Multi-Omic Profiling of Macrophages Treated with Phospholipids Containing Omega-3 and Omega-6 Fatty Acids Reveals Complex Immunomodulatory Adaptations at Protein, Lipid and Metabolic Levels

Abstract

In recent years, several studies have demonstrated that polyunsaturated fatty acids have strong immunomodulatory properties, altering several functions of macrophages. In the present work, we sought to provide a multi-omic approach combining the analysis of the lipidome, the proteome, and the metabolome of RAW 264.7 macrophages supplemented with phospholipids containing omega-3 (PC 18:0/22:6; ω3-PC) or omega-6 (PC 18:0/20:4; ω6-PC) fatty acids, alone and in the presence of lipopolysaccharide (LPS). Supplementation of macrophages with ω3 and ω6 phospholipids plus LPS produced a significant reprogramming of the proteome of macrophages and amplified the immune response; it also promoted the expression of anti-inflammatory proteins (e.g., pleckstrin). Supplementation with the ω3-PC and ω6-PC induced significant changes in the lipidome, with a marked increase in lipid species linked to the inflammatory response, attributed to several pro-inflammatory signalling pathways (e.g., LPCs) but also to the pro-resolving effect of inflammation (e.g., PIs). Finally, the metabolomic analysis demonstrated that supplementation with ω3-PC and ω6-PC induced the expression of several metabolites with a pronounced inflammatory and anti-inflammatory effect (e.g., succinate). Overall, our data show that supplementation of macrophages with ω3-PC and ω6-PC effectively modulates the lipidome, proteome, and metabolome of these immune cells, affecting several metabolic pathways involved in the immune response that are triggered by inflammation.

Metabolic and age-associated epigenetic barriers during direct reprogramming of mouse fibroblasts into induced cardiomyocytes

“…In both clusters, MEFs mock and MGT present significant differences in the abundance of metabolites, where, in some cases, MEFs MGT resemble HL-1 cells, as for increased phenylethylamine (Figure S1d), and decreased L-glutamine (Figure S1e) and adenine (Figure S1f). Differential analysis of cellular metabolites between mock and MGT-transduced MEFs was associated with 18 metabolic pathways constituted by the most differently expressed metabolites ( p -value < 0.05 ) (Figure 2c, S1g and Table S3) (Maurício et al, 2022). From this analysis, alanine, aspartate and glutamate metabolism ( p -value < 0.0001); phenylalanine metabolism ( p -value < 0.001); nicotinate and nicotinamide metabolism ( p -value < 0.01); phenylalanine, tyrosine and tryptophan biosynthesis ( p -value < 0.01) and taurine and hypotaurine metabolism ( p -value < 0.05) represent the most significant altered pathways (Figure 2c and S1g).…”

“…The mass spectrometer (MS) was operated as previously described. (Maurício et al, 2022) Positive and negative ion modes were employed (voltages 3.1 kV and -2.8 kV, respectively). The capillary temperature was set to 350 °C, and a sheath gas flow of 20 U was used.…”

Inhibiting SETD7 methyl‐transferase activity impairs differentiation, lipid metabolism and lactogenesis in mammary epithelial cells

“…[45], while peak assignment in data from C18 LC–MS/MS was performed using the bioinformatic tool msdial 4.70 [47], as described by Maurício et al . [48].…”

Local problems need global solutions: The metabolic needs of regenerating organisms

“…Although there is a significant interest in sugar and amino acid traffic, more recent work has re-started to look into lipids and uncovering novel roles in metabolism and signalling. [11][12][13] First, the nutritional flow is regulated by intestinal cells after food ingestion. To obtain energy from the dietary components, complex dietary carbohydrates, protein and lipids, are broken down to ease their intestinal absorption, cellular transport and secretion into circulation.…”

“…Upon food ingestion, the main role of the digestive system is to obtain lipids, carbohydrates, proteins, minerals and vitamins. Although there is a significant interest in sugar and amino acid traffic, more recent work has re‐started to look into lipids and uncovering novel roles in metabolism and signalling 11–13 . First, the nutritional flow is regulated by intestinal cells after food ingestion.…”