Journal Article

DZNE-2025-00441

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Small molecule inhibitors of hnRNPA2B1-RNA interactions reveal a predictable sorting of RNA subsets into extracellular vesicles.

Corsi, J. ; Semnani, P. S. ; Peroni, D. ; Belli, R. ; Morelli, A. ; Lassandro, M. ; Sidarovich, V. ; Adami, V. ; Valentini, C. ; Cavallerio, P. ; Grosskreutz, J. ; Fabbiano, F. ; Grossmann, D. ; Hermann, A.DZNE* ; Tell, G. ; Basso, M. ; D'Agostino, V. G.

2025
Oxford Univ. Press Oxford

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Please use a persistent id in citations: doi:10.1093/nar/gkaf176

Abstract: 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.

Keyword(s): Humans (MeSH) ; Extracellular Vesicles: metabolism (MeSH) ; Heterogeneous-Nuclear Ribonucleoprotein Group A-B: metabolism (MeSH) ; Heterogeneous-Nuclear Ribonucleoprotein Group A-B: genetics (MeSH) ; RNA: metabolism (MeSH) ; RNA: genetics (MeSH) ; Protein Binding (MeSH) ; Proteomics: methods (MeSH) ; HEK293 Cells (MeSH) ; NF-kappa B: metabolism (MeSH) ; Small Molecule Libraries: pharmacology (MeSH) ; Heterogeneous-Nuclear Ribonucleoprotein Group A-B ; RNA ; hnRNP A2 ; NF-kappa B ; Small Molecule Libraries

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Contributing Institute(s):

1. Translational Neurodegeneration (AG Hermann)

Research Program(s):

1. 353 - Clinical and Health Care Research (POF4-353) (POF4-353)

Appears in the scientific report 2025

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