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CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation

INTRODUCTION: In epithelial cancers, truncated O-glycans, such as the Thomson-nouveau antigen (Tn) and its sialylated form (STn), are upregulated on the cell surface and associated with poor prognosis and immunological escape. Recent studies have shown that these carbohydrate epitopes facilitate can...

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Autores principales: Rømer, Troels Boldt, Khoder-Agha, Fawzi, Aasted, Mikkel Koed Møller, de Haan, Noortje, Horn, Sabrina, Dylander, August, Zhang, Tao, Pallesen, Emil Marek Heymans, Dabelsteen, Sally, Wuhrer, Manfred, Høgsbro, Christine Flodgaard, Thomsen, Emil Aagaard, Mikkelsen, Jacob Giehm, Wandall, Hans H
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10627246/
https://www.ncbi.nlm.nih.gov/pubmed/36648436
http://dx.doi.org/10.1093/glycob/cwad003
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author Rømer, Troels Boldt
Khoder-Agha, Fawzi
Aasted, Mikkel Koed Møller
de Haan, Noortje
Horn, Sabrina
Dylander, August
Zhang, Tao
Pallesen, Emil Marek Heymans
Dabelsteen, Sally
Wuhrer, Manfred
Høgsbro, Christine Flodgaard
Thomsen, Emil Aagaard
Mikkelsen, Jacob Giehm
Wandall, Hans H
author_facet Rømer, Troels Boldt
Khoder-Agha, Fawzi
Aasted, Mikkel Koed Møller
de Haan, Noortje
Horn, Sabrina
Dylander, August
Zhang, Tao
Pallesen, Emil Marek Heymans
Dabelsteen, Sally
Wuhrer, Manfred
Høgsbro, Christine Flodgaard
Thomsen, Emil Aagaard
Mikkelsen, Jacob Giehm
Wandall, Hans H
author_sort Rømer, Troels Boldt
collection PubMed
description INTRODUCTION: In epithelial cancers, truncated O-glycans, such as the Thomson-nouveau antigen (Tn) and its sialylated form (STn), are upregulated on the cell surface and associated with poor prognosis and immunological escape. Recent studies have shown that these carbohydrate epitopes facilitate cancer development and can be targeted therapeutically; however, the mechanism underpinning their expression remains unclear. METHODS: To identify genes directly influencing the expression of cancer-associated O-glycans, we conducted an unbiased, positive-selection, whole-genome CRISPR knockout-screen using monoclonal antibodies against Tn and STn. RESULTS AND CONCLUSIONS: We show that knockout of the Zn(2+)-transporter SLC39A9 (ZIP9), alongside the well-described targets C1GALT1 (C1GalT1) and its molecular chaperone, C1GALT1C1 (COSMC), results in surface-expression of cancer-associated O-glycans. No other gene perturbations were found to reliably induce O-glycan truncation. We furthermore show that ZIP9 knockout affects N-linked glycosylation, resulting in upregulation of oligo-mannose, hybrid-type, and α2,6-sialylated structures as well as downregulation of tri- and tetra-antennary structures. Finally, we demonstrate that accumulation of Zn(2+) in the secretory pathway coincides with cell-surface presentation of truncated O-glycans in cancer tissue, and that over-expression of COSMC mitigates such changes. Collectively, the findings show that dysregulation of ZIP9 and Zn(2+) induces cancer-like glycosylation on the cell surface by affecting the glycosylation machinery.
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spelling pubmed-106272462023-11-07 CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation Rømer, Troels Boldt Khoder-Agha, Fawzi Aasted, Mikkel Koed Møller de Haan, Noortje Horn, Sabrina Dylander, August Zhang, Tao Pallesen, Emil Marek Heymans Dabelsteen, Sally Wuhrer, Manfred Høgsbro, Christine Flodgaard Thomsen, Emil Aagaard Mikkelsen, Jacob Giehm Wandall, Hans H Glycobiology Original Article INTRODUCTION: In epithelial cancers, truncated O-glycans, such as the Thomson-nouveau antigen (Tn) and its sialylated form (STn), are upregulated on the cell surface and associated with poor prognosis and immunological escape. Recent studies have shown that these carbohydrate epitopes facilitate cancer development and can be targeted therapeutically; however, the mechanism underpinning their expression remains unclear. METHODS: To identify genes directly influencing the expression of cancer-associated O-glycans, we conducted an unbiased, positive-selection, whole-genome CRISPR knockout-screen using monoclonal antibodies against Tn and STn. RESULTS AND CONCLUSIONS: We show that knockout of the Zn(2+)-transporter SLC39A9 (ZIP9), alongside the well-described targets C1GALT1 (C1GalT1) and its molecular chaperone, C1GALT1C1 (COSMC), results in surface-expression of cancer-associated O-glycans. No other gene perturbations were found to reliably induce O-glycan truncation. We furthermore show that ZIP9 knockout affects N-linked glycosylation, resulting in upregulation of oligo-mannose, hybrid-type, and α2,6-sialylated structures as well as downregulation of tri- and tetra-antennary structures. Finally, we demonstrate that accumulation of Zn(2+) in the secretory pathway coincides with cell-surface presentation of truncated O-glycans in cancer tissue, and that over-expression of COSMC mitigates such changes. Collectively, the findings show that dysregulation of ZIP9 and Zn(2+) induces cancer-like glycosylation on the cell surface by affecting the glycosylation machinery. Oxford University Press 2023-01-16 /pmc/articles/PMC10627246/ /pubmed/36648436 http://dx.doi.org/10.1093/glycob/cwad003 Text en © The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Original Article
Rømer, Troels Boldt
Khoder-Agha, Fawzi
Aasted, Mikkel Koed Møller
de Haan, Noortje
Horn, Sabrina
Dylander, August
Zhang, Tao
Pallesen, Emil Marek Heymans
Dabelsteen, Sally
Wuhrer, Manfred
Høgsbro, Christine Flodgaard
Thomsen, Emil Aagaard
Mikkelsen, Jacob Giehm
Wandall, Hans H
CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation
title CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation
title_full CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation
title_fullStr CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation
title_full_unstemmed CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation
title_short CRISPR-screen identifies ZIP9 and dysregulated Zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation
title_sort crispr-screen identifies zip9 and dysregulated zn(2+) homeostasis as a cause of cancer-associated changes in glycosylation
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10627246/
https://www.ncbi.nlm.nih.gov/pubmed/36648436
http://dx.doi.org/10.1093/glycob/cwad003
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