The goal of this study was to explore the plasma proteome of myelodysplastic syndrome (MDS) patients with refractory anemia with excess blasts subtype 2 (RAEB-2) in comparison to healthy controls. 20 plasma samples were separated with 2D electrophoresis and statistically processed with Progenesis SameSpots software. 47 significantly differing (P < 0.05) spots were observed, and 27 different proteins were identified by nano-LC-MS/MS. Mass spectrometry-based relative label-free quantification showed a 2-fold increase of the leucine-rich alpha-2-glycoprotein (LRAG) peptide levels in the RAEB-2 group. Changes in the fragments of the inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) protein were observed. Western blot analysis showed no differences in albumin and ITIH4 levels, while increased expression was observed for LRAG in the RAEB-2 group. Quantification using ELISA showed decreased plasma level of alpha-2-HS glycoprotein in the RAEB-2 group. In conclusion, this is the first time that alpha-2-HS glycoprotein and LRAG were proposed as new biomarkers of RAEB-2 and advanced MDS, respectively. Alpha-2-HS glycoprotein, a protein involved in the bone marrow development and previously proposed as a MDS biomarker candidate, was significantly decreased in RAEB-2. Increased expression and changes in modification(s) were observed for LRAG, a protein involved in granulocytic and neutrophil differentiation, and angiogenesis.

Institute of Hematology and Blood Transfusion U Nemocnice 1 128 20 Prague 2 Czech Republic

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Belickova M, Merkerova MD, Stara E, et al. DNA repair gene variants are associated with an increased risk of myelodysplastic syndromes in a Czech population. Journal of Hematology and Oncology. 2013;6(1, article 9) PubMed PMC

Merkerova MD, Bystricka D, Belickova M, et al. From cryptic chromosomal lesions to pathologically relevant genes: integration of SNP-array with gene expression profiling in myelodysplastic syndrome with normal karyotype. Genes Chromosomes and Cancer. 2012;51(5):419–428. PubMed

Cechova H, Lassuthova P, Novakova L, et al. Monitoring of methylation changes in 9p21 region in patients with myelodysplastic syndromes and acute myeloid leukemia. Neoplasma. 2012;59(2):168–174. PubMed

Raza A, Galili N. The genetic basis of phenotypic heterogeneity in myelodysplastic syndromes. Nature Reviews Cancer. 2012;12(12):849–859. PubMed

Rodriguez-Emmenegger C, Houska M, Alles AB, Brynda E. Surfaces resistant to fouling from biological fluids: towards bioactive surfaces for real applications. Macromolecular Bioscience. 2012;12(10):1413–1422. PubMed

Riedel T, Brynda E, Dyr JE, Houska M. Controlled preparation of thin fibrin films immobilized at solid surfaces. Journal of Biomedical Materials Research Part A. 2009;88(2):437–447. PubMed

Emmenegger CR, Brynda E, Riedel T, Sedlakova Z, Houska M, Alles AB. Interaction of blood plasma with antifouling surfaces. Langmuir. 2009;25(11):6328–6333. PubMed

Riedel T, Riedelová-Reicheltová Z, Májek P, et al. Complete identification of proteins responsible for human blood plasma fouling on poly(ethylene glycol)-based surfaces. Langmuir. 2013;29(10):3388–3397. PubMed

Májek P, Riedelová-Reicheltová Z, Suttnar J, Pečánková K, Čermák J, Dyr JE. Plasma proteome changes associated with refractory anemia and refractory anemia with ringed sideroblasts in patients with myelodysplastic syndrome. Proteome Science. 2013;11(1, article 14) PubMed PMC

Májek P, Reicheltová Z, Suttnar J, Čermák J, Dyr JE. Plasma proteome changes associated with refractory cytopenia with multilineage dysplasia. Proteome Science. 2011;9, article 64 PubMed PMC

Májek P, Reicheltová Z, Suttnar J, Čermák J, Dyr JE. Plasma protein alterations in the refractory anemia with excess blasts subtype 1 subgroup of myelodysplastic syndrome. Proteome Science. 2012;10(1, article 31) PubMed PMC

Malcovati L, Germing U, Kuendgen A, et al. Time-dependent prognostic scoring system for predicting survival and leukemic evolution in myelodysplastic syndromes. Journal of Clinical Oncology. 2007;25(23):3503–3510. PubMed

Vardiman JW, Harris NL, Brunning RD. The World Health Organization (WHO) classification of the myeloid neoplasms. Blood. 2002;100(7):2292–2302. PubMed

Májek P, Reicheltová Z, Štikarová J, Suttnar J, Sobotková A, Dyr JE. Proteome changes in platelets activated by arachidonic acid, collagen, and thrombin. Proteome Science. 2010;8, article 56 PubMed PMC

Lange V, Picotti P, Domon B, Aebersold R. Selected reaction monitoring for quantitative proteomics: a tutorial. Molecular Systems Biology. 2008;4, article 222 PubMed PMC

