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 tridimensional matrix and organize into a vascular-like network. We discovered how fatty acid oxidation increases when ECs are assembled into a fully formed network that can be disrupted by inhibiting CPT1A, the fatty acid oxidation rate-limiting enzyme. Acute CPT1A inhibition reduces cellular ATP levels and oxygen consumption, which are restored by replenishing the tricarboxylic acid cycle. 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 tricarboxylic acid cycle. Fatty acid oxidation emerges as central regulator of endothelial functions and blood vessel stability and druggable pathway to control pathological vascular permeability.

Proteomics-based metabolic modelling reveals that fatty acid oxidation controls endothelial cell permeability.

AVANZATO, DANIELE;MAIONE, FEDERICA;GIRAUDO, Enrico;FIORIO PLA, ALESSANDRA;
2015

Abstract

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 tridimensional matrix and organize into a vascular-like network. We discovered how fatty acid oxidation increases when ECs are assembled into a fully formed network that can be disrupted by inhibiting CPT1A, the fatty acid oxidation rate-limiting enzyme. Acute CPT1A inhibition reduces cellular ATP levels and oxygen consumption, which are restored by replenishing the tricarboxylic acid cycle. 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 tricarboxylic acid cycle. Fatty acid oxidation emerges as central regulator of endothelial functions and blood vessel stability and druggable pathway to control pathological vascular permeability.
14
3
621
634
https://pubmed.ncbi.nlm.nih.gov/25573745/
Patella F; Schug ZT; Persi E; Neilson LJ; Erami Z; Avanzato D; Maione F; Hernandez-Fernaud JR; Mackay G; Zheng L; Reid S; Frezza C; Giraudo E; Fiorio Pla A; Anderson K; Ruppin E; Gottlieb E; Zanivan S.
File in questo prodotto:
File Dimensione Formato  
Mol Cell Proteomics-2015-Patella-621-34.pdf

Accesso riservato

Tipo di file: PDF EDITORIALE
Dimensione 881.54 kB
Formato Adobe PDF
881.54 kB Adobe PDF   Visualizza/Apri   Richiedi una copia
Patella et al._PostPrintlast.pdf

Accesso aperto

Descrizione: Post Print
Tipo di file: POSTPRINT (VERSIONE FINALE DELL’AUTORE)
Dimensione 1.76 MB
Formato Adobe PDF
1.76 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/151936
Citazioni
  • ???jsp.display-item.citation.pmc??? 31
  • Scopus 65
  • ???jsp.display-item.citation.isi??? 68
social impact