A framework for the simulation of individual glycan coordinates to analyze spatial relationships within the...

The glycocalyx is a dense and dynamic layer of glycosylated species that covers every cell in the human body. It plays crucial roles in various cellular processes in health and disease, such as cancer immune evasion, cancer immune therapy, blastocyst implantation, and functional attenuation of membrane protein diffusion. In addition, alterations in glycocalyx structure may play an important role in ocular surface diseases, e.g., dry eye disease. Despite the emerging importance of the glycocalyx, various aspects of its functional organization remain elusive to date. A central reason for this elusiveness is the nanoscale dimension of the glycocalyx in conjunction with its high structural complexity, which is not accessible to observation with conventional light microscopy. Recent advances in super-resolution microscopy have enabled resolutions down to the single-digit nanometer range. In order to fully leverage the potential of these novel methods, computational frameworks that allow for contextualization of the resulting experimental data are required. Here, we present a simulation-based approach to analyze spatial relationships of glycan components on the cell membrane based on known geometrical parameters. We focus on sialic acids in this work, but the technique can be adapted to any glycan component of interest. By integrating data from mass spectrometry and quantitative biological studies, these simulations aim to model possible experimental outcomes, which can then be used for further analysis, such as spatial point statistics. Importantly, we include various experimental considerations, such as labeling and detection efficiency. This approach may contribute to establishing a new standard of connection between geometrical and molecular-resolution data in service of advancing our understanding of the functional role of the glycocalyx in biology as well as its clinical potential.