Klin Padiatr 2014; 226 - O_02
DOI: 10.1055/s-0034-1371098

Glycoproteomic Profiling of Hodgkin Lymphoma Reveals Novel Proteins that Contribute to the Microenvironment and Have Diagnostic Utility

M Lim 1, D Rolland 1, N Brown 1, YK Jeon 2, K Elenitoba-Johnson 1
  • 1University of Michigan, Pathology, Ann Arbor, United States
  • 2Seoul National University, Seoul, Republic of Korea

Background: Proteins with N-linked glycosylation are frequently expressed on cell membranes and/or secreted, and represent idea biomarkers with diagnostic and therapeutic utility. In this study, we carried out unbiased identification of N-linked glycoproteins of Hodgkin lymphoma-derived cell lines.

Design/Methods: Glycoproteomic analysis was carried out for 3 classical Hodgkin lymphoma (CHL) cell lines using solid-phase extraction of N-linked glycopeptides (SPEG) followed by liquid chromatography/tandem mass spectrometry. The expression of a subset of proteins was validated using western blot and immunohistochemistry on patient-derived biopsies.

Results: In total we identified 510 glycoproteins from 3 cell lines representative of classical Hodgkin lymphoma. Unsupervised hierarchical clustering of the glycoproteins successfully classified the cell lines distinct from B-and T- cell non-Hodgkin lymphoma-derived cell lines. Previously known CD proteins and cytokine receptors expressed in CHL (i.e., CD30, CD40, IL13R) were identified. Notably, novel proteins that regulate tumor microenvironment (cytokine receptors, receptor tyrosine kinases, growth factors and cell adhesion molecules) were identified in one or more CHL cell lines. Evaluation of ICAM-1, a carrier protein of the CD15 epitope in 203 CHL tissue biopsies using immunohistochemistry demonstrated a sensitivity of 91.1% in identifying Reed-Sternberg cells which was significantly higher than that of CD15 (62.1%).

Conclusion: Our mass spectrometry-based proteomic studies have characterized the glycoproteomic signature of CHL. Furthermore, the sensitivity of ICAM-1 expression in the identification of Reed-Sternberg cells is superior to that of CD15 and maybe useful in the diagnostic workup of CHL. The results of our study have potential applications for the identification of proteins with novel diagnostic and therapeutic relevance.