| Autor: |
Mavin E; Institute of Cellular Medicine, Newcastle University, Newcastle Upon Tyne, United Kingdom., Verdon B; Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, United Kingdom., Carrie S; Institute of Health and Society, Newcastle University, Newcastle Upon Tyne, United Kingdom., Saint-Criq V; Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, United Kingdom., Powell J; Institute of Cellular Medicine, Newcastle University, Newcastle Upon Tyne, United Kingdom., Kuttruff CA; Medicinal Chemistry, Boehringer Ingelheim Pharma, Biberach an der Riss, Germany., Ward C; Institute of Cellular Medicine, Newcastle University, Newcastle Upon Tyne, United Kingdom.; Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, United Kingdom., Garnett JP; Institute of Cellular Medicine, Newcastle University, Newcastle Upon Tyne, United Kingdom.; Immunology and Respiratory Diseases Research, Boehringer Ingelheim Pharma, Biberach an der Riss, Germany., Miwa S; Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle Upon Tyne, United Kingdom. |
| Abstrakt: |
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. |
