Fatty acid carbon is essential for dNTP synthesis in endothelial cells

Sandra Schoors; Ulrike Bruning; Rindert Missiaen; Karla Cs Queiroz; Gitte Borgers; Ilaria Elia; Annalisa Zecchin; Anna Rita Cantelmo; Stefan Christen; Jermaine Goveia; Ward Heggermont; Lucica Goddé; Stefan Vinckier; Paul P Van Veldhoven; Guy Eelen; Luc Schoonjans; Holger Gerhardt; Mieke Dewerchin; Myriam Baes; Katrien De Bock; Bart Ghesquière; Sophia Y Lunt; Sarah-Maria Fendt; Peter Carmeliet

doi:10.1038/nature14362

Fatty acid carbon is essential for dNTP synthesis in endothelial cells

Nature. 2015 Apr 9;520(7546):192-197. doi: 10.1038/nature14362. Epub 2015 Apr 1.

Authors

Sandra Schoors^#^{1

2}, Ulrike Bruning^#^{1

2}, Rindert Missiaen^{1

2}, Karla Cs Queiroz^{1

2}, Gitte Borgers^{1

2}, Ilaria Elia^{3

4}, Annalisa Zecchin^{1

2}, Anna Rita Cantelmo^{1

2}, Stefan Christen^{3

4}, Jermaine Goveia^{1

2}, Ward Heggermont⁵, Lucica Goddé^{1

2}, Stefan Vinckier^{1

2}, Paul P Van Veldhoven⁶, Guy Eelen^{1

2}, Luc Schoonjans^{1

2}, Holger Gerhardt^{7

8

9}, Mieke Dewerchin^{1

2}, Myriam Baes¹⁰, Katrien De Bock^{1

2

11}, Bart Ghesquière^{1

2}, Sophia Y Lunt¹², Sarah-Maria Fendt^{3

4}, Peter Carmeliet^{1

2}

Affiliations

¹ Laboratory of Angiogenesis and Neurovascular link, Department of Oncology, University of Leuven, Leuven, B-3000, Belgium.
² Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center, VIB, Leuven, B-3000, Belgium.
³ Laboratory of Cellular Metabolism and Metabolic Regulation, Department of Oncology, University of Leuven, Leuven, B-3000, Belgium.
⁴ Laboratory of Cellular Metabolism and Metabolic Regulation, Vesalius Research Center, VIB, Leuven, B-3000, Belgium.
⁵ Center for Molecular & Vascular Biology, KU Leuven; Division of Clinical Cardiology, UZ Leuven, B-3000, Belgium.
⁶ Laboratory of Lipid biochemistry and protein interactions, University of Leuven, B-3000, Leuven, Belgium.
⁷ Vascular Patterning Laboratory, Department of Oncology, University of Leuven, Leuven, B-3000, Belgium.
⁸ Vascular Patterning Laboratory, Vesalius Research Center, VIB, Leuven, B-3000, Belgium.
⁹ Integrative Vascular Biology Laboratory, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
¹⁰ Laboratory of Cell Metabolism, Department of pharmaceutical and pharmacological sciences, University of Leuven, Leuven, B-3000,Belgium.
¹¹ Exercise Physiology Research Group, Department of Kinesiology, University of Leuven, Leuven, B-3001, Belgium.
¹² Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan, USA.

^# Contributed equally.

Abstract

The metabolism of endothelial cells during vessel sprouting remains poorly studied. Here we report that endothelial loss of CPT1A, a rate-limiting enzyme of fatty acid oxidation (FAO), causes vascular sprouting defects due to impaired proliferation, not migration, of human and murine endothelial cells. Reduction of FAO in endothelial cells did not cause energy depletion or disturb redox homeostasis, but impaired de novo nucleotide synthesis for DNA replication. Isotope labelling studies in control endothelial cells showed that fatty acid carbons substantially replenished the Krebs cycle, and were incorporated into aspartate (a nucleotide precursor), uridine monophosphate (a precursor of pyrimidine nucleoside triphosphates) and DNA. CPT1A silencing reduced these processes and depleted endothelial cell stores of aspartate and deoxyribonucleoside triphosphates. Acetate (metabolized to acetyl-CoA, thereby substituting for the depleted FAO-derived acetyl-CoA) or a nucleoside mix rescued the phenotype of CPT1A-silenced endothelial cells. Finally, CPT1 blockade inhibited pathological ocular angiogenesis in mice, suggesting a novel strategy for blocking angiogenesis.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Acetic Acid / pharmacology
Adenosine Triphosphate / metabolism
Animals
Blood Vessels / cytology
Blood Vessels / drug effects
Blood Vessels / metabolism
Blood Vessels / pathology
Carbon / metabolism*
Carnitine O-Palmitoyltransferase / antagonists & inhibitors
Carnitine O-Palmitoyltransferase / deficiency
Carnitine O-Palmitoyltransferase / genetics
Carnitine O-Palmitoyltransferase / metabolism
Cell Line, Tumor
Cell Proliferation / drug effects
Citric Acid Cycle
DNA / biosynthesis
Disease Models, Animal
Endothelial Cells / cytology
Endothelial Cells / drug effects
Endothelial Cells / enzymology
Endothelial Cells / metabolism*
Fatty Acids / chemistry*
Fatty Acids / metabolism*
Gene Silencing
Glucose / metabolism
Human Umbilical Vein Endothelial Cells / cytology
Human Umbilical Vein Endothelial Cells / drug effects
Human Umbilical Vein Endothelial Cells / metabolism
Human Umbilical Vein Endothelial Cells / pathology
Humans
Mice
Neovascularization, Pathologic / drug therapy
Neovascularization, Pathologic / metabolism
Neovascularization, Pathologic / pathology
Nucleotides / biosynthesis*
Nucleotides / chemistry
Nucleotides / pharmacology
Oxidation-Reduction / drug effects
Retinopathy of Prematurity / drug therapy
Retinopathy of Prematurity / metabolism
Retinopathy of Prematurity / pathology

Substances

Fatty Acids
Nucleotides
Carbon
Adenosine Triphosphate
DNA
Carnitine O-Palmitoyltransferase
Glucose
Acetic Acid

Grants and funding

269073/ERC_/European Research Council/International