Using the Zebrafish Embryo to Dissect the Early Steps of the Metastasis Cascade

Abstract

Most cancers end up with the death of patients caused by the formation of secondary tumors, called metastases. However, how these secondary tumors appear and develop is only poorly understood. A fine understanding of the multiple steps of the metastasis cascade requires in vivo models allowing high spatiotemporal analysis of the behavior of metastatic cells. Zebrafish embryos combine several advantages such as transparency, small size, stereotyped anatomy, and easy handling, making it a very powerful model for cell and cancer biology, and in vivo imaging analysis. In the following chapter, we describe a complete procedure allowing in vivo imaging methods, at high throughput and spatiotemporal resolution, to assess the behavior of circulating tumor cells (CTCs) in an experimental metastasis assay. This protocol provides access, for the first time, to the earliest steps of tumor cell seeding during metastasis formation.

Zika virus enhances monocyte adhesion and transmigration favoring viral dissemination to neural cells

“…Since transmigration is a very challenging process to study in mammals, we tested whether zebrafish embryos could represent a suitable alternative. To this end, we developed a novel quantitative xenotypic system based on a procedure designed to dissect the early behavior of circulating cells in the zebrafish embryo bloodstream 40,41 . Monocytes were infected with ZIKV alone or ZIKV in the presence of a pan-flavivirus antibody known to increase infection through antibody-dependent enhancement (ADE) 42,43 (Supplementary Fig.…”

Towards Resolving the Pro- and Anti-Tumor Effects of the Aryl Hydrocarbon Receptor

“…Studies in the zebrafish model are consistent with a role for the AHR in metastasis in vivo. While the zebrafish model has significant advantages over other models of cell migration (e.g., zebrafish contain a full complement of extracellular matrix proteins that interact with and influence migratory cells and are easy to image, and the model is less expensive and more rapid than other animal models [ 78 , 82 , 121 , 122 ]) some caution should be exercised when extrapolating zebrafish results to human systems. The evolutionary distance between zebrafish and humans creates some uncertainty in interpretation of the results.…”

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

“…Pre-labelled EVs, polystyrene beads (Phosphorex) or tumors cells were injected with a Nanoject microinjector 2 (Drummond) and microforged glass capillaries (25 to 30 mm inner diameter) filled with mineral oil (Sigma). 27,6 nL of a EV, beads or cell suspension (at 100.10 6 cells) per ml were injected into the duct of Cuvier of the embryos under the M205 FA stereomicroscope (Leica), as previously described (Follain et al, 2018b;Stoletov et al, 2010). For the priming experiments, 32hpf embryos were injected with either Zmel1 EVs or 100nm polystyrene beads (together with fluorescent dextran to assess the efficiency of injection).…”