Molecular characterization of exosomes in the Medullary Thyroid Carcinoma cell line TT

Abstract

Medullary Thyroid Carcinoma (MTC) is a rare type among tumors affecting the gland, responding for about 5% of cases, yet causing nearly 13% of total deaths. As of now, there is no information in the literature of whether or not MTCs secrete exosomes, or miRNA, although many malignancies have been characterized in this aspect, uncovering an important role for exosome secretion in mediating mechanisms of tumorigenesis. Our goal is to investigate whether MTC-lineage cells secrete exosomes and to characterize exosomal content. Exosomes will be purified by ordinary centrifugation in a commercially available polymer designed to allow separation by density and size (Exo-spin), a broadly accepted method. Once exosomes are purified, their morphological characterization will be obtained by electron microscopy and nanoparticle tracking analysis. Purified and validated exosomes will be further characterized regarding their miRNA content, with Nanostring nCounter technology, and these data will be used to predict target pathways of secreted miRNAs, using the online tools miRWalk and PANTHER. With the aid of these results, we expect to be able to predict interactions among MTC and other cells in the tumor microenvironment, paving the way for future experimental studies. (AU)