Integrated Functional Screens and Multi-omics Analyses Identify α-2,3-sialylation As Essential for Melanoma Maintenance

Overview

Journal bioRxiv

Date 2024 Apr 1

PMID 38559078

Authors

Praveen Agrawal

Shuhui Chen

Ana de Pablos

Faezeh Jame-Chenarboo

Eleazar Miera Saenz de Vega

Farbod Darvishian

Iman Osman

Amaia Lujambio

Lara K Mahal

Eva Hernando

Affiliations

Soon will be listed here.

Abstract

Glycosylation is a hallmark of cancer biology, and altered glycosylation influences multiple facets of melanoma growth and progression. To identify glycosyltransferases, glycans, and glycoproteins essential for melanoma maintenance, we conducted an growth screen with a pooled shRNA library of glycosyltransferases, lectin microarray profiling of benign nevi and melanoma patient samples, and mass spectrometry-based glycoproteomics. We found that α-2,3 sialyltransferases ST3GAL1 and ST3GAL2 and corresponding α-2,3-linked sialosides are upregulated in melanoma compared to nevi and are essential for melanoma growth and . Glycoproteomics revealed that glycoprotein targets of ST3GAL1 and ST3GAL2 are enriched in transmembrane proteins involved in growth signaling, including the amino acid transporter Solute Carrier Family 3 Member 2 (SLC3A2/CD98hc). CD98hc suppression mimicked the effect of ST3GAL1 and ST3GAL2 silencing, inhibiting melanoma cell proliferation. We found that both CD98hc protein stability and its pro-survival effect in melanoma are dependent upon α-2,3 sialylation mediated by ST3GAL1 and ST3GAL2. In summary, our studies reveal that α-2,3-sialosides functionally contribute to melanoma maintenance, supporting ST3GAL1 and ST3GAL2 as novel therapeutic targets in these tumors.

References

Elder D . Precursors to melanoma and their mimics: nevi of special sites. Mod Pathol. 2006; 19 Suppl 2:S4-20. DOI: 10.1038/modpathol.3800515. View

Agrawal P, Kurcon T, Pilobello K, Rakus J, Koppolu S, Liu Z . Mapping posttranscriptional regulation of the human glycome uncovers microRNA defining the glycocode. Proc Natl Acad Sci U S A. 2014; 111(11):4338-43. PMC: 3964104. DOI: 10.1073/pnas.1321524111. View

Tian S, Peng P, Li J, Deng H, Zhan N, Zeng Z . SERPINH1 regulates EMT and gastric cancer metastasis via the Wnt/β-catenin signaling pathway. Aging (Albany NY). 2020; 12(4):3574-3593. PMC: 7066881. DOI: 10.18632/aging.102831. View

Aloia A, Petrova E, Tomiuk S, Bissels U, Deas O, Saini M . The sialyl-glycolipid stage-specific embryonic antigen 4 marks a subpopulation of chemotherapy-resistant breast cancer cells with mesenchymal features. Breast Cancer Res. 2015; 17(1):146. PMC: 4660783. DOI: 10.1186/s13058-015-0652-6. View

Li H, Al-Japairai K, Tao Y, Xiang Z . RPN2 promotes colorectal cancer cell proliferation through modulating the glycosylation status of EGFR. Oncotarget. 2017; 8(42):72633-72651. PMC: 5641158. DOI: 10.18632/oncotarget.20005. View

Hillman-Jackson J, Clements D, Blankenberg D, Taylor J, Nekrutenko A, Team G . Using Galaxy to perform large-scale interactive data analyses. Curr Protoc Bioinformatics. 2012; Chapter 10:10.5.1-10.5.47. PMC: 4282168. DOI: 10.1002/0471250953.bi1005s38. View

