Identification of mRNAS enriched in endothelial cell protrusions

Deciphering the complex biology of cell migration is key to understand human-related disorders. During angiogenesis, endothelial cells engage in coordinated migration events to form new blood vessels from parental counterparts. However, while subcellular localisation of mRNAs and localised translation are fundamental steps between gene transcription and protein activity, whether local regulation of gene expression controls the complex morphogenetic process of angiogenesis has never been addressed. Here, we set out to investigate the cytoplasmic distribution of mRNAs in migratory endothelial cells. We isolated RNA from endothelial cell protrusions from their cell bodies and used RNA-sequencing to identify asymmetrically distributed transcripts. These studies identified a set of transcripts enriched in endothelial cell protrusions over cell bodies, which included classically protrusion-enriched mRNAs and other intriguing transcripts encoding proteins implicated in cell migration. Overall design: To unveil the implications of mRNA localisation in endothelial cell migration during angiogenesis, Human Umbilical Vein Endothelial Cells (HUVECs) were cultured in modified Transwells that permit the segregation of protrusions formed at cell front from their cell body counterparts. In order to investigate whether HUVECs preferentially localise transcripts to their migratory front, RNA was extracted from cell bodies and protrusions to perform a genome-wide transcriptome screen (RNA-seq).