Development of a glycan-based platform for the elucidation cell-glycosaminoglycans interactions

last updated: 2018-06-08
ProjectComplexiTE :: publications list
TitleDevelopment of a glycan-based platform for the elucidation cell-glycosaminoglycans interactions
Publication TypeComunications - Poster
Year of Publication2017
AuthorsCarvalho A. M., Soares da Costa D., Reis R. L., and Pashkuleva I.
Abstract

The chemical and structural diversity, molecular weight (MW) and patterns of glycosaminoglycans (GAGs) in the extracellular matrix (ECM) are important factors in cells' stemness, differentiation, development, and thus, in the advancement and severity of different diseases [1, 2]. In the ECM, GAGs are commonly presented as conjugates with proteins or lipids. These conjugates are prone to interact with transmembrane proteins and can have different bioroles, including physiological barriers, storage depots of soluble factors, and modulators cell-ECM and cell-cell interactions. The binding of GAGs to their receptors is driven by weak and often multivalent interactions resulting in receptors clustering. Generally, these interactions are hard to detect and measure [3, 4]. State of the art studies involved the use of GAGs in solution – state that is associated with higher freedom of mobility as compared with the native GAGs immobilized in ECM.

Herein, we propose an alternative design that is based on end-on immobilized GAGs. The developed platform is versatile as it allows immobilization of different GAGs and offers a possibility to study their interactions with cells in a high-throughput fashion using complementary label-free techniques. We used APTES silanized glass as a substrate for the platform development. Continuous colloidal gradients were assembled on this substrate using gold nanoparticles (20 nm). GAGs, functionalized with alkanthiol (C11SH) at the reducing end were covalently bound to the gold gradients as confirmed by water contact angle, x-ray spectroscopy (XPS), and fluorescence microscopy. The stability and biofunctionality of the immobilized GAGs was confirmed by studying their interactions with lectin - Wheat germ agglutinin that is specific towards N-acetyl-D-glucosamine was used in these studies. Preliminary evaluation of the interactions of hyaluronan platforms with 3 breast cancer cell lines that differ by their expression of CD44 were also performed. The tested cells recognize the gradient and more attached cells were observed at higher densities of hyaluronan. Cell morphology was also affected - cells typically spread on areas richer in hyaluronic acid. Of note, these differences were more pronounced for cells with lower expression of CD44, thus, allowing further application of the developed platforms for cell separation.

References:

1.            Costa, D.S., R.L.R. Reis, and I. Pashkuleva, Sulfation of Glycosaminoglycans and Its Implications in Human Health and Disorders. Annual Review of Biomedical Engineering, 2017. 19(1): p. 1-26.

2.            Bülow, H.E. and O. Hobert, The Molecular Diversity of Glycosaminoglycans Shapes Animal Development. Annual Review of Cell and Developmental Biology, 2006. 22(1): p. 375-407.

3.            Crocker, P.R. and T. Feizi, Carbohydrate recognition systems: functional triads in cell—cell interactions. Current Opinion in Structural Biology, 1996. 6(5): p. 679-691.

4.            Paulson, J.C., O. Blixt, and B.E. Collins, Sweet spots in functional glycomics. Nat Chem Biol, 2006. 2(5): p. 238-248.

Conference NameFoReCaST - The Tumour Microenvironment
Date Published2017-11-15
Conference LocationPorto, Portugal
KeywordsCD44, high-throughput, Hyaluronic acid
RightsopenAccess
Peer reviewedno
Statuspublished

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