Bioorthogonal tools to study fatty acid uptake in immune cells

Bertheussen, K.

The cellular metabolism of immune cells constitutes an important part of their proper function and activation. Using T cells as an example, this phenomenon is highlighted by the metabolic reprogramming that occurs in these cells upon activation. The metabolic reprogramming is supported by an increased uptake of exogenous nutrients, such as glucose, amino acids, and fatty acids (FAs). The ability to unravel this complex interface between the immune system and cellular metabolism relies on the development of appropriate chemical tools. The aim of this thesis was to develop sterculic acid (StA) as a bioorthogonal analogue of the immunomodulatory FA oleic acid (OA), and to use this as a tool to study the uptake of StA in different immune cells. Here, we report the first use of StA as a bioorthogonal analogue of OA. It was shown that StA was readily taken up in vitro by a plethora of immune cells and could react with tetrazine-modified reporter molecules (e.g. fluorophores) via an...Show moreThe cellular metabolism of immune cells constitutes an important part of their proper function and activation. Using T cells as an example, this phenomenon is highlighted by the metabolic reprogramming that occurs in these cells upon activation. The metabolic reprogramming is supported by an increased uptake of exogenous nutrients, such as glucose, amino acids, and fatty acids (FAs). The ability to unravel this complex interface between the immune system and cellular metabolism relies on the development of appropriate chemical tools. The aim of this thesis was to develop sterculic acid (StA) as a bioorthogonal analogue of the immunomodulatory FA oleic acid (OA), and to use this as a tool to study the uptake of StA in different immune cells. Here, we report the first use of StA as a bioorthogonal analogue of OA. It was shown that StA was readily taken up in vitro by a plethora of immune cells and could react with tetrazine-modified reporter molecules (e.g. fluorophores) via an inverse electron-demand Diels-Alder reaction, presenting a non-toxic, live-cell compatible alternative to other commonly used bioorthogonal reactions. The versatility of this approach allowed for multiplexing with other bioorthogonal reactions, permitting the simultaneous study of multiple biomolecules (Chapter 2). The workflow could further be adapted and applied to study the lipidation of proteins as a PTM (Chapter 3), as well as to investigate phenotypic and multiomic differences between T cells with a differential StA-uptake (Chapter 4). The adaptability of the workflow emphasises its strength and applicability as a chemical tool to study FA uptake.
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Bioorthogonal tools to study fatty acid uptake in immune cells