Journal
DEVELOPMENTAL CELL
Volume 50, Issue 1, Pages 57-+Publisher
CELL PRESS
DOI: 10.1016/j.devcel.2019.04.035
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Funding
- National Institute of General Medical Sciences [R35GM118067]
- National Institute of Biomedical Imaging and Bioengineering [U01EB021237]
- US National Cancer Institute [R01CA227942]
- US National Institute of General Medical Sciences [R37GM058867]
- National Science Foundation Graduate Research Fellowship
- Stanford Graduate Fellowship
- Stanford ChEM-H Chemistry/Biology Interface Predoctoral Training Program
- Mary Lake Polan Endowment
- Swedish Research Council [2016-00130]
- Foundation BLANCE-FLOR Boncompagni-Ludovisi, nee Bildt
- Mary Lake Polan Gynecologic Oncology Endowment for Gynecologic Cancer Research, Stanford School of Medicine, Stanford University
- Swedish Research Council [2016-00130] Funding Source: Swedish Research Council
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The mammalian glycocalyx is a heavily glycosylated extramembrane compartment found on nearly every cell. Despite its relevance in both health and disease, studies of the glycocalyx remain hampered by a paucity of methods to spatially classify its components. We combine metabolic labeling, bioorthogonal chemistry, and super-resolution localization microscopy to image two constituents of cell-surface glycans, N-acetylgalactosamine (GalNAc) and sialic acid, with 10-20 nm precision in 2D and 3D. This approach enables two measurements: glycocalyx height and the distribution of individual sugars distal from the membrane. These measurements show that the glycocalyx exhibits nanoscale organization on both cell lines and primary human tumor cells. Additionally, we observe enhanced glycocalyx height in response to epithelial-to-mesenchymal transition and to oncogenic KRAS activation. In the latter case, we trace increased height to an effector gene, GALNT7. These data highlight the power of advanced imaging methods to provide molecular and functional insights into glycocalyx biology.
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