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Biology subjects

Twells, N.

Publications and source records attributed to Twells, N..

3 recordsLinked to original sources

Age-related remodeling of the glycocalyx drives T cell exhaustion

Cell surface glycans, termed the glycocalyx, are essential regulators of cellular signaling and thus cellular development and functions, but how aging impacts the glycocalyx remains poorly understood. Here, using immune cells as a model system for studying the relationship between aging and glycocalyx remodeling, we show that 2,6-linked sialic acid - a terminal glycan epitope typically associated with inhibitory signaling - becomes downregulated in T cells from older animals. This downregulation is tightly correlated with age-associated accumulation of effector T cells, which are decorated with little to no 2,6-linked sialic acids. T cell aging renders older individuals more vulnerable to infections and cancers. To understand the role of 2,6-linked sialic acids in T cell physiology, we generated a mouse model with T cell-specific deletion of the sialyltransferase gene St6gal1. The chronic depletion of 2,6-linked sialic acids led to naive T (TN) cells expansion in the periphery and premature T cell exhaustion. As a result, these mice were less able to control acute Listeria infection and chronic tumor growth. Blockade of the PD-1 pathway can partially restore the ability of St6gal1-deficient T cells to control tumor growth. Together, these data suggest that 2,6-linked sialic acids are critical for maintaining long-term T cell responsiveness, and the loss of 2,6-linked sialic acids may directly contribute to age-related T cell exhaustion.

immunology↗

Combinatorial CRISPR screens and lectin microarrays identify novel glycosylation regulators

Glycans play critical roles in cellular signaling and function. Unlike proteins, glycan structures are not templated from genes but the concerted activity of many genes, making them historically challenging to study. Here, we present a strategy that utilizes pooled CRISPR screens and lectin microarrays to uncover and characterize regulators of cell surface glycosylation. We applied this approach to study the regulation of high mannose glycans - the starting structure of all asparagine(N)-linked-glycans. We used CRISPR screens to uncover the expanded network of genes controlling high mannose surface levels, followed by lectin microarrays to fully measure the complex effect of select regulators on glycosylation globally. Through this, we elucidated how two novel high mannose regulators - TM9SF3 and the CCC complex - control complex N-glycosylation via regulating Golgi morphology and function. Notably, this method allowed us to interrogate Golgi function in-depth and reveal that similar disruption to Golgi morphology can lead to drastically different glycosylation outcomes. Collectively, this work demonstrates a generalizable approach for systematically dissecting the regulatory network underlying glycosylation.

cell biology↗

The microenvironment dictates glycocalyx construction and immune surveillance

Efforts to identify anti-cancer therapeutics and understand tumor-immune interactions are built with in vitro models that do not match the microenvironmental characteristics of human tissues. Using in vitro models which mimic the physical properties of healthy or cancerous tissues and a physiologically relevant culture medium, we demonstrate that the chemical and physical properties of the microenvironment regulate the composition and topology of the glycocalyx. Remarkably, we find that cancer and age-related changes in the physical properties of the microenvironment are sufficient to adjust immune surveillance via the topology of the glycocalyx, a previously unknown phenomenon observable only with a physiologically relevant culture medium. Key PointsO_LICulture medium dictates cellular mechanoresponse signatures in vitro C_LIO_LIEpithelial glycocalyx construction is mediated by Heat Shock Factor 1 (HSF1) C_LIO_LISialic acid topology dictates Natural Killer cell cytotoxicity C_LIO_LIPhysiological microenvironments reveal distinct glycobiology C_LI

cell biology↗