Dataset Information

MGAT3 and MGAT5 overexpression alters the protein cargo of extracellular vesicles released by metastatic melanoma cells

ABSTRACT: Extracellular vesicles (EVs) are potential non-invasive diagnostic, prognostic, and therapeutic tools. Additionally, they are important contributors to tumorigenesis. Glycosylation has been found to modulate the composition of the EV proteome, and increased amounts of beta 1,6-branched N-glycans, synthesized by N-acetylglucosaminyltransferase V (GnT-V), are most commonly observed in melanoma and associated with decreased cell adhesion and increased metastasis. The opposite effect is caused by the addition of bisecting GlcNAc by N-acetylglucosaminyltransferase III (GnT-III). To date, the impact of these enzymes on EV cargo in melanoma remains unexplored. Methods: Flow cytometry was used to study the surface glycosylation of genetic variants of WM266-4 melanoma cells with induced overexpression of GnT-III or GnT-V encoding genes (MGAT3 or MGAT5) and EVs released by these cells. LC-MS/MS proteomics was applied to analyze the effect of altered glycosylation on the proteome of released EVs, followed by thorough bioinformatic analysis. Results: Flow cytometry analysis showed dynamic changes in surface glycosylation of EVs derived from melanoma cells overexpressing MGAT3 or MGAT5. Induced MGAT3 or MGAT5 overexpression also resulted in significant changes in EVs proteome. Proteomic analysis identified the total number of 1770 microvesicular and 704 exosomal proteins that play diverse roles in melanoma progression, including those with established diagnostic/prognostic potential and those closely linked to melanoma onset. Conclusions: Proteomic profiling of EVs derived from MGAT3 and MGAT5 overexpressing cells revealed functional shifts in EV protein content driven by glycosylation modifications. The study presented a potential multidimensional use of melanoma-derived EVs for diagnostic and prognostic purposes.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Malgorzata Przybylo