title: Proteomics-based metabolic modeling reveals that fatty acid oxidation (FAO) controls endothelial cell (EC) permeability.
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.
authors.affiliation: From the ‡Vascular Proteomics Lab.
authors.forename: Francesca
authors.initials: F
authors.lastname: Patella
authors.affiliation: §Apoptosis and Tumour Metabolism Lab.
authors.forename: Zachary T
authors.initials: ZT
authors.lastname: Schug
authors.affiliation: ¶The Blavatnik School of Computer Science, Tel Aviv University, 69978 Tel Aviv, Israel; ‖School of Physics and Astronomy, Tel Aviv University, 69978 Tel Aviv, Israel;
authors.forename: Erez
authors.initials: E
authors.lastname: Persi
authors.affiliation: From the ‡Vascular Proteomics Lab.
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Neilson
authors.affiliation: **Tumour Cell Migration Lab, Cancer Research UK Beatson Institute, Switchback Road, G61 1BD, Glasgow, UK;
authors.forename: Zahra
authors.initials: Z
authors.lastname: Erami
authors.affiliation: ‡‡Department of Life Sciences and Systems Biology, University of Torino, Via Accademia Albertina 13, 10123 Torino, Italy;
authors.forename: Daniele
authors.initials: D
authors.lastname: Avanzato
authors.affiliation: §§Laboratory of Transgenic Mouse Models, Candiolo Cancer Institute - FPO, IRCCS, Str prov 142 Km 3.95, 10060, Candiolo, Torino, Italy; ¶¶Department of Science and Drug Technology, University of Torino, Via P. Giuria, 9 - 10125 Torino, Italy;
authors.forename: Federica
authors.initials: F
authors.lastname: Maione
authors.affiliation: From the ‡Vascular Proteomics Lab.
authors.forename: Juan R
authors.initials: JR
authors.lastname: Hernandez-Fernaud
authors.affiliation: §Apoptosis and Tumour Metabolism Lab.
authors.forename: Gillian
authors.initials: G
authors.lastname: Mackay
authors.affiliation: §Apoptosis and Tumour Metabolism Lab.
authors.forename: Liang
authors.initials: L
authors.lastname: Zheng
authors.affiliation: From the ‡Vascular Proteomics Lab.
authors.forename: Steven
authors.initials: S
authors.lastname: Reid
authors.affiliation: ‖‖MRC Cancer Unit, Cambridge Biomedical Campus, University of Cambridge, Hutchison/MRC Research Centre, Box 197, CB2 0XZ, Cambridge, UK;
authors.forename: Christian
authors.initials: C
authors.lastname: Frezza
authors.affiliation: §§Laboratory of Transgenic Mouse Models, Candiolo Cancer Institute - FPO, IRCCS, Str prov 142 Km 3.95, 10060, Candiolo, Torino, Italy; ¶¶Department of Science and Drug Technology, University of Torino, Via P. Giuria, 9 - 10125 Torino, Italy;
authors.forename: Enrico
authors.initials: E
authors.lastname: Giraudo
authors.affiliation: ‡‡Department of Life Sciences and Systems Biology, University of Torino, Via Accademia Albertina 13, 10123 Torino, Italy;
authors.forename: Alessandra
authors.initials: A
authors.lastname: Fiorio Pla
authors.affiliation: **Tumour Cell Migration Lab, Cancer Research UK Beatson Institute, Switchback Road, G61 1BD, Glasgow, UK;
authors.forename: Kurt
authors.initials: K
authors.lastname: Anderson
authors.affiliation: ¶The Blavatnik School of Computer Science, Tel Aviv University, 69978 Tel Aviv, Israel; Sackler School of Medicine, and Department of Molecular Microbiology and Biotechnology, Faculty of Life Sciences, Tel-Aviv University, 69978 Tel Aviv, Israel.
authors.forename: Eytan
authors.initials: E
authors.lastname: Ruppin
authors.affiliation: §Apoptosis and Tumour Metabolism Lab.
authors.forename: Eyal
authors.initials: E
authors.lastname: Gottlieb
authors.affiliation: From the ‡Vascular Proteomics Lab, s.zanivan@beatson.gla.ac.uk.
authors.forename: Sara
authors.initials: S
authors.lastname: Zanivan
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002334:Carnitine O-Palmitoyltransferase / Q000037:antagonists & inhibitors*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004852:Epoxy Compounds / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D066298:In Vitro Techniques; D055442:Metabolome; D051379:Mice; D008954:Models, Biological; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010539:Permeability; D040901:Proteomics / Q000379:methods*
chemical_list: D004852:Epoxy Compounds; D005227:Fatty Acids; D000255:Adenosine Triphosphate; C577474:CPT1A protein, human; D002334:Carnitine O-Palmitoyltransferase; C054207:etomoxir