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Proteomic analysis of signaling pathways modulated by FABP5 in macrophages.
Research Square 2023 September 13
Background While acute inflammation serves essential functions in maintaining tissue homeostasis, chronic inflammation is causally linked to many diseases. Macrophages are a major cell-type that orchestrates inflammatory processes. During inflammation, macrophages undergo polarization and activation, thereby mobilizing pro-inflammatory and anti-inflammatory transcriptional programs that regulate ensuing macrophage functions. Fatty acid binding protein 5 (FABP5) is a lipid chaperone that is highly expressed in macrophages. FABP5 deletion is implicated in driving macrophages towards an anti-inflammatory phenotype, yet the signaling pathways regulated by macrophage FABP5 have not been systematically profiled. Herein, we leveraged proteomic and phosphoproteomic approaches to characterize pathways modulated by FABP5 in M1 and M2 polarized bone marrow derived macrophages (BMDMs). Results Stable isotope labeling by amino acids (SILAC) based analysis of M1 and M2 polarized wild-type (WT) and FABP5 knockout (KO) BMDMs revealed numerous differentially regulated proteins and phosphoproteins. FABP5 deletion impacted several downstream pathways associated with inflammation, cytokine production, oxidative stress, and kinase activity. Kinase enrichment analysis based on phosphorylated sites revealed key kinases, including members of the GRK family, that were altered in FABP5 KO BMDMs. Reactive oxygen species (ROS) levels were elevated in M1 polarized KO macrophages, consistent with the differential protein expression profiles. Conclusions This study represents a comprehensive characterization of the impact of FABP5 deletion upon the proteomic and phosphoproteomic landscape of M1 and M2 polarized BMDMs. Loss of FABP5 altered multiple pathways implicated in inflammatory responses and macrophage function. This work provides a foundation for future studies seeking to investigate the therapeutic potential of FABP5 inhibition in pathophysiological states resulting from dysregulated inflammatory signaling.
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