Sialic Acid as a Biomarker Studied in Breast Cancer Cell Lines In Vitro Using Fluorescent Molecularly Imprinted Polymers

El-Schich, Zahra; Zhang, Yuecheng; Göransson, Tommy; Dizeyi, Nishtman; Persson, Jenny L.; Johansson, E. Susanne; Caraballo, Rémi; Elofsson, Mikael; Shinde, Sudhirkumar; Sellergren, Börje; Wingren, Anette Gjörloff

doi:10.3390/app11073256

Cited by 13 publications

(10 citation statements)

References 37 publications

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“…Our binding results show that SA-MIPs with a silica-coated PS core have a more pronounced tendency to conjugate with cancer cells, presumably due to their smaller size and better suspensibility. Previous work in our group using other SA-MIP batches with a larger size did not show the same binding efficiency [103,[119][120][121]. Optimization of the SA-MIPs synthesis protocol improved the binding features and specificity as shown by our results, and in a related paper by Kimani et al [81].…”

Section: Binding Of Lectins and Sa-mips To Cancer Cell Linessupporting

confidence: 75%

Connecting sialic acid expression to cancer cell characteristics: Novel tools for detection, imaging, and analysis

Sternbæk¹

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Sialic acid (SA) plays a crucial role in many biological processes. Cell surface SA expression is usually analyzed with antibodies or lectins; however, they are costly and with poor stability. We have used a molecular imprinting technique to synthesize an alternative SA receptor – SA molecularly imprinted polymers(SA-MIPs) with an embedded fluorophore for fluorescent detection of theSA-MIPs. The binding behavior and specificity of SA-MIPs were verified by using lectins and SA conjugates on cancer cell lines, showing that SA-MIPs can be used as an effective tool for SA expression analysis of cancer cells. Digital holographic cytometry (DHC) is a non-phototoxic quantitative phase imaging technique that facilitates the monitoring of living cells over time. We have demonstrated the potential of DHC by mapping cellular parameters, such as cell number, area, thickness, and volume. In addition, cellular parameters possibly depending on sialylation, were evaluated using DHC. Furthermore, the uptake over time of SA-MIPs by macrophages was investigated for any inflammatory and/or cytotoxic responses when administered to phagocytosing cells. Our results indicate that SA-MIPs caused low induction and sparse secretion of inflammatory cytokines, and that reduced cell proliferation was not due to cytotoxicity, but to attenuated cell cycles. These results suggest that SA-MIPs will contribute to the further understanding of cancer cell behavior and can be an asset for in vivo studies.

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Section: Binding Of Lectins and Sa-mips To Cancer Cell Linessupporting

confidence: 75%

Connecting sialic acid expression to cancer cell characteristics: Novel tools for detection, imaging, and analysis

Sternbæk¹

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“…This was unexpected as B3GNT2 is reported to act on LacNAc acceptors on Nglycans, O-glycans, and glycolipids. 29 Since many cancer cell lines have truncated O-glycan structures, 55,56 we hypothesized that the apparent N-glycan specificity was attributed to the HS-578-T breast cancer cell line not displaying extended O-glycan structures that could serve as suitable acceptor sites for B3GNT2. Thus, GlcNAz-engineered HEK293T cells were also analyzed by streptavidin immunoblotting (Figure 4D).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Glyco-Engineering Cell Surfaces by Exo-Enzymatic Installation of GlcNAz and LacNAz Motifs

De León González,

Boddington,

Kofsky

et al. 2024

ACS Chem. Biol.

