Genetic and small molecule strategies to edit the glycocalyx

Abstract

The glycocalyx covers all living cells and is a key regulator of cell communication with the extracellular milieu/matrix, other cells, foreign pathogens and the microbiome. A variety of glycoconjugate types decorate mammalian cells including N‐ and O‐linked glycans attached to proteins, glycolipids, and glycosaminoglycans. A complete understanding of the molecular and cellular features controlling living systems is not possible without delineating the structural and functional contributions of the glycocalyx. My laboratory studies this aspect by developing systems based experimental and computational tools to characterize the role of the glycocalyx in normal physiological and selected pathophysiological contexts. For such investigations, we have developed both novel CRISPR‐Cas9 based tools to perturb cell surface glycans and small molecule inhibitors of the same. Additionally, we have developed a Synthetic Glycan toolkit and liquid‐chromatography mass‐spectrometry (LC‐MS) workflow to longitudinally track glycan changes in living cells in a non‐destructive manner. My talk will present the development and application of these technologies in two contexts. First, I will discuss studies of human inflammatory diseases which will present some new molecular players regulating glycosylation and leukocyte‐endothelial cell adhesion mechanics under fluid shear flow. Second, I will present our studies on the role of glycans in regulating SARS‐CoV‐2 viral entry into human cells expressing ACE2. The data demonstrate a critical role for N‐glycans and O‐linked glycans in viral entry and propagation.