Differentiation of Cancer Cell Origin and Molecular Subtype by Plasma Membrane N-Glycan Profiling

Abstract

In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. Glycomic profiling of the cell membrane offers an alternate route towards cancer diagnosis. In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. Based simply on the relative abundances of broad glycan classes (e.g. high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.) most cell lines were readily differentiated. More closely-related cell lines were differentiated based on several-fold differences in the abundances of individual glycans. Based on characteristic N-glycan profiles, primary cancer origins and molecular subtypes could be distinguished. These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications towards cancer diagnosis and direction of treatment.

Human protein paucimannosylation: cues from the eukaryotic kingdoms

“…Recent glycomics and glycoproteomics‐centric studies have demonstrated that PMPs are also elevated glycosylation signatures associated with various human cancers including those of ovarian (Everest‐Dass et al ., ; Chen et al ., ), colorectal (Joosten et al ., ; Balog et al ., ; Sethi et al ., ; Kaprio et al ., ; Holst et al ., ), breast (Lee et al ., ; Chen et al ., ), lung (Hua et al ., ; Ruhaak et al ., ; Wang et al ., ), paraganglioma (Leijon et al ., ), glioblastoma multiforme (Becker et al ., ), skin (Moginger et al ., ) and prostate (Shah et al ., ) origin. By systematically interrogating a very large collection of nearly 500 N ‐glycomics LC‐MS/MS data sets obtained from 11 cancer types and subtypes, we have obtained solid evidence that PMGs are significant features of human cancers and, at least in part, expressed on cancer cell surfaces (S. Chatterjee, L. Y. Lee, R. Kawahara, J. L. Abrahams, B. Adamczyk, M. Anugraham, C. Ashwood, Z. Sumer‐Bayraktar, M. T. Briggs, J. H. L. Chik, A. Everest‐Dass, S. Förster, H. Hinneburg, K. R. M. Leite, I. Loke, U. Möginger, E. S. X. Moh, M. Nakano, S. Recuero, M. K. Sethi, M. Srougi, K. Stavenhagen, V. Venkatakrishnan, K. Wongtrakul‐Kish, S. Diestel, P. Hoffmann, N. G. Karlsson, D. Kolarich, M. P. Molloy, M. H. Muders, M. K. Oehler, N. H. Packer, G. Palmisano & M. Thaysen‐Andersen, in preparation).…”

Broad and thematic remodeling of the surface glycoproteome on isogenic cells transformed with driving proliferative oncogenes

“…There is abundant evidence to suggest that high-mannose type glycans are present at the cell surface, and furthermore, that cancerous cells display increased abundance of highmannose glycans at their cell surface (Holst et al, 2016;Hua et al, 2014). A similar observation was reported in the glycomic comparison of transformed versus human embryonic stem cells (hESC), where high-mannose glycans were observed at significantly higher abundance on plasma membranes of hESCs .…”

Integrated Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts

“…High mannose N-glycans could be an indicator of an embryonic, undifferentiated phenotype 45 . Increased expression of high mannose type N-glycans also have been observed in colorectal cancer cell lines of varying malignancy 46-48 and in breast cancer cell lines and tissue 49-51 . The biological significance of this glycomic alteration in cancer is not clear, yet the presence of high (truncated) mannose indicates some level of incomplete N-linked glycosylation prossessing 52 .…”