Real-time measurement of cellular bioenergetics in fully differentiated human nasal epithelial cells grown at air-liquid-interface
| Autor: | Sean Carrie, Christian A. Kuttruff, Chris Ward, Satomi Miwa, James P. Garnett, Jason Powell, Emily Mavin, Vinciane Saint-Criq, Bernard Verdon |
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| Přispěvatelé: | Institute for Cell and Molecular Biosciences, Newcastle University [Newcastle] |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Pulmonary and Respiratory Medicine Physiology [SDV]Life Sciences [q-bio] Biology Nose 03 medical and health sciences 0302 clinical medicine Metabolomics Computer Systems Physiology (medical) medicine Humans Glycolysis Respiratory system Cells Cultured L-Lactate Dehydrogenase Air Respiratory disease Cell Differentiation Epithelial Cells airway epithelium Cell Biology medicine.disease Seahorse Epithelium Cell biology 030104 developmental biology medicine.anatomical_structure Glucose Cell culture Anaerobic glycolysis 030220 oncology & carcinogenesis Innovative Methodology Respiratory epithelium Energy Metabolism Acids metabolism |
| Zdroj: | American Journal of Physiology-Lung Cellular and Molecular Physiology American Journal of Physiology-Lung Cellular and Molecular Physiology, American Physiological Society, 2020, 318 (6), pp.L1158-L1164. ⟨10.1152/ajplung.00414.2019⟩ |
| ISSN: | 1040-0605 1522-1504 |
| DOI: | 10.1152/ajplung.00414.2019⟩ |
| Popis: | Shifts in cellular metabolic phenotypes have the potential to cause disease-driving processes in respiratory disease. The respiratory epithelium is particularly susceptible to metabolic shifts in disease, but our understanding of these processes is limited by the incompatibility of the technology required to measure metabolism in real-time with the cell culture platforms used to generate differentiated respiratory epithelial cell types. Thus, to date, our understanding of respiratory epithelial metabolism has been restricted to that of basal epithelial cells in submerged culture, or via indirect end point metabolomics readouts in lung tissue. Here we present a novel methodology using the widely available Seahorse Analyzer platform to monitor real-time changes in the cellular metabolism of fully differentiated primary human airway epithelial cells grown at air-liquid interface (ALI). We show increased glycolytic, but not mitochondrial, ATP production rates in response to physiologically relevant increases in glucose availability. We also show that pharmacological inhibition of lactate dehydrogenase is able to reduce glucose-induced shifts toward aerobic glycolysis. This method is timely given the recent advances in our understanding of new respiratory epithelial subtypes that can only be observed in vitro through culture at ALI and will open new avenues to measure real-time metabolic changes in healthy and diseased respiratory epithelium, and in turn the potential for the development of novel therapeutics targeting metabolic-driven disease phenotypes. |
| Databáze: | OpenAIRE |
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