Abstract B17: In-depth proteomics unveils fatty acid oxidation role in controlling vascular permeability

Abstract

Abstract Endothelial cells (ECs) play a key role in maintaining vascular functionality. Alterations in vessel stability and permeability are hallmarks of tumor angiogenesis and may affect the efficacy of anti-cancer therapy. Modulating EC metabolism has emerged as a promising strategy to target angiogenesis in pathological conditions, but so far, little is known about the role of EC metabolism in the regulation of vessel stability and permeability. In this work we took advantage of a simplified in-vitro model of angiogenesis, where HUVECs (human umbilical vein endothelial cells) form a vascular-like network on a tridimensional matrix, to elucidate the role of EC metabolism during this process. By integrating high resolution proteomic data with a human genome scale metabolic model we built up the first predictive model of metabolic fluxes occurring in ECs forming a complex network and identified increased fluxes for reactions involving fatty acid oxidation (FAO) enzymes when ECs are assembled into a fully formed network. Upon inhibition of CPT1, the FAO rate-limiting enzyme, we disrupted the network and reduced cellular ATP levels and oxygen consumption, which were restored by replenishing the tricarboxylic acid cycle (TCAc). Remarkably, phosphoproteomic changes measured upon CPT1A inhibition evoked those triggered by thrombin, a potent inducer of EC permeability through calcium signaling. Indeed, acute CPT1A inhibition increased EC permeability in-vitro and leakage of fully formed blood vessel in-vivo, which were restored by replenishing the TCAc or inhibiting calcium influx. FAO emerges as central regulator of blood vessel stability, revealing the possibility of targeting FAO to interfere with tumor vessel permeability. Citation Format: Francesca Patella, Zachary T. Schug, Erez Persi, Lisa J. Neilson, Zahra Erami, Daniele Avanzato, Federica Maione, Juan R. Hernandez-Fernaud, Gillian Mackay, Liang Zheng, Steven Reid, Christian Frezza, Enrico Giraudo, Alessandra Fiorio Pla, Kurt Anderson, Eytan Ruppin, Eyal Gottlieb, Sara Zanivan. In-depth proteomics unveils fatty acid oxidation role in controlling vascular permeability. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Angiogenesis and Vascular Normalization: Bench to Bedside to Biomarkers; Mar 5-8, 2015; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl):Abstract nr B17.