An integrated toolbox to profile (macrophage) immunometabolism

Sanne G.S. Verberk , Kyra de Goede (1) , Friederike Gorki (1) , Xanthe A.M.H. van Dierendonck (2) , Rafael J Argüello (3, 4, 5, 6) , Jan van den Bossche

1 VU - Vrije Universiteit Amsterdam [Amsterdam]
2 NMG - Nutrition, Metabolism and Genomics group and Nutrigenomics Consortium
3 CNRS - Centre National de la Recherche Scientifique
4 INSERM - Institut National de la Santé et de la Recherche Médicale
5 AMU - Aix Marseille Université
6 CIML - Centre d'Immunologie de Marseille - Luminy

Macrophages are plastic immune cells that can adopt several activation states. Fundamental to these functional activation states is the regulation of cellular metabolic processes. Especially in mice, the metabolic alterations underlying pro-inflammatory or homeostatic phenotypes have been assessed thoroughly using various techniques. However, researchers new to the field may encounter ambiguity in choosing which combination of techniques is best suited to profile immunometabolism. To guide readers, we provide a toolbox to assess cellular metabolism in a semi-highthroughput 96-well-plate-based format. We applied the approach to activated mouse and human macrophages by metabolically pre-screening cells, followed by measuring extracellular fluxes, mitochondrial mass and membrane potential, glucose and lipid uptake, along with the application of SCENITH. Hereby, we not only validated established activation-induced metabolic rewiring in mouse macrophages, but also uncovered new insights in human macrophage immunometabolism. By thoroughly discussing each techniques, we guide readers with a practical workflow to interrogate immunometabolism.

Mots clés: immunometabolism; macrophages; toolbox; semi-high-throughput; metabolism Motivation

Domaines: Sciences du Vivant [q-bio]

Sanne G.S. Verberk, Kyra de Goede, Friederike Gorki, Xanthe A.M.H. van Dierendonck, Rafael J Argüello, et al.. An integrated toolbox to profile (macrophage) immunometabolism. Cell Reports Methods, 2022, 2 (4), pp.100192. ⟨10.1016/j.crmeth.2022.100192⟩. ⟨hal-03856610⟩