2026-04-17T06:47:40Zhttps://recercat.cat/oai/request

oai:recercat.cat:2072/4771322024-11-04T08:22:10Zcom_2072_98col_2072_378192

00925njm 22002777a 4500 dc Costales, Paula author Fuentes-Prior, Pablo author Castellano, José author Revuelta-López, Elena author Corral-Rodríguez, Maria Ángeles author Nasarre, Laura author Badimon, Lina author Llorente-Cortés, Vicenta author Universitat Autònoma de Barcelona author 2015 Low density lipoprotein receptor-related protein (LRP1) mediates the internalization of aggregated LDL (AgLDL), which in turn increases the expression of LRP1 in human vascular smooth muscle cells (hVSMCs). This positive feedback mechanism is thus highly efficient to promote the formation of hVSMC foam cells, a crucial vascular component determining the susceptibility of atherosclerotic plaque to rupture. Here we have determined the LRP1 domains involved in AgLDL recognition with the aim of specifically blocking AgLDL internalization in hVSMCs. The capacity of fluorescently labeled AgLDL to bind to functional LRP1 clusters was tested in a receptor-ligand fluorometric assay made by immobilizing soluble LRP1 "minireceptors"(sLRP1-II, sLRP1-III, and sLRP1-IV) recombinantly expressed in CHO cells. This assay showed that AgLDL binds to cluster II. We predicted three well exposed and potentially immunogenic peptides in the CR7-CR9 domains of this cluster (termed P1 (Cys-Glu), P2 (Asp-Cys), and P3 (Gly-Cys)). AgLDL, but not native LDL, bound specifically and tightly to P3-coated wells. Rabbit polyclonal antibodies raised against P3 prevented AgLDL uptake by hVSMCs and were almost twice as effective as anti-P1 and anti-P2 Abs in reducing intracellular cholesteryl ester accumulation. Moreover, anti-P3 Abs efficiently prevented AgLDL-induced LRP1 up-regulation and counteracted the down-regulatory effect of AgLDL on hVSMC migration. In conclusion, domain CR9 appears to be critical for LRP1-mediated AgLDL binding and internalization in hVSMCs. Our results open new avenues for an innovative anti-VSMC foam cell-based strategy for the treatment of vascular lipid deposition in atherosclerosis. http://hdl.handle.net/2072/477132 Amino Acid Sequence Animals Atherosclerotic plaque Cells, Cultured CHO Cells Cholesteryl esters Cricetinae Cricetulus Feedback mechanisms Fluorometric assay Foam Cells Human vascular smooth muscle cells Humans Ligands Lipoproteins, LDL Low density lipoprotein receptors Low Density Lipoprotein Receptor-Related Protein-1 Molecular Sequence Data Muscle, Smooth, Vascular Polyclonal antibody Real-Time Polymerase Chain Reaction Regulatory effects Sequence Homology, Amino Acid K domain CR9 of low density lipoprotein (LDL) receptor-related protein 1 (LRP1) is critical for aggregated LDL-induced foam cell formation from human vascular smooth muscle cells