3-hydroxypyruvate destabilizes hypoxia inducible factor and induces angiostasis

Charandeep Singh, Amit Sharma, George B. Hoppe, Weilin Song, Youstina Bolok, Jonathan E Sears

Research output: Contribution to journalArticle

Abstract

PURPOSE. Transcriptional analysis of retina protected by hypoxia-inducible factor (HIF) stabilization demonstrates an increase in genes associated with aerobic glycolysis. We hypothesized that since protection is associated with a change in metabolism, oxygen-induced metabolites might transduce oxygen toxicity. We used global metabolic profiling to identify retinal metabolites increased in hyperoxia compared to normoxia. METHODS. Untargeted gas chromatography mass spectroscopy (GC-MS) was performed on both mouse retina samples collected in hyperoxia and on primary human retinal endothelial cells, each with and without HIF stabilization. After identifying 3-hydropxypyruvate (3OH-pyruvate) as a unique hyperoxic metabolite, endothelial cells in culture and choroidal explants were challenged with 3OH-pyruvate in order to determine how this glycolytic intermediate was metabolized, and whether it had an effect on angiogenesis. RESULTS. 3OH-pyruvate was one of five metabolites at least 2.0-fold elevated in hyperoxia with a P value < 0.1. Once metabolized by endothelial cells, 3OH-pyruvate led to a 20-fold increase in 3-phosphoglycerate and a 4-fold increase in serine when cells were treated with Roxadustat to induce HIF stabilization. 3OH-pyruvate, but not pyruvate, destabilized HIF in endothelial cells with an increase in proline hydroxylation. 3OH-pyruvate was angiostatic in choroidal explant assays. CONCLUSIONS. 3OH-pyruvate is a unique metabolite induced by hyperoxia that destabilizes HIF at least in part by a canonical pathway. 3OH-pyruvate induces angiostasis in vitro. HIF stabilization increases serine biosynthesis in vitro and in vivo.

LanguageEnglish (US)
Pages3440-3448
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume59
Issue number8
DOIs
StatePublished - Jul 1 2018
Externally publishedYes

Fingerprint

Pyruvic Acid
Hyperoxia
Endothelial Cells
Serine
Retina
Oxygen
Hypoxia
Glycolysis
Hydroxylation
Proline
Gas Chromatography
Mass Spectrometry
Cell Culture Techniques
Genes

Keywords

  • 3-hydroxypyruvate
  • Hypoxia inducible factor
  • Retinal metabolism
  • Retinopathy of prematurity
  • Serine

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

3-hydroxypyruvate destabilizes hypoxia inducible factor and induces angiostasis. / Singh, Charandeep; Sharma, Amit; Hoppe, George B.; Song, Weilin; Bolok, Youstina; Sears, Jonathan E.

In: Investigative Ophthalmology and Visual Science, Vol. 59, No. 8, 01.07.2018, p. 3440-3448.

Research output: Contribution to journalArticle

Singh, Charandeep ; Sharma, Amit ; Hoppe, George B. ; Song, Weilin ; Bolok, Youstina ; Sears, Jonathan E. / 3-hydroxypyruvate destabilizes hypoxia inducible factor and induces angiostasis. In: Investigative Ophthalmology and Visual Science. 2018 ; Vol. 59, No. 8. pp. 3440-3448.
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T1 - 3-hydroxypyruvate destabilizes hypoxia inducible factor and induces angiostasis

AU - Singh, Charandeep

AU - Sharma, Amit

AU - Hoppe, George B.

AU - Song, Weilin

AU - Bolok, Youstina

AU - Sears, Jonathan E

PY - 2018/7/1

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N2 - PURPOSE. Transcriptional analysis of retina protected by hypoxia-inducible factor (HIF) stabilization demonstrates an increase in genes associated with aerobic glycolysis. We hypothesized that since protection is associated with a change in metabolism, oxygen-induced metabolites might transduce oxygen toxicity. We used global metabolic profiling to identify retinal metabolites increased in hyperoxia compared to normoxia. METHODS. Untargeted gas chromatography mass spectroscopy (GC-MS) was performed on both mouse retina samples collected in hyperoxia and on primary human retinal endothelial cells, each with and without HIF stabilization. After identifying 3-hydropxypyruvate (3OH-pyruvate) as a unique hyperoxic metabolite, endothelial cells in culture and choroidal explants were challenged with 3OH-pyruvate in order to determine how this glycolytic intermediate was metabolized, and whether it had an effect on angiogenesis. RESULTS. 3OH-pyruvate was one of five metabolites at least 2.0-fold elevated in hyperoxia with a P value < 0.1. Once metabolized by endothelial cells, 3OH-pyruvate led to a 20-fold increase in 3-phosphoglycerate and a 4-fold increase in serine when cells were treated with Roxadustat to induce HIF stabilization. 3OH-pyruvate, but not pyruvate, destabilized HIF in endothelial cells with an increase in proline hydroxylation. 3OH-pyruvate was angiostatic in choroidal explant assays. CONCLUSIONS. 3OH-pyruvate is a unique metabolite induced by hyperoxia that destabilizes HIF at least in part by a canonical pathway. 3OH-pyruvate induces angiostasis in vitro. HIF stabilization increases serine biosynthesis in vitro and in vivo.

AB - PURPOSE. Transcriptional analysis of retina protected by hypoxia-inducible factor (HIF) stabilization demonstrates an increase in genes associated with aerobic glycolysis. We hypothesized that since protection is associated with a change in metabolism, oxygen-induced metabolites might transduce oxygen toxicity. We used global metabolic profiling to identify retinal metabolites increased in hyperoxia compared to normoxia. METHODS. Untargeted gas chromatography mass spectroscopy (GC-MS) was performed on both mouse retina samples collected in hyperoxia and on primary human retinal endothelial cells, each with and without HIF stabilization. After identifying 3-hydropxypyruvate (3OH-pyruvate) as a unique hyperoxic metabolite, endothelial cells in culture and choroidal explants were challenged with 3OH-pyruvate in order to determine how this glycolytic intermediate was metabolized, and whether it had an effect on angiogenesis. RESULTS. 3OH-pyruvate was one of five metabolites at least 2.0-fold elevated in hyperoxia with a P value < 0.1. Once metabolized by endothelial cells, 3OH-pyruvate led to a 20-fold increase in 3-phosphoglycerate and a 4-fold increase in serine when cells were treated with Roxadustat to induce HIF stabilization. 3OH-pyruvate, but not pyruvate, destabilized HIF in endothelial cells with an increase in proline hydroxylation. 3OH-pyruvate was angiostatic in choroidal explant assays. CONCLUSIONS. 3OH-pyruvate is a unique metabolite induced by hyperoxia that destabilizes HIF at least in part by a canonical pathway. 3OH-pyruvate induces angiostasis in vitro. HIF stabilization increases serine biosynthesis in vitro and in vivo.

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KW - Retinopathy of prematurity

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