Ricerc@Sapienza

Crosstalk between cancer cells and the niche deeply influences self-renewal, plasticity and metastasis and Extracellular Vesicles (EVs) are considered pivotal in cellular intercommunication also in these contexts.
This project builds on our findings showing that the RNA-Binding-Protein (RBP) SYNCRIP impacts on exosome miRNA content. Furthermore, we identified a miRNA extra-seed motif sufficient to determine SYNCRIP-mediated exosome loading. These results pave the way for novel therapeutic approaches based on the in vivo production of exosomes containing ncRNAs engineered to be loaded into exosomes, to modulate gene expression in a customized way. Moreover, our preliminary results shed light on the role of other RBPs in the regulation of miRNA exosome loading. Thus, we here propose the identification of others RBPs and consensus motifs in the hypothesis that they may control specific miRNA sorting in the EMT, a transdifferentiation process required for the metastasis of epithelial cancers.
The proposed aims are 1-to validate the RBPs impact on cells and tumor niche during the Epithelial-to-Mesenchymal Transition (EMT), by controlling miRNA EVs loading; 2-to reconduct the EVs pro- or anti-EMT activity to specific subsets of miRNAs and to engineer ncRNAs with specific motifs to vehiculate them into EVs; 3-to unveil the epitranscriptomic modifications of differentially exported miRNAs and to explore their functional role.
We expect to dissect the functional role of EVs in cell communication in EMT, in relation to presence/absence of specific RBPs; to identify specific subsets of differentially exported/cell retained miRNAs and conserved sequence motifs as RBPs binding sites; to clarify the role of m6A RNA modification in controlling RBPs-mediated miRNA sorting in EVs.
The progressive understanding of the loading machinery and of the mechanisms involved in ncRNAs compartmentalization within EVs, hold strong promise for further selective miRNA EVs cargo loading.