Detecting the Sweet Biomarker on Cancer Cells - ACS Central Science

Mar 29, 2018 - ... and Peking-Tsinghua Center for Life Sciences, Peking University, ... Copyright © 2018 American Chemical Society ... ACS Central Sc...
2 downloads 0 Views 694KB Size
This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.

Reactions FIRSTFirst REACTIONS

Detecting the Sweet Biomarker on Cancer Cells Xing Chen College of Chemistry and Molecular Engineering, and Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, 100871, China

called glycosyltransferases that can specifically recognize a glycan acceptor and transfer on it a monosaccharide analogue via its corresponding nucleotide sugar donor (Figure 2C). When the monosaccharide analogue contains a bioorthogonal group, a second-step click reaction can again be performed to install various probes. Several glycan epitopes have been probed using CeGL, including LacNAc, Fucα2Gal, and TF antigen.6 In this issue of ACS Central Science,1 Wen and co-workers used CeGL to detect sialyl-T by searching for a glycosyltransferase that only recognizes sialyl-T as the acceptor, but with relaxed standards for the donor. They found a human α-N-acetylgalactosaminide sialyltransferase (ST6GalNAc IV) that has high specificity for sialyl-T and is able to transfer biotin-functionalized sialic acid analogues (Figure 1B). The use of biotin, rather than a chemical handle, makes this a one-step labeling process, which is an appealing feature of ST6GalNAc. The key is ST6GalNAc’s broad tolerance for large modifications on sugar donors. Mbua et al. earlier reported that another sialyltransferase, ST6Gal-I, can also transfer sialic acid analogues bearing large functional groups and be used for one-step CeGL labeling of N-linked glycans.7 Wen and coauthors suggest that the tolerance of sugar donors with large modification groups might be a common feature shared by sialyltransferases.1

A sugar molecule expressed on cancer cells can be chemoenzymatically labeled for fluorescent imaging and proteomic analysis.

T

he ability to detect cancer biomarkers (i.e., characteristic biomolecules appearing on cancer cells but not on normal cells) is crucial for early diagnosis of cancers. Now, a research team led by Liuqing Wen and Peng George Wang at Georgia State University provides a chemical tool for labeling and visualizing an antigen that is highly expressed on the surfaces of various cancer cells.1 Cells are coated with a dense layer of sugars (also called carbohydrates or glycans), and the cancer cells are no exception. Quite often, cancer cells carry specific glycan epitopes that either are not expressed, or only minimally so, in normal tissues. One of these cancer-specific glycans is sialylated Thomsen-Freidenreich, or sialyl-T, a trisaccharide found on mucin-type O-linked glycoproteins (Figure 1A). Accumulating evidence indicates that the presence of sialyl-T is correlated strongly with tumor development and progression.2 Unfortunately, detecting sialyl-T on live cells has been difficult until now due to the lack of an effective traditional testing method, such as an antibody or lectin (Figure 2A). In addition, antibodies and lectins often suffer from relatively low specificity and affinity. An alternative approach involves using cellular metabolism to label glycans with a monosaccharide analogue containing a bioorthogonal functional group such as an azide or alkyne, which can be subsequently conjugated with imaging probes or affinity tags using click chemistry (Figure 2B). This two-step chemical method has become a primary tool in the detection of glycans on live cells and in living organisms.3,4 However, this sort of metabolic labeling can only target monosaccharides, such as sialic acid. Although sialyl-T contains a sialic acid, other glycans, including those found on the surface of healthy cells, also bear this monosaccharide. A recently emerged technique, chemoenzymatic glycan labeling (CeGL), addresses this limitation.5,6 The CeGL strategy exploits a class of enzymes © XXXX American Chemical Society

The use of biotin, rather than a chemical handle, makes this a one-step labeling process, which is an appealing feature of ST6GalNAc. The key is ST6GalNAc’s broad tolerance for large modifications on sugar donors. By using the ST6GalNAc IV-based CeGL, the authors visualized the expression of sialyl-T on a variety of cancer cells by fluorescence microscopy and quantitatively compared the expression level by flow cytometry. Furthermore, by using mass

A

DOI: 10.1021/acscentsci.8b00156 ACS Cent. Sci. XXXX, XXX, XXX−XXX

Reactions FIRSTFirst REACTIONS

ACS Central Science

Figure 1. One-step CeGL labeling of sialyl-T. (A) Chemical structure of sialyl-T α-linked to the serine or threonine residue of cell-surface proteins (purple diamond−yellow circle−yellow square). (B) Living cells were treated with ST6GalNAc-IV and a CMP-sialic acid analogue containing a biotin moiety. ST6GalNAc-IV specifically recognizes sialyl-T on the cell surface and transfers biotin-functionalized sialic acid onto the GalNAc (yellow circle) of sialyl-T. The biotin can then be stained with streptavidin-fluorophore for imaging or streptavidin beads for enrichment and glycoproteomic profiling.

