Proteomic analysis of maternal serum in down syndrome: identification of novel protein biomarkers.

Down syndrome (DS) is the most prevalent chromosomal disorder, accounting for significant morbidity and mortality. Definitive diagnosis requires invasive amniocentesis, and current maternal serum-based testing requires a false-positive rate of about 5% to detect 85% of affected pregnancies. We have performed a comprehensive proteomic analysis to identify potential serum biomarkers to detect DS. First- and second-trimester maternal serum samples of DS and gestational age-matched controls were analyzed using multiple, complementary proteomic approaches, including fluorescence 2-dimensional gel electrophoresis (2D-DIGE), 2-dimensional liquid chromatography-chromatofocusing (2D-CF), multidimensional protein identification technology (MudPIT; LC/LC-MS/MS), and MALDI-TOF-MS peptide profiling. In total, 28 and 26 proteins were differentially present in first- and second-trimester samples, respectively. Of these, 19 were specific for the first trimester and 16 for the second trimester, and 10 were differentially present in both trimesters. Analysis of MALDI-TOF-MS peptide profiles with pattern-recognition software also discriminated between DS and controls in both trimesters, with an average recognition capability approaching 96%. A majority of the biomarkers identified are serum glycoproteins that may play a role in cellular differentiation and growth of fetus. Further characterization and quantification of these markers in a larger cohort of subjects may provide the basis for new tests for improved DS screening.

[1]  Walter E. Johnson,et al.  An association between low maternal serum α-fetoprotein and fetal chromosomal abnormalities , 1984 .

[2]  Leigh Anderson,et al.  Candidate‐based proteomics in the search for biomarkers of cardiovascular disease , 2005, The Journal of physiology.

[3]  Arul M Chinnaiyan,et al.  Protein microarrays using liquid phase fractionation of cell lysates , 2003, Proteomics.

[4]  F. Malone,et al.  Pregnancy Loss Rates After Midtrimester Amniocentesis , 2006, Obstetrics and gynecology.

[5]  G. Avvakumov,et al.  Pregnancy-associated molecular variants of human serum transcortin and thyroxine-binding globulin. , 1984, Carbohydrate research.

[6]  Michael K. Coleman,et al.  Correlation of relative abundance ratios derived from peptide ion chromatograms and spectrum counting for quantitative proteomic analysis using stable isotope labeling. , 2005, Analytical chemistry.

[7]  L L Needham,et al.  Isotope dilution--mass spectrometric quantification of specific proteins: model application with apolipoprotein A-I. , 1996, Clinical chemistry.

[8]  Lora J. H. Bean,et al.  Epidemiology of Down syndrome. , 2007, Mental retardation and developmental disabilities research reviews.

[9]  K. Resing,et al.  Comparison of Label-free Methods for Quantifying Human Proteins by Shotgun Proteomics*S , 2005, Molecular & Cellular Proteomics.

[10]  B. Searle,et al.  Identification of protein modifications using MS/MS de novo sequencing and the OpenSea alignment algorithm. , 2005, Journal of proteome research.

[11]  G. Barkai,et al.  Analysis of fetal blood cells in the maternal circulation: challenges, ongoing efforts, and potential solutions. , 2004, Stem cells and development.

[12]  Robertson Craig,et al.  TANDEM: matching proteins with tandem mass spectra. , 2004, Bioinformatics.

[13]  B. Searle,et al.  High-throughput identification of proteins and unanticipated sequence modifications using a mass-based alignment algorithm for MS/MS de novo sequencing results. , 2004, Analytical chemistry.

[14]  J. Palo,et al.  Serum amysoid A protein, albumin and prealbumin in Alzheimer's disease and in demented patients with Down's syndrome , 1986 .

[15]  P. Lansbury,et al.  Amyloid diseases: abnormal protein aggregation in neurodegeneration. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Raynes.  Variations in the relative proportions of microheterogeneous forms of plasma glycoproteins in pregnancy and disease. , 1982, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[17]  R. D. Wilson.  Cell-free fetal DNA in the maternal circulation and its future uses in obstetrics. , 2005, Journal of obstetrics and gynaecology Canada : JOGC = Journal d'obstetrique et gynecologie du Canada : JOGC.

[18]  J. Richie,et al.  Chromatofocusing fractionation and two‐dimensional difference gel electrophoresis for low abundance serum proteins , 2005, Proteomics.

[19]  A. Piñeiro,et al.  ITIH4 serum concentration increases during acute-phase processes in human patients and is up-regulated by interleukin-6 in hepatocarcinoma HepG2 cells. , 1999, Biochemical and biophysical research communications.

[20]  D. Lubman,et al.  Two-dimensional liquid chromatography protein expression mapping for differential proteomic analysis of normal and O157:H7 Escherichia coli. , 2003, BioTechniques.

[21]  J. Egan,et al.  Down syndrome births in the United States from 1989 to 2001. , 2004, American journal of obstetrics and gynecology.

[22]  N J Wald,et al.  First and second trimester antenatal screening for Down's syndrome: the results of the Serum, Urine and Ultrasound Screening Study (SURUSS). , 2003, Health technology assessment.

[23]  F. R. Brown,et al.  Immunologic dysfunction: simultaneous study of alzheimer's and older down's patients , 1987, Mechanisms of Ageing and Development.

[24]  L. Mucke,et al.  Modulation of Alzheimer-Like Synaptic and Cholinergic Deficits in Transgenic Mice by Human Apolipoprotein E Depends on Isoform , Aging, and Overexpression of Amyloid β Peptides But Not on Plaque Formation , 2002, The Journal of Neuroscience.

[25]  J. Lapidus,et al.  Diagnosis of intra-amniotic infection by proteomic profiling and identification of novel biomarkers. , 2004, JAMA.

[26]  U. Hellman,et al.  Amyloid deposits in transthyretin‐derived amyloidosis: cleaved transthyretin is associated with distinct amyloid morphology , 2005, The Journal of pathology.

[27]  Y. Lo.  Recent advances in fetal nucleic acids in maternal plasma. , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[28]  Vanathi Gopalakrishnan,et al.  Proteomic profiling of cerebrospinal fluid identifies biomarkers for amyotrophic lateral sclerosis , 2005, Journal of neurochemistry.

[29]  D. Selkoe,et al.  Temporal accrual of complement proteins in amyloid plaques in Down's syndrome with Alzheimer's disease. , 2000, The American journal of pathology.

[30]  D. Torpy,et al.  Pregnancy-associated corticosteroid-binding globulin: high resolution separation of glycan isoforms. , 2004, Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme.

[31]  A. Caughey,et al.  Ultrasound screening of fetuses at increased risk for Down syndrome: how many missed diagnoses? , 2006, Prenatal diagnosis.

[32]  U. Reddy,et al.  Incorporating first-trimester Down syndrome studies into prenatal screening: executive summary of the National Institute of Child Health and Human Development workshop. , 2006, Obstetrics and gynecology.

[33]  Julie Perkins,et al.  Proteomic analysis of human serum by two-dimensional differential gel electrophoresis after depletion of high-abundant proteins. , 2004, Journal of proteome research.

[34]  M. Albert,et al.  Genetic association of an (α2-macroglobulin (Val1000lle) polymorphism and Alzheimer's disease , 1998 .

[35]  L. Cole,et al.  Sialic acid–deficient invasive trophoblast antigen (sd‐ITA): a new urinary variant for gestational Down syndrome screening , 2004, Prenatal diagnosis.

[36]  W. Borth.  α2“Macroglobulin, a multifunctional binding protein with targeting characteristics , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.