[en] Modeling tumor metabolism in vitro remains challenging. Here, we used galactose as an in vitro tool compound to mimic glycolytic limitation. In contrast to the established idea that high glycolytic flux reduces pyruvate kinase isozyme M2 (PKM2) activity to support anabolic processes, we have discovered that glycolytic limitation also affects PKM2 activity. Surprisingly, despite limited carbon availability and energetic stress, cells induce a near-complete block of PKM2 to divert carbons toward serine metabolism. Simultaneously, TCA cycle flux is sustained, and oxygen consumption is increased, supported by glutamine. Glutamine not only supports TCA cycle flux but also serine synthesis via distinct mechanisms that are directed through PKM2 inhibition. Finally, deleting mitochondrial one-carbon (1C) cycle reversed the PKM2 block, suggesting a potential formate-dependent crosstalk that coordinates mitochondrial 1C flux and cytosolic glycolysis to support cell survival and proliferation during nutrient-scarce conditions.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
BENZARTI, Mohaned ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM)
NEISES, Laura ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine > Molecular and Functional Neurobiology > Team Anne GRÜNEWALD ; Cancer Metabolism Group, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
Oudin, Anais; NORLUX Neuro-Oncology Laboratory, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
Krötz, Christina; Cancer Metabolism Group, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
VIRY, Elodie ; University of Luxembourg > Faculty of Science, Technology and Medicine > Department of Life Sciences and Medicine > Team Serge HAAN ; Tumor Stroma Interactions, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
Gargiulo, Ernesto; Tumor Stroma Interactions, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
Pulido, Coralie; Animal Facility, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
SCHMOETTEN, Maryse ; University of Luxembourg > Faculty of Science, Technology and Medicine > Department of Life Sciences and Medicine > Team Elisabeth LETELLIER
POZDEEV, Vitaly ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM)
Lorenz, Nadia I; Dr. Senckenberg Institute of Neurooncology, University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany, German Cancer Consortium, Partner Site Frankfurt/Mainz, Frankfurt am Main, Germany, Frankfurt Cancer Institute (FCI), University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany, University Cancer Center Frankfurt (UCT), University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany
Ronellenfitsch, Michael W; Dr. Senckenberg Institute of Neurooncology, University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany, German Cancer Consortium, Partner Site Frankfurt/Mainz, Frankfurt am Main, Germany, Frankfurt Cancer Institute (FCI), University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany, University Cancer Center Frankfurt (UCT), University Hospital Frankfurt, Goethe University, Frankfurt am Main, Germany
Sumpton, David; Cancer Research U.K. Scotland Institute, Switchback Road, Bearsden, Glasgow G61 1BD, UK
WARMOES, Marc Omer ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine > Scientific Central Services > Metabolomics Platform
Jaeger, Christian; Metabolomics Platform, Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Belvaux, Luxembourg
Lesur, Antoine; Cancer Metabolism Group, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
Becker, Björn; Cancer Metabolism Group, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
MOUSSAY, Etienne ; University of Luxembourg ; Tumor Stroma Interactions, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
PAGGETTI, Jerome ; University of Luxembourg ; Tumor Stroma Interactions, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg
NICLOU, Simone P. ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM)
LETELLIER, Elisabeth ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM)
MEISER, Johannes ; University of Luxembourg ; Cancer Metabolism Group, Department of Cancer Research, Luxembourg Institute of Health, Luxembourg, Luxembourg. Electronic address: johannes.meiser@lih.lu
M.B. is supported by the Fondation du Pélican de Mie et Pierre Hippert-Faber , under the aegis of the Fondation de Luxembourg . M.B. and J.M. are supported by the Luxembourg National Research Fund (FNR) ATTRACT program ( A18/BM/11809970 ). M.B., J.M., and E.L. are supported by FNR-CORE grant ( C21/BM/15718879/1cFlex ). E.L. is supported by the FNR-CORE program ( C16/BM/11282028 and C20/BM/14591557 ), by a Proof of Concept FNR grant ( PoC/18/12554295 ), a PRIDE17/11823097 , and by i2Tron ( PRIDE19/14254520 ). E.G., J.P., and E.M. are supported by grants from the FNR and Fondation Cancer ( PRIDE15/10675146/CANBIO , C20/BM/14582635 , and C20/BM/14592342 ). E.V. is supported by FNRS-Télévie ( 7.4509.20 and 7.4572.22 ).We are grateful to Stephanie Kreis (University of Luxembourg) for providing us with B16-F10, A375, and SKMeL28 cells; Clement Thomas (LIH, Luxembourg) for providing 4T1 and BT-20 cells; Lewis Cantley (Harvard Medical School) for providing MDA-MB-468 WT and MTHFD1LKO cells; and Dimitar Efremov (ICGEB, Trieste) for providing TCL1-355 cells. We are grateful for all the technical and analytical support from the different metabolomics platforms toward the work presented here and, in specific, Francois Bernardin from the metabolomics platform at LIH, and Xiangyi Dong, Floriane Gavotto, and Lucia Gallucci from the LCSB Metabolomics Platform (RRID:SCR 024769). We are thankful for the support of the National Cytometry Platform (Quantitative Biology Unit, LIH) for support with flow cytometry measurements and analysis. Figures 1A, 1B, 3K, 5A, 5H, 6A and 7A were created with BioRender.com. M.B. is supported by the Fondation du Pélican de Mie et Pierre Hippert-Faber, under the aegis of the Fondation de Luxembourg. M.B. and J.M. are supported by the Luxembourg National Research Fund (FNR) ATTRACT program (A18/BM/11809970). M.B. J.M. and E.L. are supported by FNR-CORE grant (C21/BM/15718879/1cFlex). E.L. is supported by the FNR-CORE program (C16/BM/11282028 and C20/BM/14591557), by a Proof of Concept FNR grant (PoC/18/12554295), a PRIDE17/11823097, and by i2Tron (PRIDE19/14254520). E.G. J.P. and E.M. are supported by grants from the FNR and Fondation Cancer (PRIDE15/10675146/CANBIO, C20/BM/14582635, and C20/BM/14592342). E.V. is supported by FNRS-Télévie (7.4509.20 and 7.4572.22). M.B. and J.M. conceptualized and designed the study. M.B. L.N. C.K. and B.B. performed all of the in vitro experiments. A.O. E.V. E.G. V.P. M.S. C.P. J.P. E.M. S.P.N. and E.L. assisted in the design and performing of the in vivo work. L.N. N.I.L. and M.W.R. aided in the generation of different KO cell lines used. D.S. M.W. C.J. and A.L. performed YSI, LC-MS, GC-MS, and IC-MS measurements, while M.B. performed the corresponding data analysis according to instructions from D.S. M.W. C.J. A.L. and J.M. Original draft was written by M.B. and J.M. Review and editing was done by all authors. The authors of the presented study declare no competing interests.
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