Figure 2. Three methods for labeling cell-surface glycans. (A) Antibodies and lectins conjugates with fluorescent probes can be used to directly detect glycans on cell surfaces. (B) MGL exploits the cellular machinery to metabolically incorporate monosaccharide analogues containing a bioorthogonal functional group, X, into cell-surface glycans. In the second step, a probe containing a complementary bioorthogonal group, Y, is reacted with X to label the glycans. (C) CeGL uses glycosyltransferases that have stringent acceptor specificity but can transfer monosaccharide analogues from the corresponding nucleotide sugars.

glycotransferases have recently been successfully injected into living mice to enzymatically modulate glycosylation of endogenous antibodies.8

spectrometry-based proteomic analysis, they identified 78 cellsurface proteins in human breast cancer cells and 43 proteins in human colon cancer cells that are modified with sialyl-T.1 These results provide valuable information for understanding the biological function of sialyl-T in cancer progression. Further evaluation is needed to determine whether CeGL can be used in living animals. Encouragingly, engineered

Author Information

E-mail: [email protected]. ORCID

Xing 0000-0002-3058-7370 XingChen: Chen : 0000-0002-3058-7370 B

DOI: 10.1021/acscentsci.8b00156 ACS Cent. Sci. XXXX, XXX, XXX−XXX

Reactions FIRSTFirst REACTIONS

ACS Central Science Notes

The author declares no competing financial interest.



REFERENCES REFERENCES (1) Wen, L.; Liu, D.; Zheng, Y.; Huang, K.; Cao, X.; Song, J.; Wang, P. G. A One-Step Chemoenzymatic Labeling Strategy for Probing Sialylated Thomsen−Friedenreich Antigen. ACS Cent. Sci. 2018, DOI: 10.1021/acscentsci.7b00573. (2) Remmers, N.; Anderson, J. M.; Linde, E. M.; DiMaio, D. J.; Lazenby, A. J.; Wandall, H. H.; Mandel, U.; Clausen, H.; Yu, F.; Hollingsworth, M. A. Aberrant expression of mucin core proteins and o-linked glycans associated with progression of pancreatic cancer. Clin. Cancer Res. 2013, 19, 1981−1993. (3) Laughlin, S. T.; Bertozzi, C. R. Imaging the glycome. Proc. Natl. Acad. Sci. U. S. A. 2009, 106, 12−17. (4) Cheng, B.; Xie, R.; Dong, L.; Chen, X. Metabolic Remodeling of Cell-Surface Sialic Acids: Principles, Applications, and Recent Advances. ChemBioChem 2016, 17, 11−27. (5) Griffin, M. E.; Hsieh-Wilson, L. C. Glycan Engineering for Cell and Developmental Biology. Cell Chemical Biology 2016, 23, 108−121. (6) López-Aguilar, A.; Briard, J. G.; Yang, L.; Ovryn, B.; Macauley, M. S.; Wu, P. Tools for Studying Glycans: Recent Advances in Chemoenzymatic Glycan Labeling. ACS Chem. Biol. 2017, 12, 611− 621. (7) Mbua, N. E.; Li, X.; Flanagan-Steet, H. R.; Meng, L.; Aoki, K.; Moremen, K. W.; Wolfert, M. A.; Steet, R.; Boons, G.-J. Selective exoenzymatic labeling of N-glycans on the surface of living cells by recombinant ST6Gal I. Angew. Chem., Int. Ed. 2013, 52, 13012−13015. (8) Pagan, J. D.; Kitaoka, M.; Anthony, R. M. Engineered Sialylation of Pathogenic Antibodies In Vivo Attenuates Autoimmune Disease. Cell 2018, 172, 564−577.e13 e13.

C

DOI: 10.1021/acscentsci.8b00156 ACS Cent. Sci. XXXX, XXX, XXX−XXX