GlycoRNA complexed with heparan sulfate regulates VEGF-A signalling

Receptor-ligand interactions govern a wide array of biological pathways, facilitating a cell's ability to interrogate and integrate information from the extracellular space. Here, we identify heparan sulfate proteoglycans (HSPGs) as a major organizational mechanism of glycoRNA and cell surface RNA binding proteins (csRBPs). If growth factors, which use HSPGs to bind the cell surface, interact with RNA is not well understood. Here we use biochemical pulldowns of VEGF-A165 coupled to RNA-seq to determine if and which RNAs are selectively isolated with this growth factor in vitro. Overall design: Small RNA fractions (generally less than 200 nucleotides) were biochemically isolated from HUVEC cells. This RNA served as input material for RNA-seq libraries as well as to perform a biochemical enrichment with anti-VEGF-A antibodies with or without the addition of VEGF-A165. After incubation with the small RNA from HUVEC cells, enriched RNAs bound to the antibody or antibody-VEGF-A165 complex were purified and RNA-seq libraries generated from the resulting RNA.