Cold exposure induces dynamic changes in circulating triacylglycerol species, which is dependent on intracellular lipolysis: A randomized cross-over trial

Maaike E Straat, Lucas Jurado-Fasoli, Zhixiong Ying, Kimberly J Nahon, Laura G M Janssen, Mariëtte R Boon, Gernot F Grabner, Sander Kooijman, Robert Zimmermann, Martin Giera, Patrick C N Rensen, Borja Martinez-Tellez, Maaike E Straat, Lucas Jurado-Fasoli, Zhixiong Ying, Kimberly J Nahon, Laura G M Janssen, Mariëtte R Boon, Gernot F Grabner, Sander Kooijman, Robert Zimmermann, Martin Giera, Patrick C N Rensen, Borja Martinez-Tellez

Abstract

Background: The application of cold exposure has emerged as an approach to enhance whole-body lipid catabolism. The global effect of cold exposure on the lipidome in humans has been reported with mixed results depending on intensity and duration of cold.

Methods: This secondary study was based on data from a previous randomized cross-over trial (ClinicalTrials.gov ID: NCT03012113). We performed sequential lipidomic profiling in serum during 120 min cold exposure of human volunteers. Next, the intracellular lipolysis was blocked in mice (eighteen 10-week-old male wild-type mice C57BL/6J) using a small-molecule inhibitor of adipose triglyceride lipase (ATGL; Atglistatin), and mice were exposed to cold for a similar duration. The quantitative lipidomic profiling was assessed in-depth using the Lipidyzer platform.

Findings: In humans, cold exposure gradually increased circulating free fatty acids reaching a maximum at 60 min, and transiently decreased total triacylglycerols (TAGs) only at 30 min. A broad range of TAG species was initially decreased, in particular unsaturated and polyunsaturated TAG species with ≤5 double bonds, while after 120 min a significant increase was observed for polyunsaturated TAG species with ≥6 double bonds in humans. The mechanistic study in mice revealed that the cold-induced increase in polyunsaturated TAGs was largely prevented by blocking adipose triglyceride lipase.

Interpretation: We interpret these findings as that cold exposure feeds thermogenic tissues with TAG-derived fatty acids for combustion, resulting in a decrease of circulating TAG species, followed by increased hepatic production of polyunsaturated TAG species induced by liberation of free fatty acids stemming from adipose tissue.

Funding: This work was supported by the Netherlands CardioVascular Research Initiative: 'the Dutch Heart Foundation, Dutch Federation of University Medical Centers, the Netherlands Organisation for Health Research and Development and the Royal Netherlands Academy of Sciences' [CVON2017-20 GENIUS-II] to Patrick C.N. Rensen. Borja Martinez-Tellez is supported by individual postdoctoral grant from the Fundación Alfonso Martin Escudero and by a Maria Zambrano fellowship by the Ministerio de Universidades y la Unión Europea - NextGenerationEU (RR_C_2021_04). Lucas Jurado-Fasoli was supported by an individual pre-doctoral grant from the Spanish Ministry of Education (FPU19/01609) and with an Albert Renold Travel Fellowship from the European Foundation for the Study of Diabetes (EFSD). Martin Giera was partially supported by NWO XOmics project #184.034.019.

Keywords: Cold; Desaturation; Fatty acid metabolism; Intracellular lipolysis; Long-chain polyunsaturated fatty acids.

Conflict of interest statement

Declaration of interests None.

Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.

Figures

Fig. 1
Fig. 1
Study design and lipidomic changes during cold exposure. (a) Overview of the crossover study design. During the first occasion, participants were exposed to a personalized cooling protocol. During the other occasion, participants were exposed to constant room temperature. Under both conditions, venous blood was drawn at five sequential timepoints (i.e., baseline, 30, 60, 90, and 120 min) to obtain serum for longitudinal lipidomic profiling. (b) Dynamic changes in total free fatty acid and triacylglycerol concentration during cold exposure, as measured with enzymatic assays. Data is presented as mean and standard error of the mean. P values are obtained from ANOVA repeated measures. ∗P < 0.05, ∗∗P < 0.01; compared to baseline (i.e., 0 min). (c) Baseline relative abundance of the number of lipid classes. (d) Dynamic changes in lipid classes during cold exposure. Data is presented as the log2 fold change (log2FC) relative to baseline (i.e., 0 min). P values are obtained from ANOVA repeated measures. (e) Volcano plot showing the change of individual lipid species after 120 min of cold exposure. The X-axis shows the log2FC between 120 min of cold exposure vs. baseline, the Y-axis shows the P value. P values are obtained from paired Student's t test. Values between brackets indicate the absolute number and the percentage of lipid species within the lipid class that were modulated by cold exposure. (f) Heatmap of free fatty acid (FFA) species that significantly dynamically changed during cold exposure. The color of the squares represents the log2FC relative to baseline. FFAs are sorted by the number of double bonds. ANOVA repeated measures were used to compare the different timepoints. CE, cholesteryl esters; CER, ceramides; DAG, diacylglycerols; DCER, dihydroceramides; FFA, free fatty acids; FC, fold change; HCER, hexosylceramides; LCER, lactosylceramides; LPC, lysophosphatidylcholine; LPE, lysophosphatidylethanolamine; PC, phosphatidylcholine; PE, phosphatidylethanolamine; SM, sphingomyelin; TAG, triacylglycerols; TN, thermoneutral.
Fig. 2
Fig. 2
Triacylglycerol (TAG) species response to cold exposure. (a) Heatmap of TAG species that significantly dynamically changed during cold exposure. The color of the squares represents the log2 fold change (log2FC) relative to baseline. TAGs are sorted by the number of double bonds. ANOVA repeated measures were used to compare the different timepoints. (b) Bubble plot showing the P value (in color) and log2FC (on y-axis) after 120 min of cold exposure per TAG species categorized by the number of double bonds. P values are obtained from paired Student's t test. (c) Bubble plot showing the P value (in color) and log2FC (on y-axis) after 120 min of cold exposure per TAG species categorized by the total length of the TAG. P values are obtained from paired Student's t test. FA, fatty acid; FC, fold change; TAG, triacylglycerol.
Fig. 3
Fig. 3
Triacylglycerol (TAG) species' response to mild cold exposure in mice either treated with vehicle or with Atglistatin. (a and b) Bubble plots showing the P value (in color) and log2 fold change (log2FC; on y-axis) between mice exposed to cold and mice kept at thermoneutrality (n = 6 each group; both treated with vehicle) per TAG species categorized by the number of double bonds (a) and total length (b). P values are obtained from paired Student's t test. (c and d) Bubble plots showing the P value (in color) and log2FC (on y-axis) between mice exposed to cold and treated with Atglistatin and mice kept at thermoneutrality (n = 6 each group; treated with vehicle) per TAG species categorized by the number of double bonds (c) and total length (d). P values are obtained from paired Student's t test. FC, fold change; TAG, triacylglycerol.
Fig. 4
Fig. 4
Graphic summary of proposed mechanism. We propose a mechanism by which cold exposure induces intracellular lipolysis in white adipose tissue, increasing circulating free fatty acids that can subsequently drive hepatic biosynthesis and release of triacylglycerol (TAG)-containing very-low density lipoprotein (VLDL) particles to feed thermogenic tissues. The cold-activated thermogenic tissues (i.e., brown adipose tissue and skeletal muscle) take up TAG-derived fatty acids from VLDL particles as fuel for oxidation to generate heat. FA, fatty acid; FFA, free fatty acids; PUFA, polyunsaturated fatty acid; TAG, triacylglycerol; VLDL, very-low density lipoprotein.

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