Clinical importance of high-mannose, fucosylated, and complex N-glycans in breast cancer metastasis

Abstract

BACKGROUND. Although aberrant glycosylation is recognized as a hallmark of cancer, glycosylation in clinical breast cancer (BC) metastasis has not yet been studied. While preclinical studies show that the glycocalyx coating of cancer cells is involved in adhesion, migration, and metastasis, glycosylation changes from primary tumor (PT) to various metastatic sites remain unknown in patients. METHODS. We investigated N-glycosylation profiles in 17 metastatic BC patients from our rapid autopsy program. Primary breast tumor, lymph node metastases, multiple systemic metastases, and various normal tissue cores from each patient were arranged on unique single-patient tissue microarrays (TMAs). We performed mass spectrometry imaging (MSI) combined with extensive pathology annotation of these TMAs, and this process enabled spatially differentiated cell-based analysis of N-glycosylation patterns in metastatic BC. RESULTS. N-glycan abundance increased during metastatic progression independently of BC subtype and treatment regimen, with high-mannose glycans most frequently elevated in BC metastases, followed by fucosylated and complex glycans. Bone metastasis, however, displayed increased core-fucosylation and decreased high-mannose glycans. Consistently, N-glycosylated proteins and N-glycan biosynthesis genes were differentially expressed during metastatic BC progression, with reduced expression of mannose-trimming enzymes and with elevated EpCAM, N-glycan branching, and sialyation enzymes in BC metastases versus PT. CONCLUSION. We show in patients that N-glycosylation of breast cancer cells undergoing metastasis occurs in a metastatic site–specific manner, supporting the clinical importance of high-mannose, fucosylated, and complex N-glycans as future diagnostic markers and therapeutic targets in metastatic BC. FUNDING. NIH grants R01CA213428, R01CA213492, R01CA264901, T32CA193145, Dutch Province Limburg “LINK”, European Union ERA-NET TRANSCAN2-643638.

Metabolic Adaptations in an Endocrine-Related Breast Cancer Mouse Model Unveil Potential Markers of Tumor Response to Hormonal Therapy

“…N -linked and O -linked glycosylation use UDP-GlcNAc as substrate to modify the activity of metabolic enzymes, transcription factors, and several signalling molecules ( 63 ). Furthermore, N -glycosylation has been proven to be elevated in elevated in breast cancer primary tumors, as well as in lymph node metastases and distant metastases compared to healthy mammary tissue ( 64 ).…”

Metabolic Links to Socioeconomic Stresses Uniquely Affecting Ancestry in Normal Breast Tissue at Risk for Breast Cancer

“…Differences in baseline glycosylation in normal breast tissue may be contributing to specific differences in breast stromal biology disproportionally affecting BW. While previous studies have shown that increased fucosylation is associated with malignancy ( 38 , 41 , 88 – 90 ), one study found that a specific fucosylated glycan structure had been shown to be relatively higher in normal breast compared to breast tumors, both from breast cancer patients ( 91 ). Based on the previous findings and our current study, it may not be appropriate to generalize fucosylation patterns in breast tissue as distinct fucosylated structures may have distinct associations in normal versus malignant tissue.…”

“…Fucosylation is a well-known glycosylation modification that has been well characterized in cancers ( 32 , 35 , 38 , 48 , 49 , 92 , 93 ). Variations in fucosylation linkages such as outer-arm fucosylation and core-fucosylation have distinct implications for cancer biology ( 27 , 28 , 32 , 35 , 38 , 42 , 92 ). In fact, increased fucosylation, both core and branched segments, have been observed in malignant transformation in many cancers such as breast ( 38 , 41 ), liver ( 66 ), pancreatic ( 94 ), prostate ( 61 ) and colorectal cancers ( 95 ).…”

“…Over many cancer types, changes to N-glycan sugar residue composition and branching structure contributes to processes including stroma-cell adhesive interactions, migration, immune recruitment, and malignant conversion ( 27 , 29 , 31 , 33 , 48 ). Increases in branching, outer arm fucosylation, and sialylation have all been associated with clinical cases of breast cancer and link to breast cancer subtypes ( 40 – 42 , 49 ). In breast cancer, modifications of cell surface 2,6-sialylation alters cell-cell and cell-extracellular matrix (ECM) adhesion ( 50 ); loss of adhesive interactions is one of the first steps in metastatic pathways ( 31 , 51 ).…”

Nomograms Based on Serum N-glycome for Diagnosis of Papillary Thyroid Microcarcinoma and Prediction of Lymph Node Metastasis

“…Our investigation of the dysregulation of N -glycan patterns in PTMC may point to pathophysiological processes involving multiple proteins, as we discuss below. Early studies showed that the high-mannose glycans and complex-type glycans were increased in breast cancer progression, and glycans took part in cancer progression phases, such as cell proliferation signaling, activating invasion and metastasis, and tumor-promoting inflammation [ 36 , 37 , 38 ]. Chik et al have shown high-mannose structures as the most abundant glycan in colorectal cancer cell lines and tissues [ 39 ].…”