Identification of VE-cadherin associated proteins regulated through the lncRNA LASSIE

The shear stress-regulated lncRNA LASSIE interacts with cytoskeletal proteins as shown by RNA-antisense purification and subsequent mass spectrometry analysis using an anti-LASSIE and a non-targeting 3’ Desthiobiotin-TEG labeled 2’ O-Me-RNA. As this analysis resulted in high protein binding to the non-targeting control oligonucleotide, the RNA antisense purification was repeated in HUVECs treated with control and anti-LASSIE siRNA. Mass spectrometry analysis confirmed binding of LASSIE to the Intermediate filament protein nestin, as nestin enrichment was decreased in the absence of LASSIE. Additional functional analyses demonstrate a role of LASSIE in shear stress sensing and barrier function in endothelial cells, by stabilizing endothelial cell junctions through connection to the cytoskeleton.