Stable isotope metabolomics of pulmonary artery smooth muscle and endothelial cells in pulmonary hypertension and with TGF-beta treatment

Research output: Contribution to journalArticle

Authors

  • Daniel Hernandez-Saavedra
  • Linda Sanders
  • Scott Freeman
  • Julie A. Reisz
  • Michael H. Lee
  • Claudia Mickael
  • Rahul Kumar
  • Biruk Kassa
  • Sue Gu
  • Angelo D. Alessandro
  • Kurt R. Stenmark
  • Rubin M. Tuder
  • Brian B. Graham

External Institution(s)

  • University of Colorado Anschutz Medical Campus
  • University of California at San Francisco

Details

Original languageEnglish (US)
Article number413
JournalScientific reports
Volume10
Issue number1
StatusPublished - Dec 1 2020
Peer-reviewedYes

Abstract

Altered metabolism in pulmonary artery smooth muscle cells (PASMCs) and endothelial cells (PAECs) contributes to the pathology of pulmonary hypertension (PH), but changes in substrate uptake and how substrates are utilized have not been fully characterized. We hypothesized stable isotope metabolomics would identify increased glucose, glutamine and fatty acid uptake and utilization in human PASMCs and PAECs from PH versus control specimens, and that TGF-β treatment would phenocopy these metabolic changes. We used 13C-labeled glucose, glutamine or a long-chain fatty acid mixture added to cell culture media, and mass spectrometry-based metabolomics to detect and quantify 13C-labeled metabolites. We found PH PASMCs had increased glucose uptake and utilization by glycolysis and the pentose shunt, but no changes in glutamine or fatty acid uptake or utilization. Diseased PAECs had increased proximate glycolysis pathway intermediates, less pentose shunt flux, increased anaplerosis from glutamine, and decreased fatty acid β-oxidation. TGF-β treatment increased glycolysis in PASMCs, but did not recapitulate the PAEC disease phenotype. In TGF-β-treated PASMCs, glucose, glutamine and fatty acids all contributed carbons to the TCA cycle. In conclusion, PASMCs and PAECs collected from PH subjects have significant changes in metabolite uptake and utilization, partially recapitulated by TGF-β treatment.

Citation formats

APA

Hernandez-Saavedra, D., Sanders, L., Freeman, S., Reisz, J. A., Lee, M. H., Mickael, C., ... Graham, B. B. (2020). Stable isotope metabolomics of pulmonary artery smooth muscle and endothelial cells in pulmonary hypertension and with TGF-beta treatment. Scientific reports, 10(1), [413]. https://doi.org/10.1038/s41598-019-57200-5

Harvard

Hernandez-Saavedra, D, Sanders, L, Freeman, S, Reisz, JA, Lee, MH, Mickael, C, Kumar, R, Kassa, B, Gu, S, Alessandro, AD, Stenmark, KR, Tuder, RM & Graham, BB 2020, 'Stable isotope metabolomics of pulmonary artery smooth muscle and endothelial cells in pulmonary hypertension and with TGF-beta treatment', Scientific reports, vol. 10, no. 1, 413. https://doi.org/10.1038/s41598-019-57200-5