Plasma proteome changes associated with refractory anemia and refractory anemia with ringed sideroblasts in patients with myelodysplastic syndrome

Abstract

BackgroundRefractory anemia and refractory anemia with ringed sideroblasts are two myelodysplastic syndrome (MDS) subgroups linked with anemia. MDS is a group of heterogeneous oncohematological bone marrow disorders characterized by ineffective hematopoiesis, blood cytopenias, and progression of the disease toward acute myeloid leukemia. The aim of this study was to search for plasma proteome changes in MDS patients with refractory anemia and refractory anemia with ringed sideroblasts.ResultsA total of 26 patient and healthy donor plasma samples were depleted of fourteen high-abundant plasma proteins, separated with 2D electrophoresis, and statistically processed with Progenesis SameSpots software. 55 significantly differing spots were observed and corresponded to 39 different proteins identified by nanoLC-MS/MS. Changes in the fragments of the inter-alpha-trypsin inhibitor heavy chain H4 protein were observed. Using mass spectrometry-based relative label-free quantification of tryptic peptides, there were differences in alpha-2-HS-glycoprotein peptides, while no differences were observed between the control and patient sample groups for retinol-binding protein 4 peptides.ConclusionsThis study describes plasma proteome changes associated with MDS patients with refractory anemia and refractory anemia with ringed sideroblasts. Changes observed in the inter-alpha-trypsin inhibitor heavy chain H4 fragments were in agreement with our previous studies of other MDS subgroups: refractory cytopenia with multilineage dysplasia and refractory anemia with excess blasts subtype 1. Mass spectrometry-based relative quantification of retinol-binding protein 4 peptides has shown that there are differences in the modification of this protein between refractory anemia with excess blasts subtype 1 patients and MDS patients with refractory anemia and refractory anemia with ringed sideroblasts. Alpha-2-HS-glycoprotein seems to be a new potential MDS biomarker candidate.

Proteome Changes in the Plasma of Myelodysplastic Syndrome Patients with Refractory Anemia with Excess Blasts Subtype 2

“…Contrary to previous studies of low- or intermediate-risk MDS subgroups, this study interrogates the high-risk subgroup RAEB-2 and thus has potential to find protein markers highly valuable for monitoring the disease development. Comparing the overall results with respect to the number of changes (spots) that were identified in all four studies, it is obvious that there is no correlation (numbers of spots) with WPSS score, as has been pointed out previously [9]. Again, it seems that “a depth of changes” is the principal factor; however, the limitations of the methods used have to be considered, especially the limited level of protein detection.…”

Peripheral Blood Mononuclear Cell Proteome Changes in Patients with Myelodysplastic Syndrome

“…Isoelectric focusing was performed (IPG strips pI 4–7, 7.7 cm) followed by SDS-PAGE (8 × 10 cm, 10% resolving gel, 3.75% stacking gel, and 30 mA/gel), as described in detail in previous publications [ 4 – 6 , 22 ]. Briefly, 40 μ g of cytosolic, 50 μ g of membrane and membrane organelle, and 40 μ g of nuclear proteins were used for an IPG strip.…”

“…After each staining step, the gels were digitized and processed with Progenesis SameSpots software (Nonlinear Dynamics, Newcastle upon Tyne, UK) that computed the fold and P values of all spots using one-way ANOVA analysis. Protein spots that differed significantly ( P < 0.05) were submitted for protein identification by tandem mass spectrometry (HCT ultra ion-trap mass spectrometer with nanoelectrospray ionization; Bruker Daltonics, Bremen, Germany) coupled to a nano-LC system (UltiMate 3000; Dionex, Sunnyvale, CA, USA); this procedure has been described in detail previously [ 4 – 6 , 22 ]. Mascot (Matrix Science, London, UK) was used for database searching (Swiss-Prot).…”

“…In recent years, fundamental knowledge in MDS pathophysiology based on DNA alterations has been and is still being complemented by other “ omics ” disciplines, in particular by proteomics, with the proteomes of plasma (of different MDS subgroups such as refractory anemia and refractory anemia with ringed sideroblasts (RA and RARS) [ 4 ], RCMD [ 5 ], refractory anemia with excess of blasts subtype 1 (RAEB-1) [ 6 ], and refractory anemia with excess of blasts subtype 2 (RAEB-2) [ 7 ]), serum [ 8 , 9 ], platelets [ 10 ], and neutrophils [ 11 ] of MDS patients having been investigated. Despite research efforts the pathogenesis and exact mechanisms of MDS development still remain unclear, as there are many factors involved.…”

Surface Plasmon Resonance: Advances of Label-Free Approaches in The Analysis of Biological Samples

“…More recent reports presented the detection of fetuin A and squamous cell carcinoma antigen [60,61]. The former was shown to be a biomarker of myelodysplastic syndrome [62]. Zhao et al presented a novel and reusable biosensor based on LSPR for the detection of squamous cell carcinoma antigen in human serum [50].…”