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bioRxiv · 10.1101/2024.10.26.620411

Gb3 trisaccharide-bearing exosomes as a novel neutralizer for Shiga toxin type 1

Mikolajczyk, K.
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Abstract

Shiga toxin types 1 (Stx1) and 2 (Stx2), produced by Shiga toxin-producing Escherichia coli (STEC) and Shigella dysenteriae, are key virulence factors responsible for severe foodborne diseases, such as hemorrhagic colitis and hemolytic uremic syndrome (HUS). The receptors for Stxs are Gb3 and P1 glycotope, which contain the Gal1[->]4Gal epitope and are synthesized by human 1,4-galactosyltransferase (A4galt). Stx-related infections pose a global public health challenge, owing to the limited therapeutic options due to the restricted use of antibiotics. Therefore, there is an urgent need to develop novel therapeutic strategies. This study proposes an innovative strategy utilizing exosomes derived from CHO-Lec2 cells, which were modified with Functional-Spacer-Lipid (FSL) conjugates bearing the Gb3 carbohydrate epitope (exo-Gb3-FSL). Flow cytometry analysis confirmed the presence of Gal1[->]4Gal disaccharides on exo-Gb3-FSL constructs, enabling them to bind Stx1. Moreover, using CHO-Lec2 cells evaluated the ability of exo-Gb3-FSL agents to bind Stx1 and protect these cells from Stx1-mediated cytotoxicity. For Stx1-treated CHO-Lec2 cells, increased cell survival was observed when using 25 {micro}M exo-Gb3-FSL constructs, compared to control cells. These findings highlight the potential of exosome-based anti-Stx1 agents as promising alternatives to conventional therapies. This innovative strategy may provide novel directions for studies on Stx1 neutralization, offering a valuable strategy for the treatment of Stx-related diseases. Highlights- Exo-Gb3-FSL constructs hold Stx1 binding Gal1[->]4Gal epitope. - Stx1 was effectively bound by exo-Gb3-FSL constructs. - Exo-Gb3-FSL protected CHO-Lec2 cells from Stx1. - Exo-Gb3-FSL is a promising anti-Stx1 agent.

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Mikolajczyk, K.. 2024-10-26. Gb3 trisaccharide-bearing exosomes as a novel neutralizer for Shiga toxin type 1. https://doi.org/10.1101/2024.10.26.620411

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