title: GPR92/LPA₅ lysophosphatidate receptor mediates megakaryocytic cell shape change induced by human atherosclerotic plaques.
abstract: OBJECTIVE
  Oxidative processes and vascular inflammation underlying atherosclerosis lead to an accumulation of lysophosphatidic acid (LPA) molecules in the atheromatous intima. LPA, a platelet-activating component of human atherosclerotic plaques, possibly contributes to atherothrombus formation after plaque rupture. Human platelets express mRNA for the G protein-coupled receptors LPA₁₋₇ that derive from megakaryocytes. The aim of our study was to identify the functional LPA receptor(s) in human platelets by silencing individual LPA receptors in megakaryocytic (MK) cells.
  
  
  RESULTS
  We studied shape change of two human MK cell lines (Meg-01, Dami) by turbidometry, phase-contrast and scanning electron microscopy. They showed upon LPA stimulation a rapid, Rho-kinase-mediated shape change similar to that of human platelets. By qRT-PCR analysis we found expression of LPA₁₋₇ in both cell lines; LPA₄ and LPA₅ were the most abundant receptor transcripts. In both Meg-01 and Dami cells, the rank order of activation by LPA species was similar to that found in platelets: alkyl-LPA 18:1 > alkyl-LPA 16:0 > acyl-LPA 18:1 >> alkyl-LPA 18:0. Knock-down of individual LPA receptors by siRNA showed that LPA-mediated activation of MK cells was mediated by LPA₅, but not by LPA₁₋₄,₆,₇. Importantly, we found that human atherosclerotic plaque and lipid-rich core induced shape change of Dami cells, and that this effect was inhibited after LPA₅ silencing.
  
  
  CONCLUSIONS
  Our findings indicate that LPA₅ mediates LPA-induced shape change of MK cells and support its involvement in atherosclerotic plaque and lipid-rich core-mediated platelet activation. This receptor could be an attractive novel target for antithrombotic therapy.
authors.affiliation: Institute for Prevention of Cardiovascular Diseases, University of Munich, Munich, Germany.
authors.forename: Anna L
authors.initials: AL
authors.lastname: Khandoga
authors.forename: Dharmendra
authors.initials: D
authors.lastname: Pandey
authors.forename: Ulrich
authors.initials: U
authors.lastname: Welsch
authors.forename: Richard
authors.initials: R
authors.lastname: Brandl
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Siess
mesh_terms: D050197:Atherosclerosis / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D048430:Cell Shape; D006801:Humans; D050356:Lipid Metabolism; D008246:Lysophospholipids / Q000378:metabolism*; D008533:Megakaryocytes / Q000378:metabolism* / Q000473:pathology; D008855:Microscopy, Electron, Scanning; D009391:Nephelometry and Turbidimetry; D034622:RNA Interference; D049368:Receptors, Lysophosphatidic Acid / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: C512992:LPAR5 protein, human; D008246:Lysophospholipids; D049368:Receptors, Lysophosphatidic Acid; D054460:rho-Associated Kinases; C032881:lysophosphatidic acid