title: Folate deficiency induced hyperhomocysteinemia changes the expression of thrombosis-related genes.
abstract: Hyperhomocysteinemia (HH) is an independent risk factor for thrombosis although the precise pathogenesis is still unresolved. Previous studies have demonstrated that HH changes whole blood coagulation by increasing the velocity, increasing the firmness of the formed clot, and by prolonging the initiation phase of the coagulation. With the aim of elucidating the genetic pathogenesis which might be responsible for the changes in whole blood coagulation, we applied oligo-array technology to RNA from buffycoat-cells comparing animals suffering from hyperhomocysteinemia (42 micromol/l) with controls (6 micromol/l). Data mining identified a number of relevant genes, and the expression pattern was validated by real time reverse transcriptase-polymerase chain reaction. An upregulation of integrin beta-3, Rap 1b, glycoprotein V, platelet-endothelial cell adhesion molecule-1 (PECAM-1) and von Willebrand factor (vWF) led us to deduce increased platelet activation/aggregation. Coagulation factor XIIIa was upregulated and may contribute in increasing the firmness of the formed clot. Impaired fibrinolysis was anticipated, since an upregulation of plasminogen activator inhibitor-1 (PAI-1) and a downregulation of tissue-type plasminogen activator (t-PA) were detected. Reduced spontaneous contact activation was anticipated due to a downregulation of the kallikrein gene. Upregulation of selectins may contribute to increased tethering and rolling of leukocytes. In conclusion, folate deficiency induced hyperhomocysteinemia changes in the gene expression of buffy coat cells which was characterized by increased platelet activation, impaired fibrinolysis and a reduced contact activation of the coagulation. These changes may contribute to explain the increased risk of thrombosis seen in hyperhomocysteinemia individuals. This pattern of the hyperhomocysteinemia-affected genes may represent a reference for further studies at the protein level to define the folate depletion effects in blood cells.
authors.affiliation: Clinical Institute, University Hospital of Aarhus, Skejby Sygehus, Brendstrupgaardsvej, Denmark. lse@dadlnet.dk
authors.forename: Liselotte S
authors.initials: LS
authors.lastname: Ebbesen
authors.forename: Sanne H
authors.initials: SH
authors.lastname: Olesen
authors.forename: Mogens
authors.initials: M
authors.lastname: Kruhøffer
authors.forename: Jørgen
authors.initials: J
authors.lastname: Ingerslev
authors.forename: Torben F
authors.initials: TF
authors.lastname: Ørntoft
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005494:Folic Acid Deficiency / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D020138:Hyperhomocysteinemia / Q000378:metabolism*; D039661:Integrin beta3 / Q000235:genetics; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013927:Thrombosis / Q000235:genetics* / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000235:genetics; D020668:rap GTP-Binding Proteins / Q000235:genetics; D014841:von Willebrand Factor / Q000235:genetics
chemical_list: D039661:Integrin beta3; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D019038:Platelet Glycoprotein GPIb-IX Complex; D014841:von Willebrand Factor; D010959:Tissue Plasminogen Activator; D020668:rap GTP-Binding Proteins