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Studying the Fate of Tumor Extracellular Vesicles at High Spatiotemporal Resolution Using the Zebrafish Embryo.

Developmental cell (2019-02-13)
Vincent Hyenne, Shima Ghoroghi, Mayeul Collot, Joanna Bons, Gautier Follain, Sébastien Harlepp, Benjamin Mary, Jack Bauer, Luc Mercier, Ignacio Busnelli, Olivier Lefebvre, Nina Fekonja, Maria J Garcia-Leon, Pedro Machado, François Delalande, Ana Amor López, Susana Garcia Silva, Frederik J Verweij, Guillaume van Niel, Farida Djouad, Héctor Peinado, Christine Carapito, Andrey S Klymchenko, Jacky G Goetz
ABSTRACT

Tumor extracellular vesicles (EVs) mediate the communication between tumor and stromal cells mostly to the benefit of tumor progression. Notably, tumor EVs travel in the bloodstream, reach distant organs, and locally modify the microenvironment. However, visualizing these events in vivo still faces major hurdles. Here, we describe an approach for tracking circulating tumor EVs in a living organism: we combine chemical and genetically encoded probes with the zebrafish embryo as an animal model. We provide a first description of tumor EVs' hemodynamic behavior and document their intravascular arrest. We show that circulating tumor EVs are rapidly taken up by endothelial cells and blood patrolling macrophages and subsequently stored in degradative compartments. Finally, we demonstrate that tumor EVs activate macrophages and promote metastatic outgrowth. Overall, our study proves the usefulness and prospects of zebrafish embryo to track tumor EVs and dissect their role in metastatic niches formation in vivo.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Fibronectin bovine plasma, solution, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
PKH26 Red Fluorescent Cell Linker Mini Kit for General Cell Membrane Labeling, Distributed for Phanos Technologies