CENCAT enables immunometabolic profiling by measuring protein synthesis via bioorthogonal noncanonical amino acid tagging.

Autor:	Vrieling F; Division of Human Nutrition and Health, Wageningen University, Wageningen, the Netherlands; Department of Internal Medicine, Radboud University Medical Center, Nijmegen, the Netherlands., van der Zande HJP; Division of Human Nutrition and Health, Wageningen University, Wageningen, the Netherlands., Naus B; Division of Human Nutrition and Health, Wageningen University, Wageningen, the Netherlands., Smeehuijzen L; Division of Human Nutrition and Health, Wageningen University, Wageningen, the Netherlands., van Heck JIP; Department of Internal Medicine, Radboud University Medical Center, Nijmegen, the Netherlands., Ignacio BJ; Department of Synthetic Organic Chemistry, Chemical Biology Lab, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, the Netherlands., Bonger KM; Department of Synthetic Organic Chemistry, Chemical Biology Lab, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, the Netherlands., Van den Bossche J; Department of Molecular Cell Biology and Immunology, Amsterdam Cardiovascular Sciences, Amsterdam Gastroenterology Endocrinology Metabolism, Amsterdam Institute for Infection and Immunity, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, the Netherlands., Kersten S; Division of Human Nutrition and Health, Wageningen University, Wageningen, the Netherlands., Stienstra R; Division of Human Nutrition and Health, Wageningen University, Wageningen, the Netherlands; Department of Internal Medicine, Radboud University Medical Center, Nijmegen, the Netherlands. Electronic address: rinke.stienstra@wur.nl.
Jazyk:	angličtina
Zdroj:	Cell reports methods [Cell Rep Methods] 2024 Oct 21; Vol. 4 (10), pp. 100883.
DOI:	10.1016/j.crmeth.2024.100883
Abstrakt:	Cellular energy metabolism significantly contributes to immune cell function. To further advance immunometabolic research, novel methods to study the metabolism of immune cells in complex samples are required. Here, we introduce CENCAT (cellular energetics through noncanonical amino acid tagging). This technique utilizes click labeling of alkyne-bearing noncanonical amino acids to measure protein synthesis inhibition as a proxy for metabolic activity. CENCAT successfully reproduced known metabolic signatures of lipopolysaccharide (LPS)/interferon (IFN)γ and interleukin (IL)-4 activation in human primary macrophages. Application of CENCAT in peripheral blood mononuclear cells revealed diverse metabolic rewiring upon stimulation with different activators. Finally, CENCAT was used to analyze the cellular metabolism of murine tissue-resident immune cells from various organs. Tissue-specific clustering was observed based on metabolic profiles, likely driven by microenvironmental priming. In conclusion, CENCAT offers valuable insights into immune cell metabolic responses, presenting a powerful platform for studying cellular metabolism in complex samples and tissues in both humans and mice. Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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