title: TRIB3 R84 variant is associated with impaired insulin-mediated nitric oxide production in human endothelial cells.
abstract: BACKGROUND
  In the endothelium, insulin promotes nitric oxide (NO) production, through the insulin receptor/IRS-1/PI3-Kinase/Akt/eNOS signaling pathway. An inhibitor of insulin action, TRIB3, has recently been identified which affects insulin action by binding to and inhibiting Akt phosphorylation. We have recently described a Q84R gain-of-function polymorphism of TRIB3 with the R84 variant being associated with insulin resistance and an earlier age at myocardial infarction.
  
  
  RESULTS
  To investigate the TRIB3 R84 variant impact on endothelial insulin action, we cultured human umbilical vein endothelial cells (HUVECs) naturally carrying different TRIB3 genotypes (QQ-, QR-, or RR-HUVECs). TRIB3 inhibitory activity on insulin-stimulated Akt phosphorylation and the amount of protein which was coimmunoprecipitable with Akt were significantly greater in QR- and RR- as compared to QQ- HUVECs. After insulin stimulation, Akt and eNOS activation as well as NO production were markedly decreased in QR- and RR- as compared to QQ-HUVECs. TRIB3 molecular modeling analysis provided insights into the structural changes related to the polymorphisms potentially determining differences in protein-protein interaction with Akt.
  
  
  CONCLUSIONS
  Our data demonstrate that the TRIB3 R84 variant impairs insulin signaling and NO production in human endothelial cells. This finding provides a plausible biological background for the deleterious role of TRIB3 R84 on genetic susceptibility to coronary artery disease.
authors.affiliation: Department of Medicine and Aging Sciences, Edificio CeSi, room 271, University of Chieti, via dei Vestini 1, 66100 CHIETI, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Andreozzi
authors.forename: Gloria
authors.initials: G
authors.lastname: Formoso
authors.forename: Sabrina
authors.initials: S
authors.lastname: Prudente
authors.forename: Marta Letizia
authors.initials: ML
authors.lastname: Hribal
authors.forename: Assunta
authors.initials: A
authors.lastname: Pandolfi
authors.forename: Emanuele
authors.initials: E
authors.lastname: Bellacchio
authors.forename: Sara
authors.initials: S
authors.lastname: Di Silvestre
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Trischitta
authors.forename: Agostino
authors.initials: A
authors.lastname: Consoli
authors.forename: Giorgio
authors.initials: G
authors.lastname: Sesti
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D001665:Binding Sites; D002318:Cardiovascular Diseases / Q000378:metabolism / Q000503:physiopathology; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D042783:Endothelial Cells / Q000201:enzymology / Q000378:metabolism*; D004789:Enzyme Activation; D005838:Genotype; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D055504:Insulin Receptor Substrate Proteins; D007333:Insulin Resistance / Q000235:genetics; D008958:Models, Molecular; D009154:Mutation; D009569:Nitric Oxide / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D051058:Oncogene Protein v-akt / Q000378:metabolism; D010641:Phenotype; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000037:antagonists & inhibitors; D011972:Receptor, Insulin / Q000378:metabolism; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D006152:Cyclic GMP; D038362:Glycogen Synthase Kinase 3; D007328:Insulin; D055504:Insulin Receptor Substrate Proteins; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D051058:Oncogene Protein v-akt; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D011972:Receptor, Insulin; D012097:Repressor Proteins; C526292:IRS1 protein, human; C474831:TRIB3 protein, human; C496314:NOS3 protein, human