High-resolution targeted mass spectrometry for comprehensive quantification of sphingolipids: clinical applications and characterization of extracellular vesicles

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Felippe, Thiago V. D. ; Toro, Diana M. ; de Carvalho, Jonatan C. S. ; Nobre-Azevedo, Pedro ; Rodrigues, Luiz F. M. ; Oliveira, Bianca T. M. ; da Silva-Neto, Pedro V. ; Vilela, Adriana F. L. ; Almeida, Fausto ; Faccioli, Lucia H. ; Sorgi, Carlos A.

• Document type: Journal article
• Source: Analytical Biochemistry; v. 698, p. 12-pg., 2024-12-04.

Abstract

Sphingolipids (SL), a class of membrane lipids, play important roles in numerous biological processes. Their significant structural diversity poses challenges for accurate quantification. To address this, liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has emerged as a powerful tool for sphingolipidomics, capable of profiling these lipids comprehensively. In this study, we utilized LC-MS/MS with high-resolution mass spectrometry (MRMHR) to develop a targeted method for the identification and quantification of various SL species. This method, based on validated parameters such as precursor/fragment ions (m/z) and retention time, demonstrated high sensitivity and accuracy, successfully identifying SL species across 12 distinct classes. Its open-panel design also facilitates the analysis of new SL-species targets. Notably, using this approach, we identified 40 SL species in plasma samples from COVID-19 patients, and we determined the influence of matrix metalloproteinase-3 (MMP-3) expression on the positive downstream of SL metabolism. Beyond plasma analysis, this method has potential applications in other biomedical contexts, such as extracellular vesicles (EVs), describing the cargo of sphingosine-1-phosphate (S1P) on macrophage-derived EVs. The establishment of this targeted workflow enabling precise quantification of a wide range of SL species, holds promise for identifying novel biomarkers and therapeutic targets. (AU)

• FAPESP's process: 24/03414-5 - Integrated Analysis of Sphingolipid Metabolism and Extracellular Particle Composition in the Context of a Hight-Fat-Diet: Implications for Lipoproteins, Vesicle Biogenesis, and the Role in Macrophage Response to COVID-19
• Grantee: Carlos Arterio Sorgi
• Support Opportunities: Regular Research Grants
• FAPESP's process: 21/04590-3 - Exploratory analysis of functional lipidomics in the COVID-19 pathophysiology: molecular targets, metabolism and biochemical reprogramming due to hypoxia
• Grantee: Carlos Arterio Sorgi
• Support Opportunities: Regular Research Grants
• FAPESP's process: 23/07776-6 - Evaluation of the therapeutic role of selective serotonin reuptake inhibitor antidepressants in modulating the inflammatory response and sphingolipid metabolism in SARS-CoV-2 infected macrophages
• Grantee: Jonatan Constança Silva de Carvalho
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 14/07125-6 - New functional aspects of eicosanoids
• Grantee: Lúcia Helena Faccioli
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 20/05207-6 - Prospective evaluation of lipid mediators production on the immune response against COVID-19: search for biomarkers and new therapeutic targets in the evolution of the disease
• Grantee: Lúcia Helena Faccioli
• Support Opportunities: Regular Research Grants
• FAPESP's process: 15/00658-1 - New functional aspects of eicosanoids
• Grantee: Lúcia Helena Faccioli
• Support Opportunities: Multi-user Equipment Program