The Role of Glycan Structures in Modulating GM-CSF Bioactivity: Insights from Glycoengineering

Cagdas, E.; Skovbakke, S. L.; Agullet, J. P.; Dworkin, L. A.; Scapin, G.; Hefzi, H.; Fremming, K. S.; Schoffhelen, S.; Putkaradze, N.; Voldborg, B.; Grav, L. M.; Nielsen, L. K.; Goletz, S. G.; Lewis, N. E.

doi:10.64898/2026.02.17.706267

bioRxiv · 10.64898/2026.02.17.706267

The Role of Glycan Structures in Modulating GM-CSF Bioactivity: Insights from Glycoengineering

Cagdas, E.·Skovbakke, S. L.·Agullet, J. P.·Dworkin, L. A.·Scapin, G.·Hefzi, H.·Fremming, K. S.·Schoffhelen, S.·Putkaradze, N.·Voldborg, B.·Grav, L. M.·Nielsen, L. K.·Goletz, S. G.·Lewis, N. E.

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Abstract

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a glycoprotein cytokine with therapeutic potential in cancer and neutropenia treatment. While glycosylation of GM-CSF reduces immunogenicity and enhances serum bioavailability, it can also diminish receptor binding and bioactivity. Based on transcriptomic analysis of human T lymphocytes reported previously, GM-CSF-producing cells exhibit elevated expression of Alpha-1,6-Mannosylglycoprotein 6-Beta-N-Acetylglucosaminyltransferase (MGAT5), which encodes N-acetylglucosaminyltransferase V, an enzyme involved in N-glycan branching. Given this role of MGAT5 in glycosylation, we produced GM-CSF variants using glycoengineered Chinese hamster ovary cells to generate diverse glycoforms and assessed their bioactivity. Testing their activity on TF-1 cell proliferation, we found that decreases in GM-CSF N-glycan branching significantly suppressed its activity. These findings underscore the importance of glycosylation in modulating the efficacy and safety of GM-CSF-based therapeutics, suggesting that precise glycoengineering may be key to optimizing GM-CSF performance in clinical applications.

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Cagdas, E., Skovbakke, S. L., Agullet, J. P., Dworkin, L. A., Scapin, G., Hefzi, H., Fremming, K. S., Schoffhelen, S., Putkaradze, N., Voldborg, B., Grav, L. M., Nielsen, L. K., Goletz, S. G., Lewis, N. E.. 2026-02-18. The Role of Glycan Structures in Modulating GM-CSF Bioactivity: Insights from Glycoengineering. https://doi.org/10.64898/2026.02.17.706267

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