Kay R, Barton C, Ratcliffe L, et al. Enrichment of low molecular weight serum proteins using acetonitrile precipitation for mass spectrometry based proteomic analysis. Rapid Communications in Mass Spectrometry. 2008;22(20):3255–3260. PubMed

Májek P, Reicheltová Z, Suttnar J, et al. Plasma proteome changes in cardiovascular disease patients: novel isoforms of apolipoprotein A1. Journal of Translational Medicine. 2011;9, article 84 PubMed PMC

Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nature Methods. 2012;9(7):671–675. PubMed PMC

Ma W, Kantarjian H, Bekele B, et al. Proteasome enzymatic activities in plasma as risk stratification of patients with acute myeloid leukemia and advanced-stage myelodysplastic syndrome. Clinical Cancer Research. 2009;15(11):3820–3826. PubMed PMC

Abdullah-Soheimi SS, Lim B-K, Hashim OH, Shuib AS. Patients with ovarian carcinoma excrete different altered levels of urine CD59, kininogen-1 and fragments of inter-alpha-trypsin inhibitor heavy chain H4 and albumin. Proteome Science. 2010;8, article 58 PubMed PMC

Yoshida Y, Takahashi Y, Yoshikawa T, Nonomura A, Yoshioka A. Suppressive effect of α2 Heremans-Schmid glycoprotein on in vitro calcification of osteogenesis. Pediatrics International. 2006;48(1):11–16. PubMed

Lebreton JP, Joisel F, Raoult JP, Lannuzel B, Rogez JP, Humbert G. Serum concentration of human alpha 2 HS glycoprotein during the inflammatory process. Evidence that alpha 2 HS glycoprotein is a negative acute-phase reactant. Journal of Clinical Investigation. 1979;64(4):1118–1129. PubMed PMC

Daveau M, Davrinche C, Julen N, Hiron M, Arnaud P, Lebreton J-P. The synthesis of human α-2-HS glycoprotein is down-regulated by cytokines in hepatoma HepG2 cells. FEBS Letters. 1988;241(1-2):191–194. PubMed

Petrik V, Saadoun S, Loosemore A, et al. Serum α2-HS glycoprotein predicts survival in patients with glioblastoma. Clinical Chemistry. 2008;54(4):713–722. PubMed

O’Donnell LC, Druhan LJ, Avalos BR. Molecular characterization and expression analysis of leucine-rich α2-glycoprotein, a novel marker of granulocytic differentiation. Journal of Leukocyte Biology. 2002;72(3):478–485. PubMed

Serada S, Fujimoto M, Terabe F, et al. Serum leucine-rich alpha-2 glycoprotein is a disease activity biomarker in ulcerative colitis. Inflammatory Bowel Diseases. 2012;18(11):2169–2179. PubMed

Gazouli M, Anagnostopoulos AK, Papadopoulou A, et al. Serum protein profile of Crohn’s disease treated with infliximab. Journal of Crohn’s and Colitis. 2013;7:e461–e470. PubMed

Liu Y, Luo X, Hu H, et al. Integrative Proteomics and Tissue Microarray Profiling Indicate the Association between Overexpressed Serum Proteins and Non-Small Cell Lung Cancer. PLoS ONE. 2012;7(12)e51748 PubMed PMC

Kentsis A, Ahmed S, Kurek K, et al. Detection and diagnostic value of urine leucine-rich α-2-glycoprotein in children with suspected acute appendicitis. Annals of Emergency Medicine. 2012;60(1):78–83. PubMed PMC

Wang X, Abraham S, McKenzie JAG, et al. LRG1 promotes angiogenesis by modulating endothelial TGF-β signalling. Nature. 2013;499(7458):306–311. PubMed PMC

Riedel T, Rodriguez-Emmenegger C, de los Santos Pereira A, et al. Diagnosis of Epstein-Barr virus infection in clinical serum samples by an SPR biosensor assay. Biosensors and Bioelectronics. 2014;55:278–284. PubMed

Rodriguez-Emmenegger C, Hasan E, Pop-Georgievski O, Houska M, Brynda E, Alles AB. Controlled/living surface-initiated ATRP of antifouling polymer brushes from gold in PBS and blood sera as a model study for polymer modifications in complex biological media. Macromolecular Bioscience. 2012;12(4):525–532. PubMed

Mariani S, Minunni M. Surface plasmon resonance applications in clinical analysis. Analytical and Bioanalytical Chemistry. 2014;406:2303–2323. PubMed PMC

McGrath TF, Buijs J, Huet AC, Delahaut P, Elliott CT, Mooney MH. Assessment of a multiplexing high throughput immunochemical SPR biosensor in measuring multiple proteins on a single biosensor chip. Sensors and Actuators B. 2013;186:423–430.

Fracchiolla NS, Artuso S, Cortelezzi A. Biosensors in clinical practice: focus on oncohematology. Sensors. 2013;13(5):6423–6447. PubMed PMC

Proteomic Case Studies of MDS in Progression: Heterogeneity and More Heterogeneity

Plasma Protein Biomarker Candidates for Myelodysplastic Syndrome Subgroups

Peripheral blood mononuclear cell proteome changes in patients with myelodysplastic syndrome