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Exo-enzymatic glyco-engineering of cell-surface glycoconjugates enables the selective display of well-defined glyco-motifs bearing bioorthogonal functional groups, which can be used to study glycans and their interactions with glycan-binding proteins. In recent years, strategies to edit cellular glycans by installing monosaccharides and their derivatives using glycosyltransferase enzymes have rapidly expanded. However, analogous methods to introduce chemical reporter-functionalized type 2 LacNAc motifs have not been reported. Herein, we report the chemo-enzymatic synthesis of unnatural UDP-GlcNAc and UDP-GalNAc nucleotide-sugars bearing azide, alkyne, and diazirine functionalities on the C2-acetamido group using the mutant uridylyltransferase AGX1 F383A . The unnatural UDP-GlcNAc derivatives were examined as substrates for the human GlcNAc-transferase B3GNT2, where it was found that modified donors were tolerated for transfer, albeit to a lesser extent than the natural UDP-GlcNAc substrate. When the GlcNAc derivatives were examined as acceptor substrates for the human Gal-transferase B4GalT1, all derivatives were well tolerated and the enzyme could successfully form derivatized LacNAcs. B3GNT2 was also used to exo-enzymatically install GlcNAc and unnatural GlcNAc derivatives on cell-surface glycans. GlcNAc-or GlcNAz-engineered cells were further extended by B4GalT1 and UDP-Gal, producing LacNAc-or LacNAz-engineered cells. Our proof-of-concept glyco-engineering labeling strategy is amenable to different cell types and our work expands the exo-enzymatic glycan editing toolbox to selectively introduce unnatural type 2 LacNAc motifs.

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“…This finding confirms the selectivity of the MIP particles for binding SA. We recently demonstrated the novel use of SA conjugates ME1057 and ME0970 by pre-treating the SA-MIPs (200 nm) with these SA conjugates as inhibitors [30]. It has also been shown that the titration of SA-MIPs with possible competitors GalNAc and glucose did not induce a fluorescence increase [22].…”

Section: Discussionmentioning

confidence: 99%

Fluorescent Molecularly Imprinted Polymer Layers against Sialic Acid on Silica-Coated Polystyrene Cores—Assessment of the Binding Behavior to Cancer Cells

Beyer

Kimani

Zhang

et al. 2022

Cancers

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Sialic acid (SA) is a monosaccharide usually linked to the terminus of glycan chains on the cell surface. It plays a crucial role in many biological processes, and hypersialylation is a common feature in cancer. Lectins are widely used to analyze the cell surface expression of SA. However, these protein molecules are usually expensive and easily denatured, which calls for the development of alternative glycan-specific receptors and cell imaging technologies. In this study, SA-imprinted fluorescent core-shell molecularly imprinted polymer particles (SA-MIPs) were employed to recognize SA on the cell surface of cancer cell lines. The SA-MIPs improved suspensibility and scattering properties compared with previously used core-shell SA-MIPs. Although SA-imprinting was performed using SA without preference for the α2,3- and α2,6-SA forms, we screened the cancer cell lines analyzed using the lectins Maackia Amurensis Lectin I (MAL I, α2,3-SA) and Sambucus Nigra Lectin (SNA, α2,6-SA). Our results show that the selected cancer cell lines in this study presented a varied binding behavior with the SA-MIPs. The binding pattern of the lectins was also demonstrated. Moreover, two different pentavalent SA conjugates were used to inhibit the binding of the SA-MIPs to breast, skin, and lung cancer cell lines, demonstrating the specificity of the SA-MIPs in both flow cytometry and confocal fluorescence microscopy. We concluded that the synthesized SA-MIPs might be a powerful future tool in the diagnostic analysis of various cancer cells.

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Sialic Acid as a Biomarker Studied in Breast Cancer Cell Lines In Vitro Using Fluorescent Molecularly Imprinted Polymers

Cited by 13 publications

References 37 publications

Connecting sialic acid expression to cancer cell characteristics: Novel tools for detection, imaging, and analysis

Connecting sialic acid expression to cancer cell characteristics: Novel tools for detection, imaging, and analysis

Glyco-Engineering Cell Surfaces by Exo-Enzymatic Installation of GlcNAz and LacNAz Motifs

Fluorescent Molecularly Imprinted Polymer Layers against Sialic Acid on Silica-Coated Polystyrene Cores—Assessment of the Binding Behavior to Cancer Cells

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