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bioRxiv · 10.1101/2025.08.28.672833

Towards active vaccination against tumour endothelial marker Robo4

Abstract

Targeting tumour antigens is a major challenge in cancer-immunotherapy. We use active vaccination to induce antibodies targeting self-antigen Robo4, which is selectively expressed on tumour vascular endothelium, supporting vascular development. Our previous work showed that a conjugate of Robo4 with a foreign carrier protein can induce autoantibodies specific to Robo4, which inhibited angiogenesis and tumour growth. The current project aims to translate the vaccine protocol to exploit a carrier protein used in routine human vaccination schedules. The well-characterised, non-toxic fragment C of tetanus toxin (TTc) was selected as the carrier protein. Here we show that priming with the carrier TTc followed by boost with Robo4-TTc (R4-TTc) efficiently induces strong antibody responses to Robo4 and inhibits tumour growth in LLC1 and 4T1 tumour models. The growth inhibition was correlated with anti-Robo4 IgG1 titres. Furthermore, we observed decreased vessel formation and increased immune cell infiltration in tumours from R4-TTc vaccinated mice in the absence of detectable adverse effects on health. The data indicate that this vaccination strategy remodels tumour vessels and probably promotes immunogenic pathway activation, therefore repressing tumour growth. One Sentence SummaryA conjugate vaccine inducing antibody responses to tumour endothelial markers Robo4 can inhibit tumour growth.

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BibTeXRIS

Escobar-Riquelme, F. N., Kara, M. A., Price, M., Hidalgo-Gajardo, A., Carr, H., Bending, D., Bicknell, R., Savelyeva, N., Zhang, Y., Toellner, K.-M.. 2025-09-02. Towards active vaccination against tumour endothelial marker Robo4. https://doi.org/10.1101/2025.08.28.672833

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