%0 Journal Article
%A Corsi, Jessica
%A Semnani, Pouriya Sharbatian
%A Peroni, Daniele
%A Belli, Romina
%A Morelli, Alessia
%A Lassandro, Michelangelo
%A Sidarovich, Viktoryia
%A Adami, Valentina
%A Valentini, Chiara
%A Cavallerio, Paolo
%A Grosskreutz, Julian
%A Fabbiano, Fabrizio
%A Grossmann, Dajana
%A Hermann, Andreas
%A Tell, Gianluca
%A Basso, Manuela
%A D'Agostino, Vito G
%T Small molecule inhibitors of hnRNPA2B1-RNA interactions reveal a predictable sorting of RNA subsets into extracellular vesicles.
%J Nucleic acids research
%V 53
%N 5
%@ 0305-1048
%C Oxford
%I Oxford Univ. Press
%M DZNE-2025-00441
%P gkaf176
%D 2025
%X Extracellular vesicles (EVs) are cell-secreted membranous particles contributing to intercellular communication. Coding and noncoding RNAs can be detected as EV cargo, and RNA-binding proteins (RBPs), such as hnRNPA2B1, have been circumstantially implicated in EV-RNA sorting mechanisms. However, the contribution of competitive RBP-RNA interactions responsible for RNA-sorting outcomes is still unclear, especially for predicting the EV-RNA content. We designed a reverse proteomic analysis exploiting the EV-RNA to identify intracellular protein binders in vitro. Using cells expressing a recombinant hnRNPA2B1 to normalize competitive interactions, we prioritized a network of heterogeneous nuclear ribonucleoproteins and purine-rich RNA sequences subsequently validated in secreted EV-RNA through short fluorescent RNA oligos. Then, we designed a GGGAG-enriched RNA probe that efficiently interacted with a full-length human hnRNPA2B1 protein. We exploited the interaction to conduct a pharmacological screening and identify inhibitors of the protein-RNA binding. Small molecules were orthogonally validated through biochemical and cell-based approaches. Selected drugs remarkably impacted secreted EV-RNAs and reduced an RNA-dependent, EV-mediated paracrine activation of NF-kB in recipient cells. These results demonstrate the relevance of post-transcriptional mechanisms for EV-RNA sorting and the possibility of predicting the EV-RNA quality for developing innovative strategies targeting discrete paracrine functions.
%K Humans
%K Extracellular Vesicles: metabolism
%K Heterogeneous-Nuclear Ribonucleoprotein Group A-B: metabolism
%K Heterogeneous-Nuclear Ribonucleoprotein Group A-B: genetics
%K RNA: metabolism
%K RNA: genetics
%K Protein Binding
%K Proteomics: methods
%K HEK293 Cells
%K NF-kappa B: metabolism
%K Small Molecule Libraries: pharmacology
%K Heterogeneous-Nuclear Ribonucleoprotein Group A-B (NLM Chemicals)
%K RNA (NLM Chemicals)
%K hnRNP A2 (NLM Chemicals)
%K NF-kappa B (NLM Chemicals)
%K Small Molecule Libraries (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:40103230
%R 10.1093/nar/gkaf176
%U https://pub.dzne.de/record/277552