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

IF 4.3 Q1 BIOCHEMICAL RESEARCH METHODS Cell Reports Methods Pub Date : 2024-10-21 DOI:10.1016/j.crmeth.2024.100883

Frank Vrieling, Hendrik J P van der Zande, Britta Naus, Lisa Smeehuijzen, Julia I P van Heck, Bob J Ignacio, Kimberly M Bonger, Jan Van den Bossche, Sander Kersten, Rinke Stienstra

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Abstract

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.

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CENCAT 可通过生物正交非对称氨基酸标记测量蛋白质合成，从而进行免疫代谢分析。

细胞能量代谢对免疫细胞的功能起着重要作用。为了进一步推动免疫代谢研究，需要采用新方法来研究复杂样本中免疫细胞的新陈代谢。在此，我们介绍 CENCAT（通过非典型氨基酸标记的细胞能量学）。这项技术利用含炔基烃的非典型氨基酸的点击标记来测量蛋白质合成抑制，以此来替代新陈代谢活动。CENCAT 成功再现了人类原代巨噬细胞中已知的脂多糖（LPS）/干扰素（IFN）γ 和白细胞介素（IL）-4 激活的代谢特征。在外周血单核细胞中应用 CENCAT 发现，在不同激活剂的刺激下，会出现不同的代谢重新布线。最后，CENCAT 被用于分析来自不同器官的小鼠组织驻留免疫细胞的细胞代谢。根据代谢特征观察到了组织特异性聚类，这可能是由微环境引物驱动的。总之，CENCAT 提供了对免疫细胞代谢反应的宝贵见解，为研究人类和小鼠复杂样本和组织中的细胞代谢提供了一个强大的平台。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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