N‐glycome inheritance from cells to extracellular vesicles in B16 melanomas

Harada, Yoichiro; Kizuka, Yasuhiko; Tokoro, Yuko; Kondo, Kosuke; Yagi, Hirokazu; Kato, Koichi; Inoue, Hiromasa; Taniguchi, Naoyuki; Maruyama, Ikuro

doi:10.1002/1873-3468.13377

Cited by 15 publications

(13 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, we suggest that, in the case of our liposomal exosomes, CTB binds only to GM1; however, in the case of natural membranes (plasma membrane and exosomal membrane), CTB binds to both GM1 and fucosylated glycoproteins, hence the affinity of CTB to natural membranes is higher in comparison with model lipid membranes (lipid bilayers and liposomes). Fucosylated N-glycans were found on glycoproteins in tumor-derived EVs [26][27][28].…”

Section: Discussionmentioning

confidence: 99%

Cholera Toxin Subunit B for Sensitive and Rapid Determination of Exosomes by Gel Filtration

et al. 2020

View full text Add to dashboard Cite

We developed a sensitive fluorescence-based assay for determination of exosome concentration. In our assay, Cholera toxin subunit B (CTB) conjugated to a fluorescence probe and a gel filtration technique (size-exclusion chromatography) are used. Exosomal membranes are particularly enriched in raft-forming lipids (cholesterol, sphingolipids, and saturated phospholipids) and in GM1 ganglioside. CTB binds specifically and with high affinity to exosomal GM1 ganglioside residing in rafts only, and it has long been the probe of choice for membrane rafts. The CTB-gel filtration assay allows for detection of as little as 3 × 108 isolated exosomes/mL in a standard fluorometer, which has a sensitivity comparable to other methods using advanced instrumentation. The linear quantitation range for CTB-gel filtration assay extends over one order of magnitude in exosome concentration. Using 80 nM fluorescence-labeled CTB, we quantitated 3 × 108 to 6 × 109 exosomes/mL. The assay ranges exhibited linear fluorescence increases versus exosome concentration (r2 = 0.987). The assay was verified for exosomal liposomes. The assay is easy to use, rapid, and does not require any expensive or sophisticated instrumentation.

show abstract

Section: Discussionmentioning

confidence: 99%

Cholera Toxin Subunit B for Sensitive and Rapid Determination of Exosomes by Gel Filtration

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Furthermore, the skin cutaneous melanoma has shown the upregulation of the HBP pathway activity as well as single HBP components in comparison with the normal skin (Jia et al 2020 ). On the other hand, the most important enzymes responsible for the N -glycan branching are GnT-III (encoded by gene MGAT3 ), GnT-IV (encoded be two genes MGAT4a and MGAT4b ), GnT-V (encoded by gene MGAT5 ), GnT-IX (encoded by gene MGAT9 ), Fut8 (encoded by gene FUT8 ) and GCNT2 (Taniguchi and Kizuka 2015 ; Harada et al 2019 ; Dimitroff 2019 ). The overexpression of MGAT3 and MGAT5 genes were identified in metastatic melanoma cell line WM266-4 isolated from the lymph node (Bubka et al 2014 ; Link-Lenczowski et al 2018 ).…”

Section: Methodology Used For Investigating Changes In Melanoma Signaling Pathwaysmentioning

confidence: 99%

“…With increasing metastatic potential, cells mostly lacked the I-branched N-glycan antennae which was due to the depressed GCNT2 expression (Sweeney et al 2018 ). In general, the results obtained for melanoma cells demonstrate that the core structure profile of N-glycans may be inherited at the genetic level during the multiple selection cycles of B16 variants (B16–F1, B16–F10, B16–BL6) (Harada et al 2019 ).…”

Section: Methodology Used For Investigating Changes In Melanoma Signaling Pathwaysmentioning

confidence: 99%

See 1 more Smart Citation

Biophysical characterization of melanoma cell phenotype markers during metastatic progression

et al. 2021

View full text Add to dashboard Cite

Melanoma is the most fatal form of skin cancer, with increasing prevalence worldwide. The most common melanoma genetic driver is mutation of the proto-oncogene serine/threonine kinase BRAF; thus, the inhibition of its MAP kinase pathway by specific inhibitors is a commonly applied therapy. However, many patients are resistant, or develop resistance to this type of monotherapy, and therefore combined therapies which target other signaling pathways through various molecular mechanisms are required. A possible strategy may involve targeting cellular energy metabolism, which has been recognized as crucial for cancer development and progression and which connects through glycolysis to cell surface glycan biosynthetic pathways. Protein glycosylation is a hallmark of more than 50% of the human proteome and it has been recognized that altered glycosylation occurs during the metastatic progression of melanoma cells which, in turn facilitates their migration. This review provides a description of recent advances in the search for factors able to remodel cell metabolism between glycolysis and oxidative phosphorylation, and of changes in specific markers and in the biophysical properties of cells during melanoma development from a nevus to metastasis. This development is accompanied by changes in the expression of surface glycans, with corresponding changes in ligand-receptor affinity, giving rise to structural features and viscoelastic parameters particularly well suited to study by label-free biophysical methods.

show abstract

“…Different high-resolution quantitative mass spectrometry methods previously established to elucidate the N -glycome and O -glycome of cell lines have been adapted to the study of EV glycosylation. For example, the N -glycome of EVs isolated from glioma [ 71 ], melanoma [ 84 ] and ovarian carcinoma cell lines [ 72 ] has already been characterized. Additionally, the N -glycome of urinary EVs from prostate cancer patients has also been assessed using other high-resolution glycomic methods [ 85 ].…”

Section: Shortcomings Of Glycomic Approaches and Their Applicationmentioning

confidence: 99%

Glycosylation of Cancer Extracellular Vesicles: Capture Strategies, Functional Roles and Potential Clinical Applications

Martins

Ramos

Freitas

et al. 2021

Cells

View full text Add to dashboard Cite

Glycans are major constituents of extracellular vesicles (EVs). Alterations in the glycosylation pathway are a common feature of cancer cells, which gives rise to de novo or increased synthesis of particular glycans. Therefore, glycans and glycoproteins have been widely used in the clinic as both stratification and prognosis cancer biomarkers. Interestingly, several of the known tumor-associated glycans have already been identified in cancer EVs, highlighting EV glycosylation as a potential source of circulating cancer biomarkers. These particles are crucial vehicles of cell–cell communication, being able to transfer molecular information and to modulate the recipient cell behavior. The presence of particular glycoconjugates has been described to be important for EV protein sorting, uptake and organ-tropism. Furthermore, specific EV glycans or glycoproteins have been described to be able to distinguish tumor EVs from benign EVs. In this review, the application of EV glycosylation in the development of novel EV detection and capture methodologies is discussed. In addition, we highlight the potential of EV glycosylation in the clinical setting for both cancer biomarker discovery and EV therapeutic delivery strategies.

show abstract

N‐glycome inheritance from cells to extracellular vesicles in B16 melanomas

Cited by 15 publications

References 31 publications

Cholera Toxin Subunit B for Sensitive and Rapid Determination of Exosomes by Gel Filtration

Cholera Toxin Subunit B for Sensitive and Rapid Determination of Exosomes by Gel Filtration

Biophysical characterization of melanoma cell phenotype markers during metastatic progression

Glycosylation of Cancer Extracellular Vesicles: Capture Strategies, Functional Roles and Potential Clinical Applications

Contact Info

Product

Resources

About