Severe metabolic alterations in liver cancer lead to ERK pathway activation and drug resistance
Zeribe Chike Nwosu,
Weronika Piorońska,
Nadia Battello,
Andreas David Zimmer,
Bedair Dewidar,
Mei Han,
Sharon Pereira,
Biljana Blagojevic,
Darko Castven,
Verodia Charlestin,
Pavlo Holenya,
Julia Lochead,
Carolina De La Torre,
Norbert Gretz,
Peter Sajjakulnukit,
Li Zhang,
Matthew H. Ward,
Jens U. Marquardt,
Marina Pasca di Magliano,
Costas A. Lyssiotis,
Jonathan Sleeman,
Stefan Wölfl,
Matthias Philip Ebert,
Christoph Meyer,
Ute Hofmann,
Steven Dooley
Affiliations
Zeribe Chike Nwosu
Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany; Current Address of Corresponding author: Department of Molecular and Integrative Physiology, Rogel Cancer Centre, University of Michigan, Ann Arbor, 48109 MI, USA.
Weronika Piorońska
Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany
Nadia Battello
Luxembourg Science Center, L-4620 Differdange, Luxembourg, Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4362 Esch-Belval, Luxembourg
Andreas David Zimmer
Signal Transduction Laboratory, Life Sciences Research Unit, University of Luxembourg, L-4367 Belvaux, Luxembourg
Bedair Dewidar
Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tanta University, 31527 Tanta, Egypt
Mei Han
Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany
Sharon Pereira
Department of Medicine I, Lichtenberg Research Group, Johannes Gutenberg University, Mainz, Germany
Biljana Blagojevic
Department of Biology, Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, 69120 Heidelberg, Germany
Darko Castven
Department of Medicine I, Lichtenberg Research Group, Johannes Gutenberg University, Mainz, Germany
Verodia Charlestin
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, 46556 IN, United States
Pavlo Holenya
Department of Biology, Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, 69120 Heidelberg, Germany
Julia Lochead
Department of Biology, Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, 69120 Heidelberg, Germany
Carolina De La Torre
Medical Research Center, Medical Faculty of Mannheim, University of Heidelberg, Mannheim, Germany
Norbert Gretz
Medical Research Center, Medical Faculty of Mannheim, University of Heidelberg, Mannheim, Germany
Peter Sajjakulnukit
Rogel Cancer Center, University of Michigan, Ann Arbor 48109 MI, United States
Li Zhang
Rogel Cancer Center, University of Michigan, Ann Arbor 48109 MI, United States
Matthew H. Ward
Rogel Cancer Center, University of Michigan, Ann Arbor 48109 MI, United States
Jens U. Marquardt
Department of Medicine I, Lichtenberg Research Group, Johannes Gutenberg University, Mainz, Germany
Marina Pasca di Magliano
Rogel Cancer Center, University of Michigan, Ann Arbor 48109 MI, United States
Costas A. Lyssiotis
Rogel Cancer Center, University of Michigan, Ann Arbor 48109 MI, United States
Jonathan Sleeman
Medical Faculty Mannheim, ECAS TRIDOMUS-Gebäude Haus C, University of Heidelberg, 68167 Mannheim, Germany; IBCS-BIP, Campus Nord, Karlsruhe Institute for Technology (KIT), 76021 Karlsruhe, Germany
Stefan Wölfl
Department of Biology, Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, 69120 Heidelberg, Germany
Matthias Philip Ebert
Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany
Christoph Meyer
Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany
Ute Hofmann
Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology and University of Tübingen, 70376 Stuttgart, Germany
Steven Dooley
Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany; Address of the Corresponding/Senior author: Department of Medicine II, Molecular Hepatology Section, Medical Faculty Mannheim, University of Heidelberg, 68167 Mannheim, Germany.
Background: The extracellular signal-regulated kinase (ERK) pathway regulates cell growth, and is hyper-activated and associated with drug resistance in hepatocellular carcinoma (HCC). Metabolic pathways are profoundly dysregulated in HCC. Whether an altered metabolic state is linked to activated ERK pathway and drug response in HCC is unaddressed. Methods: We deprived HCC cells of glutamine to induce metabolic alterations and performed various assays, including metabolomics (with 13C-glucose isotope tracing), microarray analysis, and cell proliferation assays. Glutamine-deprived cells were also treated with kinase inhibitors (e.g. Sorafenib, Erlotinib, U0126 amongst other MEK inhibitors). We performed bioinformatics analysis and stratification of HCC tumour microarrays to determine upregulated ERK gene signatures in patients. Findings: In a subset of HCC cells, the withdrawal of glutamine triggers a severe metabolic alteration and ERK phosphorylation (pERK). This is accompanied by resistance to the anti-proliferative effect of kinase inhibitors, despite pERK inhibition. High intracellular serine is a consistent feature of an altered metabolic state and contributes to pERK induction and the kinase inhibitor resistance. Blocking the ERK pathway facilitates cell proliferation by reprogramming metabolism, notably enhancing aerobic glycolysis. We have identified 24 highly expressed ERK gene signatures that their combined expression strongly indicates a dysregulated metabolic gene network in human HCC tissues. Interpretation: A severely compromised metabolism lead to ERK pathway induction, and primes some HCC cells to pro-survival phenotypes upon ERK pathway blockade. Our findings offer novel insights for understanding, predicting and overcoming drug resistance in liver cancer patients. Fund: DFG, BMBF and Sino-German Cooperation Project Keywords: Metabolic state, Kinase inhibitors, Glutamine, Serine, Proliferation, Aerobic glycolysis, HCC
