Fatty acid carbon is essential for dNTP synthesis in endothelial cells

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

doi:10.1038/nature14362

Cited by 502 publications

(695 citation statements)

References 35 publications

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“…31 GABA-induced FFAO could also be critical in endothelial cells in the tumor microenvironment, where it has been shown that FFAO is the major source for nucleotide synthesis. 32 Furthermore, GABA could be important in regulating endothelial cell permeability, as it has recently been reported that FFAO has emerged as a central regulator of endothelial cell permeability and blood vessel stability. 33 Administration of GABA markedly attenuated insulitis and systemic inflammation in mice.…”

Section: Discussionmentioning

confidence: 99%

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γ-Aminobutyric Acid Is Synthesized and Released by the Endothelium

Sen

Roy

Bandyopadhyay

et al. 2016

Circulation Research

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Rationale: Gamma aminobutyric acid (GABA), a neurotransmitter of the central nervous system, is found in the systemic circulation of humans at a concentration between 0.5 and 3 μmol/L. However, the potential source of circulating GABA and its significance on the vascular system remains unknown. We hypothesized that endothelial cells (ECs) may synthesize and release GABA to modulate some functions in the EC and after its release into the circulation.Objective: To assess whether GABA is synthesized and released by the EC and its potential functions. Methods and Results: Utilizing the human umbilical vein ECs and aortic ECs, we demonstrated for the first time that ECs synthesize and release GABA from [1-14 C]glutamate. Localization of GABA and the presence of the GABA-synthesizing enzyme, glutamic acid decarboxylase in EC were confirmed by immunostaining and immunoblot analysis, respectively. The presence of GABA was further confirmed by immunohistochemistry in the EC lining the human coronary vessel. EC-derived GABA regulated the key mechanisms of ATP synthesis, fatty acid, and pyruvate oxidation in EC. GABA protected EC by inhibiting the reactive oxygen species generation and prevented monocyte adhesion by attenuating vascular cell adhesion molecule -1 and monocyte chemoattractant protein-1 expressions. GABA had no relaxing effect on rat aortic rings. GABA exhibited a dose-dependent fall in blood pressure. However, the fall in BP was abolished after pretreatment with pentolinium. from Perkin Elmer, MA, catalog NEC290E050UC, NEC255050UC, and NEC559010UC. All other chemicals unless specified were obtained from Sigma. Conclusions: Cell CultureHuman umbilical vein endothelial cells (HUVECs) were freshly isolated from umbilical cords from different donors after normal delivery (with approval from the Institutional Review Board of University of California at Los Angeles, LA, CA). Primary cultures of HUVEC were prepared as described 14 and used at passages 1 or 2. Primary cultures of human aortic endothelial cells (HAEC) at passage 2 or 3 were a generous gift from Dr M. Navab in the Department of Medicine, University of California, Los Angeles, 90095-1732, USA. Both HAEC and HUVEC were cultured in endothelial cell basal medium-2 (Lonza), and the medium was supplemented with Endothelial Growth Medium-2 Bullet kit (CC3162; Lonza) as per the manufacturer's instructions as described previously. 15PC12 cells were a generous gift from Dr E. Schweitzer, Semel Institute of Neuroscience and Human Behavior, University of California, Los Angeles. PC12 cells were cultured in DMEM supplemented with 5% FCS and 5% Horse serum. Human skin fibroblast (HSF) cells were a gift from Dr H. Coller, Molecular Cell and Developmental Biology, University of California, Los Angeles. HSF and THP1 were cultured in DMEM supplemented with 10% FBS. All cells were cultured until 70% confluent before downstream manipulations. Cell culture treatments were done under sterile conditions, for 24 hours unless specified. Glutamic acid decarboxylase (GAD) doubl...

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Section: Discussionmentioning

confidence: 99%

“…[31][32][33] Therefore, we assessed whether GABA promoted fatty acid oxidation in HUVEC. As fatty acid oxidation is critically regulated at the level of its mitochondrial entry, we investigated whether GABA modulated both uptake and subsequent oxidation of fatty acids in HUVEC.…”

Section: Gaba Promoted Mitochondrial Free Fatty Acid Uptake and Oxidamentioning

confidence: 99%

γ-Aminobutyric Acid Is Synthesized and Released by the Endothelium

Sen

Roy

Bandyopadhyay

et al. 2016

Circulation Research

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“…Light also could inhibit de novo purine synthesis by decreasing the amount of aspartate in the cell. Recent studies showed that decreases in nucleotide levels correlate with reduced synthesis of aspartate (52)(53)(54)(55)(56). These are reasonable mechanisms by which light could regulate de novo purine synthesis.…”

