Glycosylated extracellular vesicles released by glioblastoma cells are decorated by CCL18 allowing for cellular uptake via chemokine receptor CCR8

Abstract

Cancer cells release extracellular vesicles (EVs) that contain functional biomolecules such as RNA and proteins. EVs are transferred to recipient cancer cells and can promote tumour progression and therapy resistance. Through RNAi screening, we identified a novel EV uptake mechanism involving a triple interaction between the chemokine receptor CCR8 on the cells, glycans exposed on EVs and the soluble ligand CCL18. This ligand acts as bridging molecule, connecting EVs to cancer cells. We show that glioblastoma EVs promote cell proliferation and resistance to the alkylating agent temozolomide (TMZ). Using in vitro and in vivo stem-like glioblastoma models, we demonstrate that EV-induced phenotypes are neutralised by a small molecule CCR8 inhibitor, R243. Interference with chemokine receptors may offer therapeutic opportunities against EV-mediated cross-talk in glioblastoma.

Glycan modification of glioblastoma‐derived extracellular vesicles enhances receptor‐mediated targeting of dendritic cells

“…TEM images also confirmed the typical morphology of EVs (Figure 1(b,d)). Size distribution profiles, quantified by measuring the diameter of 100 at random‐selected EVs in eight TEM images (Figure 1(d,e,f)) and by nanoparticle tracking analysis (NTA, Figure 1(g)), were comparable to previously reported size of exosomes/small EVs (70–200 nm) isolated from glioblastoma culture media [2,4,9,57].…”

“…To circumvent the need for personalized neo‐antigen identification and peptide synthesis, we proposed that EVs may represent a more comprehensive, cell‐free, autologous source of neo‐antigens and tumour‐associated antigens, which could be targeted for efficient DC loading [3,35,67]. Since EVs can be internalized by recipient cells via glycan‐specific receptors [4–6], we profiled the surface glycosylation of glioblastoma cell line‐derived EVs and modified their glycosylation for enhanced internalization by DCs. By measuring lectin‐binding properties of EVs in both ELISA‐based assays and immunogold‐TEM, we showed that the surface glycoconjugates of our EV preparations were dominated by immune inhibitory sialic acid‐capped N ‐glycans and complex bi‐antennary glycans.…”

“…All cells including glioblastoma cells produce EVs, thereby releasing small packages of information and delivering them to target cells [1–3]. Although capture and uptake of EVs by recipient cells could occur through membrane fusion or endocytosis, several studies have reported receptor‐mediated EV uptake in a glycan‐dependent way [4–6]. The glycan profile of EVs from different sources (T‐cells, carcinoma and melanoma cell lines and human breast milk) has been studied by lectin microarray and mass spectrometry showing a relatively conserved glycan profile with enrichment of polylactosamine and α2‐6 linked sialic acid residues, complex type N ‐glycans and high mannose structures [7–9].…”

CC Chemokines in a Tumor: A Review of Pro-Cancer and Anti-Cancer Properties of Receptors CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 Ligands

“…In addition, this chemokine participates in the differentiation of immature dendritic cells into tumor-associated dendritic cells (TADC) [ 268 , 269 ]. Other functions of CCL18 in the tumor include participation in the intercellular communication dependent on extracellular vesicles [ 270 ]. This chemokine binds to glycosaminoglycans on extracellular vesicles, which allows them to be retained on cells with an expression of CCR8, a receptor for CCL18.…”

Biological membranes in EV biogenesis, stability, uptake, and cargo transfer: an ISEV position paper arising from the ISEV membranes and EVs workshop

“…Molecules exposed on the EV surface, including fibronectin, PS, and integrins, interact with heparan sulphate proteoglycans [86], T‐cell immunoglobulin‐ and mucin‐domain‐containing molecules [87], and cell‐associated extracellular matrix [8] present on recipient cells. Chemokines and their receptors may act as bridges linking the two membranes together, as they are expressed on both EVs and cells [88,89]. Although follicular dendritic cells do not express MHC‐II, their surface can become fully decorated with vesicles that do (likely derived from B‐cells) [90].…”