The cancer microenvironment plays a critical role in tumour growth and metastasis. Recent discoveries have pointed out that certain immune cells have a “pro-tumour” function. It is important to identify these functions and implement them into a more effective anti-cancer therapy. This project proposes to use the zebrafish cancer models to gain a better understanding of the role of innate immune cells during metastasis formation.

 

The specific goal of this project is to inhibit the process of metastasis by blocking markers on the innate immune cells. The zebrafish is an excellent model to this aim because it is perfectly applicable to visualize fluorescent immune and human cells during cancer development. Fluorescently marked human breast cancer cells in red will be injected into transgenic zebrafish embryos with green fluorescent neutrophils or macrophages. The exact role of neutrophils and macrophages in the metastatic process will be determined via chemical and genetic ablation of these immune cells. For identification of cancer-induced genes on the immune cells and the host microenvironment, the fluorescent immune cells will be isolated by FACS from embryos with and without tumours. The gene expression patterns of the immune and host cells will be determined using RNAseq and compared to already describe human genes expressed on tumor-associated-neutrophils (TANs) and tumor-associated-macrophages (TAMs). Novel candidate genes will be selected and their function validated using genetic knock out approach and tested in the zebrafish metastatic model. Promising genes will be validated in the mouse model and eventually considered for anti-cancer treatment.

• He S, Lamers GEM, Beenakker JWM, Cui C, Ghotra VPS, Danen EHJ, Meijer AH, Spaink HP and Snaar-Jagalska BE (2012) Neutrophil-mediated experimental metastasis is enhanced by VEGFR inhibition in a zebrafish xenograft model. J. Pathology 227: 431-45
• Tulotta C, Stefanescu C, Beletkaia E, Busmann J, Schmidt T and  Snaar-Jagalska BE (2016) Inhibition of signaling between human CXCR4 and zebrafish ligands by the small molecule IT1t impairs the formation of triple-negative breast cancer early metastases in a zebrafish xenograft. Dis. Model Mech. 9: 141–153
• Tulotta C, He S, van der Ent W, Chen L, Groenewoud A, Spaink HP, Snaar-Jagalska BE (2016) Imaging Cancer Angiogenesis and Metastasis in a Zebrafish Embryo Model. Adv Exp Med Biol. 2016;916:239-63
• Tulotta C, He S, Chen L, Groenewoud A, van der Ent W, Meijer AH, Spaink HP, Snaar-Jagalska BE (2016) Imaging of Human Cancer Cell Proliferation, Invasion, and Micrometastasis in a Zebrafish Xenogeneic Engraftment Model. Methods Mol Biol. 2016;1451:155-69

• bioactive molecules
• cancer microenvironment
• development & disease
• drug discovery
• zebrafish xenografts