Reduced oxidative phosphorylation and increased glycolysis in human glaucoma lamina cribrosa cells

Khalid Kamel, Colm J. O'Brien, Alexander V. Zhdanov, Dmitri B. Papkovsky, Abbot F. Clark, Daniel Stamer, Mustapha Irnaten

Research output: Contribution to journalArticlepeer-review

Abstract

PURPOSE. The lamina cribrosa (LC) is a key site of damage in glaucomatous optic neuropathy. We previously found that glaucoma LC cells have an increased profibrotic gene expression, with mitochondrial dysfunction in the form of decreased mitochondrial membrane potential. Altered cell bioenergetics have recently been reported in organ fibrosis and in cancer. In this study, we carried out a systematic mitochondrial bioenergetic assessment and measured markers of alternative sources of cellular energy in normal and glaucoma LC cells. METHODS. LC cells from three glaucoma donors and three age-matched normal controls were assessed using VICTOR X4 Perkin Elmer (Waltham, MA) plate reader with different phosphorescent and luminescent probes. adenosine triphosphate levels, oxygen consumption rate, and extracellular acidification were measured and normalized to total protein content. RNA and protein expression levels of MCT1, MCT4, MTFHD2, and GLS2 were quantified using real-time RT-PCR and Western blotting. RESULTS. Glaucoma LC cells contain significantly less adenosine triphosphate (P <.05) when supplied with either glucose or galactose. They also showed significantly diminished oxygen consumption in both basal and maximal respiration with more lactic acid contribution in ECA. Both mRNA and protein expression levels of MCT1, MCT4, MTHFD2, and GLS2 were significantly increased in glaucoma LC cells. CONCLUSIONS. We demonstrate evidence of metabolic reprogramming (The Warburg effect) in glaucoma LC cells. Expression of markers of glycolysis, glutamine, and one carbon metabolism are elevated in glaucoma cells at both the mRNA and protein levels. A better understanding of bioenergetics in glaucoma may help in the development of new therapeutics.

Original languageEnglish
Article number4
JournalInvestigative Ophthalmology and Visual Science
Volume61
Issue number13
DOIs
StatePublished - 5 Nov 2020

Keywords

  • Glaucoma
  • Lamina cribrosa
  • Mitochondria
  • Warburg

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