Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma

Abstract

The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). A total of 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls were recruited in this study. DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. Moreover, serum sialic acid (SA) level was measured by enzymatic method. We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. The sensitivity and specificity of Peak3 were 88.1% and 92.9%, while Peak7 were 92.9% and 97.6%, respectively. The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. Serum SA level was significantly higher in patients with LCMM than that in healthy controls. Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM.

Capillary electrophoresis analysis of N‐glycosylation changes of serum paraproteins in multiple myeloma

“…confirmed that reduced homing and engraftment of ST3GAL6 sialyltransferase knockdown in MM cells in the bone marrow decreased tumor propagation . Decreased core fucosylation, appearance of bisecting forms and increased sialylation of serum glycoproteins were observed in patients with light‐chain multiple myeloma (LCMM) . Also in MM, recently low galactosylation and fucosylation was found .…”

Serum N‐glycans outperform CA19‐9 in diagnosis of extrahepatic cholangiocarcinoma

“…However, the DSA‐FACE method remains an effective glycomic approach for the discovery of cancer biomarkers. Differential biomarkers of clinical application potential can be identified with simple structural information using this method with higher repeatability and better quality control than the original method, as previously reported . Aberrant sialylation has been implicated in several cancers .…”

“…However, most of these approaches involve chromatographic separation, mass spectrometry analysis or various microarrays, along with high complexity and low cost‐efficiency . Herein, we employ a modified DNA sequencer‐assisted fluorophore‐assisted capillary electrophoresis (DSA‐FACE) technology, which is originally established by Callewaert and coworkers to decipher the global N ‐glycome of serum . Discovery of differentially expressed glycans from whole serum may serve to present candidate biomarkers for the discrimination of ECCA.…”

Capillary Electrophoresis Separations of Glycans

“…The benefit of modifying N -glycan samples prior to CE separations using enzymes was recognized early. 90 Applications with CE have been performed off-line with a variety of enzymes 28 , 59 , 88 , 91 and typically focus on removal of sialic acids 52 , 66 , 92 in some cases as a means to reduce complexity of the CE separation. 92 These reactions are typically performed overnight at an elevated temperature and a specified pH.…”