Functional genomics identifies N-acetyllactosamine extension of complex N-glycans as a mechanism to evade lysis by natural killer cells.

Publisher: Cell Press Country of Publication: United States NLM ID: 101573691 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2211-1247 (Electronic) NLM ISO Abbreviation: Cell Rep Subsets: MEDLINE

Original Publication: [Cambridge, MA] : Cell Press, c 2012-

Polysaccharides*/metabolism ; Killer Cells, Natural*/metabolism ; Killer Cells, Natural*/immunology; Humans ; Amino Sugars/metabolism ; Genomics/methods ; Rituximab/pharmacology ; Rituximab/metabolism ; Cell Line, Tumor

Natural killer (NK) cells are primary defenders against cancer precursors, but cancer cells can persist by evading immune surveillance. To investigate the genetic mechanisms underlying this evasion, we perform a genome-wide CRISPR screen using B lymphoblastoid cells. SPPL3, a peptidase that cleaves glycosyltransferases in the Golgi, emerges as a top hit facilitating evasion from NK cytotoxicity. SPPL3-deleted cells accumulate glycosyltransferases and complex N-glycans, disrupting not only binding of ligands to NK receptors but also binding of rituximab, a CD20 antibody approved for treating B cell cancers. Notably, inhibiting N-glycan maturation restores receptor binding and sensitivity to NK cells. A secondary CRISPR screen in SPPL3-deficient cells identifies B3GNT2, a transferase-mediating poly-LacNAc extension, as crucial for resistance. Mass spectrometry confirms enrichment of N-glycans bearing poly-LacNAc upon SPPL3 loss. Collectively, our study shows the essential role of SPPL3 and poly-LacNAc in cancer immune evasion, suggesting a promising target for cancer treatment.
Competing Interests: Declaration of interests The authors declare no competing interests.
(Published by Elsevier Inc.)

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R24 GM137782 United States GM NIGMS NIH HHS; Z01 AI000842 United States ImNIH Intramural NIH HHS

Keywords: B cell cancer; B3GNT2; CP: Cancer; CRISPR screen; N-glycosylation; NK cell; SPPL3; cancer; glycosyltransferase; immunotherapy; natural killer; poly-LacNAc; rituximab

0 (Polysaccharides)
0 (Amino Sugars)
3Y5B2K5OOK (N-acetyllactosamine)
4F4X42SYQ6 (Rituximab)

Date Created: 20240415 Date Completed: 20240426 Latest Revision: 20240614

20240614

PMC11170631

10.1016/j.celrep.2024.114105

38619967