Tuesday, January 23rd, Utah, USA. Poster #2077 "Modelling tumour hypoxia through parallel analysis of cellular oxygenation, glycolysis flux and mitochondrial function."
In conjunction with the MetaCell Project, researchers from the University of Oxford will be presenting a poster at the Keystone Symposia Meeting on Tumor Metabolism, a premier conference for tumor metabolism experts to discuss emerging concepts and new research in the regulation and role of cancer metabolism in tumor growth. The poster is entitled: “Modelling tumor hypoxia through parallel analysis of cellular oxygenation, glycolysis flux and mitochondrial function” and will be presented on Tuesday, January 23rd during Poster Session 2 (Poster #2077).
In the poster, a methodology is described for modeling tumor hypoxia in vitro using the MitoXpress® Intra Intracellular Oxygen Assay probe measured on a fluorescence plate reader equipped with an atmospheric control unit (CLARIOstar®, BMG Labtech). The data presented invalidates the assumption that cellular oxygenation can be inferred from ambient oxygen measurements and illustrates how this assumption can lead to erroneous conclusions regarding the relationship between oxygen concentration, HIF stabilization and related metabolic adaptations.
Modelling tumour hypoxia through parallel analysis of cellular oxygenation, glycolysis flux and mitochondrial function.
M. Potter1, M. Schwalfenberg2, C. Yalaz1, R. McGarrigle2, C. Zois1, A. Harris1, J. Hynes2, K Morten1
1University of Oxford, Oxford, UK, 2Luxcel Biosciences, Cork, Ireland
Malignant transformation is associated with significant metabolic reprogramming, as cancer cells strive to maintain ATP supply and fuel catabolism, often achieved by increasing glycolytic flux. A key factor influencing this metabolic programming is oxygen availability. This is particularly true of solid tumours, where oncogene-driven proliferation causes nutrient and oxygen deprivation, aberrant angiogenesis, and the activation of such O2-sensitive survival pathways. Delineating the relationship between oxygenation and such metabolic reprogramming is therefore key to a deeper understanding of these processes and to the development of more effective therapeutic interventions. Detailed in vitro analysis has however been limited due to the absence of methodologies capable of controlling and monitoring cellular oxygenation, while in parallel, probing other key parameters such as glycolytic flux and HIF stabilisation. Here we describe a methodology for modelling tumour hypoxia in vitro using a fluorescence plate reader equipped with an atmospheric control unit (CLARIOstar) in combination with a nanoparticulate intracellular oxygen probe (MitoXpress-Intra). Data is presented illustrating that the depth of hypoxia experienced by the cell model is impacted significantly by respiratory activity, and that this additional oxygen deprivation is a dynamic process, effected by respiratory substrate availability and related metabolic poise. Multiplexing with the pH-sensitive probe (pH-Xtra™) allows real-time monitoring of cellular oxygenation and glycolytic activity allowing the interrelationship between oxygenation and glycolytic flux to be characterised. Together these data invalidate the assumption that cellular oxygenation can be inferred from ambient oxygen measurements and if ignored, the significant and dynamic deviations between ambient O2 and oxygenation at the cellular level lead to erroneous conclusions regarding the relationship between oxygen concentration, HIF stabilisation and related metabolic adaptations. This, in turn, can impair the physiological relevance of experimental observations.