Section: Illumination Influences Nucleotide Metabolismmentioning

confidence: 99%

Phototransduction Influences Metabolic Flux and Nucleotide Metabolism in Mouse Retina

Rountree

Cleghorn³

et al. 2016

Journal of Biological Chemistry

100

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Production of energy in a cell must keep pace with demand.Photoreceptors use ATP to maintain ion gradients in darkness, whereas in light they use it to support phototransduction. Matching production with consumption can be accomplished by coupling production directly to consumption. Alternatively, production can be set by a signal that anticipates demand. In this report we investigate the hypothesis that signaling through phototransduction controls production of energy in mouse retinas. We found that respiration in mouse retinas is not coupled tightly to ATP consumption. By analyzing metabolic flux in mouse retinas, we also found that phototransduction slows metabolic flux through glycolysis and through intermediates of the citric acid cycle. We also evaluated the relative contributions of regulation of the activities of ␣-ketoglutarate dehydrogenase and the aspartate-glutamate carrier 1. In addition, a comprehensive analysis of the retinal metabolome showed that phototransduction also influences steady-state concentrations of 5-GMP, ribose-5-phosphate, ketone bodies, and purines. Warburg et al.(1) and Krebs (2) reported in the 1920s that tumors and retinas rely on aerobic glycolysis. Some of the biochemical mechanisms by which cancer cells adapt to aerobic glycolysis have been gleaned from investigations of specific metabolic adaptations of cancer cells either in culture or in a tumor (3, 4). Retinas offer distinct advantages for investigating aerobic glycolysis. They have high metabolic rates, a uniquely laminated structure, and the primary signaling pathway by which retinas respond to light is defined clearly.Retinas convert 80 -96% of glucose they consume into lactic acid (5-9), similar to the extent of aerobic glycolysis that fuels cancer cells (3). Aerobic glycolysis occurs primarily in photoreceptors (10) where the energy demands are very different in darkness than in light (5,7,(11)(12)(13). In darkness energy is consumed within the inner segments to support ion pumping (5, 13). In light energy is consumed by the outer segments (OS) 2 to support phototransduction and regeneration of visual pigments. A photoreceptor neuron also performs anabolic metabolism to replace the ϳ10% of its OS material that is lost each day to phagocytosis by the retinal pigmented epithelium (14, 15).Some of the carbons in glucose consumed by a retina reach the mitochondrial matrix where they are oxidized in biochemical reactions that reduce NAD ϩ to NADH. Transfer of electrons from NADH to O 2 then generates a proton gradient across the mitochondrial inner membrane. In some tissues dissipation of the proton gradient is coupled tightly to ATP demand. In others, proton leakage can dissipate the gradient even without ATP synthesis (16). In this report we show that mitochondria in retinas are more uncoupled than mitochondria in other tissues.The ability of a photoreceptor to respond to light, to release neurotransmitter, to regenerate visual pigment, to renew itself, and to remain viable requires that production of energy keeps pace...

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“…54 Unexpectedly, ECs do not rely on fatty acid oxidation for the production of ATP or redox homeostasis, but use fatty acids as a carbon source for the synthesis of DNA during cell replication, unlike multiple other cell types. 55 Additional metabolic pathways, including the pentose phosphate pathway, amino acid metabolism and others have been poorly studied in ECs in relation to (pathological) angiogenesis. 52,56 A relationship between metabolism and the cell cycle in ECs has not been studied at all.…”

Section: Endothelial Cell Metabolismmentioning

confidence: 99%

Metabolic control of the cell cycle

et al. 2015

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# These authors contributed equally to this work. C ell division is a metabolically demanding process, requiring the production of large amounts of energy and biomass. Not surprisingly therefore, a cell's decision to initiate division is codetermined by its metabolic status and the availability of nutrients. Emerging evidence reveals that metabolism is not only undergoing substantial changes during the cell cycle, but it is becoming equally clear that metabolism regulates cell cycle progression. Here, we overview the emerging role of those metabolic pathways that have been best characterized to change during or influence cell cycle progression. We then studied how Notch signaling, a key angiogenic pathway that inhibits endothelial cell (EC) proliferation, controls EC metabolism (glycolysis) during the cell cycle.

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Fatty acid carbon is essential for dNTP synthesis in endothelial cells

Cited by 502 publications

References 35 publications

γ-Aminobutyric Acid Is Synthesized and Released by the Endothelium

γ-Aminobutyric Acid Is Synthesized and Released by the Endothelium

Phototransduction Influences Metabolic Flux and Nucleotide Metabolism in Mouse Retina

Metabolic control of the cell cycle

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