title: Alterations in insulin-signaling and coagulation pathways in platelets during hyperglycemia-hyperinsulinemia in healthy non-diabetic subject.
abstract: BACKGROUND
  Diabetes mellitus (DM) is a prothrombotic and proinflammatory state. Hyperglycemia (HG) is encountered even in patients without DM. We have shown that combined HG and hyperinsulinemia (HI) in healthy non-diabetic subjects increased circulating tissue factor (TF) and thrombin generation. To understand the changes in platelet and monocyte pathways induced by combined HG and HI in healthy non-diabetic state, we performed whole genome expression profiling of leukocyte-depleted platelets and monocytes before and after 24 hours of combined HG (glucose ~200mg/dL) and HI by glucose infusion clamp in a healthy non-diabetic subject.
  
  
  RESULTS
  We defined time-dependent differential mRNA expression (24 versus 0 hour fold change (FC) ≥ 2) common to platelets and monocytes. Ingenuity Pathways Analysis revealed alterations in canonical insulin receptor signaling and coagulation pathways. A preliminary group of 9 differentially expressed genes was selected for qRT-PCR confirmation. Platelet 24 hour sample was compared to the 0 hour sample plus 4 controls. Five transcripts in platelets and 6 in monocytes were confirmed. Platelet GSK3B and PTPN1 were upregulated, and STXBP4 was downregulated in insulin signaling, and F3 and TFPI were upregulated in coagulation pathways. Monocyte, PIK3C3, PTPN11 and TFPI were downregulated. Platelet GSKβ3 and PTPN11 protein and TF antigen in platelets and monocytes was increased.
  
  
  CONCLUSIONS
  Even in non-diabetic state, HG+HI for 24 hours induces changes in platelets and monocytes. They suggest downregulation of insulin signaling and upregulation of TF. Further studies are needed to elucidate cellular alterations leading to the prothrombotic and proinflammatory state in DM.
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA; Department of Medicine, Temple University School of Medicine, Philadelphia, PA, USA. Electronic address: koneti@temple.edu.
authors.forename: A Koneti
authors.initials: AK
authors.lastname: Rao
authors.affiliation: Division of Emergency Medicine, Children's National Medical Center, Washington, DC, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Freishtat
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA.
authors.forename: Gauthami
authors.initials: G
authors.lastname: Jalagadugula
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA.
authors.forename: Anamika
authors.initials: A
authors.lastname: Singh
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA.
authors.forename: Guangfen
authors.initials: G
authors.lastname: Mao
authors.affiliation: Division of Emergency Medicine, Children's National Medical Center, Washington, DC, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Wiles
authors.affiliation: General Clinical Research Center, Temple University School of Medicine, Philadelphia, PA, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Cheung
authors.affiliation: Department of Medicine, Temple University School of Medicine, Philadelphia, PA, USA; General Clinical Research Center, Temple University School of Medicine, Philadelphia, PA, USA.
authors.forename: Guenther
authors.initials: G
authors.lastname: Boden
mesh_terms: D000328:Adult; D001777:Blood Coagulation / Q000235:genetics; D001779:Blood Coagulation Factors / Q000235:genetics / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism*; D016000:Cluster Analysis; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D005819:Genetic Markers; D064368:Healthy Volunteers; D006801:Humans; D006943:Hyperglycemia / Q000097:blood* / Q000175:diagnosis / Q000235:genetics; D006946:Hyperinsulinism / Q000097:blood* / Q000175:diagnosis / Q000235:genetics; D007328:Insulin / Q000097:blood*; D008297:Male; D009000:Monocytes / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000097:blood; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics; D013997:Time Factors
chemical_list: D001779:Blood Coagulation Factors; D005819:Genetic Markers; D007328:Insulin; D012333:RNA, Messenger
keywords: Diabetes mellitus; Hyperglycemia-hyperinsulinemia; Insulin signaling; Monocytes; Platelets; Tissue factor