Chandrasekaran E, Xue J, Xia J, Locke R, Patil S, Neelamegham S . Mammalian sialyltransferase ST3Gal-II: its exchange sialylation catalytic properties allow labeling of sialyl residues in mucin-type sialylated glycoproteins and specific gangliosides. Biochemistry. 2011; 50(44):9475-87. PMC: 3206213. DOI: 10.1021/bi200301w. View

Kleffman K, Levinson G, Rose I, Blumenberg L, Shadaloey S, Dhabaria A . Melanoma-Secreted Amyloid Beta Suppresses Neuroinflammation and Promotes Brain Metastasis. Cancer Discov. 2022; 12(5):1314-1335. PMC: 9069488. DOI: 10.1158/2159-8290.CD-21-1006. View

Joshi H, Hansen L, Narimatsu Y, Freeze H, Henrissat B, Bennett E . Glycosyltransferase genes that cause monogenic congenital disorders of glycosylation are distinct from glycosyltransferase genes associated with complex diseases. Glycobiology. 2018; 28(5):284-294. PMC: 6279177. DOI: 10.1093/glycob/cwy015. View

10.

Bensing B, Li Q, Park D, Lebrilla C, Sullam P . Streptococcal Siglec-like adhesins recognize different subsets of human plasma glycoproteins: implications for infective endocarditis. Glycobiology. 2018; 28(8):601-611. PMC: 6054165. DOI: 10.1093/glycob/cwy052. View

11.

Sturgill E, Aoki K, Lopez P, Colacurcio D, Vajn K, Lorenzini I . Biosynthesis of the major brain gangliosides GD1a and GT1b. Glycobiology. 2012; 22(10):1289-301. PMC: 3425327. DOI: 10.1093/glycob/cws103. View

12.

Kimura R, Yoshimaru T, Matsushita Y, Matsuo T, Ono M, Park J . The GALNT6‑LGALS3BP axis promotes breast cancer cell growth. Int J Oncol. 2020; 56(2):581-595. DOI: 10.3892/ijo.2019.4941. View

13.

Shain A, Bastian B . From melanocytes to melanomas. Nat Rev Cancer. 2016; 16(6):345-58. DOI: 10.1038/nrc.2016.37. View

14.

Lopez A, Cacoub P, Macdougall I, Peyrin-Biroulet L . Iron deficiency anaemia. Lancet. 2015; 387(10021):907-16. DOI: 10.1016/S0140-6736(15)60865-0. View

15.

Krishnan V, Bane S, Kawle P, Naresh K, Kalraiya R . Altered melanoma cell surface glycosylation mediates organ specific adhesion and metastasis via lectin receptors on the lung vascular endothelium. Clin Exp Metastasis. 2005; 22(1):11-24. DOI: 10.1007/s10585-005-2036-2. View

16.

Kabbarah O, Nogueira C, Feng B, Nazarian R, Bosenberg M, Wu M . Integrative genome comparison of primary and metastatic melanomas. PLoS One. 2010; 5(5):e10770. PMC: 2875381. DOI: 10.1371/journal.pone.0010770. View

17.

Rodrigues E, Macauley M . Hypersialylation in Cancer: Modulation of Inflammation and Therapeutic Opportunities. Cancers (Basel). 2018; 10(6). PMC: 6025361. DOI: 10.3390/cancers10060207. View

18.

Orr S, Le D, Long J, Sobieszczuk P, Ma B, Tian H . A phenotype survey of 36 mutant mouse strains with gene-targeted defects in glycosyltransferases or glycan-binding proteins. Glycobiology. 2012; 23(3):363-80. PMC: 3605971. DOI: 10.1093/glycob/cws150. View

19.

Munkley J, Elliott D . Hallmarks of glycosylation in cancer. Oncotarget. 2016; 7(23):35478-89. PMC: 5085245. DOI: 10.18632/oncotarget.8155. View

20.

Doench J, Fusi N, Sullender M, Hegde M, Vaimberg E, Donovan K . Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nat Biotechnol. 2016; 34(2):184-191. PMC: 4744125. DOI: 10.1038/nbt.3437. View