Proteomics-based metabolic modelling reveals that fatty acid oxidation controls endothelial cell permeability
نویسندگان
چکیده
FAO maintains endothelial permeability 3 Abbreviations: EC endothelial cell FAO fatty acid oxidation TCAc tricarboxylic acid cycle FA fatty acid iMAT integrative metabolic analysis tool GSMM genome-scale metabolic network model SILAC stable-isotope labeling with amino acids in cell culture HUVEC human umbilical vein endothelial cells ECM extracellular matrix TEER trans-endothelial electrical resistance DCA dichloroacetate VEGF vascular endothelial growth factor FAO maintains endothelial permeability 4 Summary Endothelial cells (ECs) play a key role to maintain the functionality of blood vessels. Altered EC permeability causes severe impairment in vessel stability and is a hallmark of pathologies such as cancer and thrombosis. Integrating label-free quantitative proteomics data into genome-wide metabolic modeling, we built up a model which predicts the metabolic fluxes in ECs when cultured on a tridimensional matrix and organize into a vascular-like network. We discovered how fatty acid oxidation (FAO) increases when ECs are assembled into a fully formed network that can be disrupted by inhibiting CPT1A, the FAO rate-limiting enzyme. Acute CPT1A inhibition reduces cellular ATP levels and oxygen consumption, which are restored by replenishing the tricarboxylic acid cycle (TCAc). Remarkably, global phosphoproteomic changes measured upon acute CPT1A inhibition pinpointed altered calcium signaling. Indeed, CPT1A inhibition increases intracellular calcium oscillations. Finally, inhibiting CPT1A induces hyperpermeability in-vitro and leakage of blood vessel in-vivo, which were restored blocking calcium influx or replenishing the TCAc. FAO emerges as central regulator of endothelial functions and blood vessel stability and druggable pathway to control pathological vascular permeability.
منابع مشابه
Proteomics-Based Metabolic Modeling Reveals That Fatty Acid Oxidation (FAO) Controls Endothelial Cell (EC) Permeability*□S
متن کامل
Proteomics-Based Metabolic Modeling Reveals That Fatty Acid Oxidation (FAO) Controls Endothelial Cell (EC) Permeability*
Endothelial cells (ECs) play a key role to maintain the functionality of blood vessels. Altered EC permeability causes severe impairment in vessel stability and is a hallmark of pathologies such as cancer and thrombosis. Integrating label-free quantitative proteomics data into genome-wide metabolic modeling, we built up a model that predicts the metabolic fluxes in ECs when cultured on a tridim...
متن کاملUntargeted metabolomics reveals distinct metabolic reprogramming in endothelial cells co-cultured with CSC and non-CSC prostate cancer cell subpopulations
Tumour angiogenesis is an important hallmark of cancer and the study of its metabolic adaptations, downstream to any cellular change, can reveal attractive targets for inhibiting cancer growth. In the tumour microenvironment, endothelial cells (ECs) interact with heterogeneous tumour cell types that drive angiogenesis and metastasis. In this study we aim to characterize the metabolic alteration...
متن کاملLinoleic acid and TNF-alpha cross-amplify oxidative injury and dysfunction of endothelial cells.
Factors implicated in the development of atherosclerosis include metabolic alterations of the endothelium induced by certain lipids and inflammatory cytokines. To study the hypothesis that the combined presence of unsaturated fatty acids and inflammatory cytokines may cross-amplify their individual injurious effects, cultured endothelial cells were treated with 90 mu M of linoleic acid (18:2 n-...
متن کاملFoxp3 drives oxidative phosphorylation and protection from lipotoxicity.
Tregs can adopt a catabolic metabolic program with increased capacity for fatty acid oxidation-fueled oxidative phosphorylation (OXPHOS). It is unclear why this form of metabolism is favored in Tregs and, more specifically, whether this program represents an adaptation to the environment and developmental cues or is "hardwired" by Foxp3. Here we show, using metabolic analysis and an unbiased ma...
متن کامل