved.
PY - 2022/7
Y1 - 2022/7
N2 - Mitochondrial glutamate-oxaloacetate (GOT2) is part of the malate-aspartate shuttle (MAS), 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 tumor microenvironment (TME).
AB - Mitochondrial glutamate-oxaloacetate (GOT2) is part of the malate-aspartate shuttle (MAS), 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 tumor microenvironment (TME).
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U2 - 10.7554/elife.73245
DO - 10.7554/elife.73245
M3 - Article
C2 - 35815941
AN - SCOPUS:85133891233
SN - 2050-084X
VL - 11
JO - eLife
JF - eLife
M1 - e73245
ER -