N-acetylglucosaminyltransferase-V (GnT-V)-enriched small extracellular vesicles mediate N-glycan remodeling in recipient cells

Tetsuya Hirata; Yoichiro Harada; Koichiro M Hirosawa; Yuko Tokoro; Kenichi G N Suzuki; Yasuhiko Kizuka

doi:10.1016/j.isci.2022.105747

N-acetylglucosaminyltransferase-V (GnT-V)-enriched small extracellular vesicles mediate N-glycan remodeling in recipient cells

iScience. 2022 Dec 6;26(1):105747. doi: 10.1016/j.isci.2022.105747. eCollection 2023 Jan 20.

Authors

Tetsuya Hirata¹, Yoichiro Harada², Koichiro M Hirosawa³, Yuko Tokoro¹, Kenichi G N Suzuki³, Yasuhiko Kizuka¹

Affiliations

¹ Laboratory of Glyco-biochemistry, Institute for Glyco-core Research (iGCORE), Gifu University, Gifu 501-1193, Japan.
² Department of Glyco-Oncology and Medical Biochemistry, Osaka International Cancer Institute, Osaka 541-8567, Japan.
³ Laboratory of Cell Biophysics, Institute for Glyco-core Research (iGCORE), Gifu University, Gifu 501-1193, Japan.

Abstract

Small extracellular vesicles (sEVs) secreted from cancer cells play pivotal roles in cancer metastasis and malignancy by transferring biomolecules and conditioning future metastatic sites. Studies have elucidated structures and functions of glycans on sEVs; however, whether sEVs remodel glycans in recipient cells remains poorly understood. Here, we examined the enzyme activity of glycosyltransferases for complex N-glycan biosynthesis in cancer-derived sEVs and discovered that cancer-related glycosyltransferase, N-acetylglucosaminyltransferase-V (GnT-V, a.k.a. MGAT5), is selectively enriched in sEVs among various glycosyltransferases. GnT-V in sEVs is a cleaved form, and cleavage by SPPL3 protease is necessary for loading GnT-V in sEVs. Fractionation experiments and single-particle imaging further revealed that GnT-V was enriched in non-exosomal sEVs. Strikingly, we found that enzymatically active GnT-V in sEVs was transferred to recipient cells and the N-glycan structures of recipient cells were remodeled to express GnT-V-produced glycans. Our results suggest GnT-V-enriched sEVs' role in glycan remodeling in cancer metastasis.

Keywords: Biochemistry; Cancer; Cell biology.