In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation

Manveen K. Sethi, Hoguen Kim, Cheol Keun Park, Mark S. Baker, Young-Ki Paik, Nicolle H. Packer, William S. Hancock, Susan Fanayan, Morten Thaysen-Andersen

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. Interestingly, a novel link between the EGFR status and the N-glycosylation was identified using hierarchical clustering of the N-glycome profiles. EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC.

Original languageEnglish
Pages (from-to)1064-1078
Number of pages15
JournalGlycobiology
Volume25
Issue number10
DOIs
Publication statusPublished - 2015 May 4

Fingerprint

Glycosylation
Epidermal Growth Factor Receptor
Polysaccharides
Colorectal Neoplasms
Tissue
Neoplasms
Proteins
Glycomics
Deregulation
Liquid chromatography
Proteome
Mannose
Mass spectrometry
Labels
Tumors
Glycoproteins
Carbon
Membranes
Tandem Mass Spectrometry
Liquid Chromatography

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Sethi, Manveen K. ; Kim, Hoguen ; Park, Cheol Keun ; Baker, Mark S. ; Paik, Young-Ki ; Packer, Nicolle H. ; Hancock, William S. ; Fanayan, Susan ; Thaysen-Andersen, Morten. / In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. In: Glycobiology. 2015 ; Vol. 25, No. 10. pp. 1064-1078.
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abstract = "Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. Interestingly, a novel link between the EGFR status and the N-glycosylation was identified using hierarchical clustering of the N-glycome profiles. EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC.",
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Sethi, MK, Kim, H, Park, CK, Baker, MS, Paik, Y-K, Packer, NH, Hancock, WS, Fanayan, S & Thaysen-Andersen, M 2015, 'In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation', Glycobiology, vol. 25, no. 10, pp. 1064-1078. https://doi.org/10.1093/glycob/cwv042

In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. / Sethi, Manveen K.; Kim, Hoguen; Park, Cheol Keun; Baker, Mark S.; Paik, Young-Ki; Packer, Nicolle H.; Hancock, William S.; Fanayan, Susan; Thaysen-Andersen, Morten.

In: Glycobiology, Vol. 25, No. 10, 04.05.2015, p. 1064-1078.

Research output: Contribution to journalArticle

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T1 - In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation

AU - Sethi, Manveen K.

AU - Kim, Hoguen

AU - Park, Cheol Keun

AU - Baker, Mark S.

AU - Paik, Young-Ki

AU - Packer, Nicolle H.

AU - Hancock, William S.

AU - Fanayan, Susan

AU - Thaysen-Andersen, Morten

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