Discovery and validation of serum protein changes in type 1 diabetes patients using high throughput two dimensional liquid chromatography-mass spectrometry and immunoassays

Mol Cell Proteomics. 2011 Nov;10(11):M111.012203. doi: 10.1074/mcp.M111.012203. Epub 2011 Sep 6.

Abstract

Type 1 diabetes (T1D) is expected to cause significant changes in the serum proteome; however, few studies have systematically assessed the proteomic profile change associated with the disease. In this study, a semiquantitative spectral counting-based two dimensional liquid chromatography mass spectrometry platform was used to analyze serum samples from T1D patients and controls. In this discovery phase, significant differences were found for 21 serum proteins implicated in inflammation, oxidation, metabolic regulation, and autoimmunity. To assess the validity of these findings, six candidate proteins including adiponectin, insulin-like growth factor binding protein 2, serum amyloid protein A, C-reactive protein, myeloperoxidase, and transforming growth factor beta induced were selected for subsequent immune assays for 1139 T1D patients and 848 controls. A series of statistical analyses using cases and controls matched for age, sex, and genetic risk confirmed that T1D patients have significantly higher serum levels for four of the six proteins: adiponectin (odds ratio (OR) = 1.95, p = 10(-27)), insulin-like growth factor binding protein 2 (OR = 2.02, p < 10(-20)), C-reactive protein (OR = 1.13, p = 0.007), serum amyloid protein A (OR = 1.51, p < 10(-16)); whereas the serum levels were significantly lower in patients than controls for the two other proteins: transforming growth factor beta induced (OR = 0.74, p < 10(-5)) and myeloperoxidase (OR = 0.51, p < 10(-41)). Compared with subjects in the bottom quartile, subjects in the top quartile for adiponectin (OR = 6.29, p < 10(-37)), insulin-like growth factor binding protein 2 (OR = 7.95, p < 10(-46)), C-reactive protein (OR = 1.38, p = 0.025), serum amyloid protein A (OR = 3.36, p < 10(-16)) had the highest risk of T1D, whereas subjects in the top quartile of transforming growth factor beta induced (OR = 0.41, p < 10(-11)) and myeloperoxidase (OR = 0.10, p < 10(-43)) had the lowest risk of T1D. These findings provided valuable information on the proteomic changes in the sera of T1D patients.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adiponectin / blood*
  • Adolescent
  • Adult
  • Age Factors
  • Aged
  • Aged, 80 and over
  • Biomarkers / blood
  • C-Reactive Protein / metabolism*
  • Case-Control Studies
  • Child
  • Child, Preschool
  • Chromatography, Liquid
  • Diabetes Mellitus, Type 1 / blood*
  • Enzyme-Linked Immunosorbent Assay
  • Extracellular Matrix Proteins / blood
  • Female
  • Humans
  • Infant
  • Infant, Newborn
  • Insulin-Like Growth Factor Binding Protein 2 / blood*
  • Male
  • Middle Aged
  • Multivariate Analysis
  • Peroxidase / blood
  • Serum Amyloid A Protein / metabolism*
  • Sex Factors
  • Tandem Mass Spectrometry
  • Transforming Growth Factor beta / blood
  • Young Adult

Substances

  • Adiponectin
  • Biomarkers
  • Extracellular Matrix Proteins
  • Insulin-Like Growth Factor Binding Protein 2
  • Serum Amyloid A Protein
  • Transforming Growth Factor beta
  • betaIG-H3 protein
  • C-Reactive Protein
  • Peroxidase