N-glycosylation by Mgat5 imposes a targetable constraint on immune-mediated tumor clearance
Erin E. Hollander, Rosemary E. Flock, Jayne C. McDevitt, William P. Vostrejs, Sydney L. Campbell, Margo I. Orlen, Samantha B. Kemp, Benjamin M. Kahn, Kathryn E. Wellen, Il-Kyu Kim, Ben Z. Stanger
Erin E. Hollander, Rosemary E. Flock, Jayne C. McDevitt, William P. Vostrejs, Sydney L. Campbell, Margo I. Orlen, Samantha B. Kemp, Benjamin M. Kahn, Kathryn E. Wellen, Il-Kyu Kim, Ben Z. Stanger
View: Text | PDF
Research Article Cell biology Oncology

N-glycosylation by Mgat5 imposes a targetable constraint on immune-mediated tumor clearance

Abstract

The regulated glycosylation of the proteome has widespread effects on biological processes that cancer cells can exploit. Expression of N-acetylglucosaminyltransferase V (encoded by Mgat5 or GnT-V), which catalyzes the addition of β1,6-linked N-acetylglucosamine to form complex N-glycans, has been linked to tumor growth and metastasis across tumor types. Using a panel of murine pancreatic ductal adenocarcinoma (PDAC) clonal cell lines that recapitulate the immune heterogeneity of PDAC, we found that Mgat5 is required for tumor growth in vivo but not in vitro. Loss of Mgat5 results in tumor clearance that is dependent on T cells and dendritic cells, with NK cells playing an early role. Analysis of extrinsic cell death pathways revealed Mgat5-deficient cells have increased sensitivity to cell death mediated by the TNF superfamily, a property that was shared with other non-PDAC Mgat5-deficient cell lines. Finally, Mgat5 knockout in an immunotherapy-resistant PDAC line significantly decreased tumor growth and increased survival upon immune checkpoint blockade. These findings demonstrate a role for N-glycosylation in regulating the sensitivity of cancer cells to T cell killing through classical cell death pathways.

Authors

Erin E. Hollander, Rosemary E. Flock, Jayne C. McDevitt, William P. Vostrejs, Sydney L. Campbell, Margo I. Orlen, Samantha B. Kemp, Benjamin M. Kahn, Kathryn E. Wellen, Il-Kyu Kim, Ben Z. Stanger

×

Figure 3

Mgat5 loss increases the immunogenicity of existing tumor antigens.

Options: View larger image (or click on image) Download as PowerPoint
Mgat5 loss increases the immunogenicity of existing tumor antigens. (A)...
(A) Vβ repertoire analysis of CD4+ and CD8+ T cells in the spleens of naive mice (n = 5, combined from 2 independent experiments), mice bearing 2838c3 WT tumors (n = 3), or mice that had cleared Mgat5-KO-A tumors (n = 5). Data represent mean ± SEM. Statistical analysis by 2-way ANOVA for this and all further tumor growth curves. (B) Growth (mm3) of subcutaneous tumors arising from 2838c3 WT cells injected into either naive mice or mice previously immunized (4 weeks earlier) with 2838c3 Mgat5-KO-A cells (n = 6 mice/group). Data represent mean ± SEM. (C) Growth (mm3, left) and weights (right) of 6694c2 WT cells subcutaneous injected into either naive mice or mice previously immunized (4 weeks earlier) with 2838c3 Mgat5-KO-A cells (n = 6 mice/group). Data represent mean ± SEM. Statistical analysis by unpaired, 2-tailed Student t test for tumor weights. (D) Growth (mm3) of 2838c3 WT cells subcutaneously injected into naive mice, mice previously immunized with irradiated, dead 2838c3 EV cells (dEV immunization), or mice previously immunized with dead 2838c3 KO-A cells (dKO immunization) (n = 7 mice/group). Data represent mean ± SEM. (E) Vβ repertoire analysis of CD4+ and CD8+ T cells in the spleens of naive mice (n = 5, from A), immunized mice challenged with 2838c3 WT tumor cells day 3 after subcutaneous injection (n = 3), or immunized mice challenged with 2838c3 WT tumor cells on day 10 after subcutaneous injection (n = 3). Data represent mean ± SEM. Statistical analysis by 2-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.