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Preclinical Evaluation of virus-like particle Vaccine Against Carbonic Anhydrase IX Efficacy in a Mouse Breast Cancer Model System

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Abstract

Carbonic anhydrase IX (CAIX) is a cancer-associated membrane protein frequently overexpressed in hypoxic solid tumours leading to enhanced tumour cell survival and invasion, and it has been proposed to be an attractive tumour-specific molecule for antibody-mediated targeting. This study aimed to generate a virus-like particle (VLP)-based CAIX vaccine candidate and evaluate its efficacy in a mouse model of breast cancer. The prototype murine vaccine was developed based on the ssRNA bacteriophage Qbeta VLPs with chemically coupled murine CAIX protein catalytic domains on their surfaces. The vaccine was shown to efficiently break the natural B cell tolerance against autologous murine CAIX and to induce high-titre Th1-oriented IgG responses in the BALB/c mice. This vaccine was tested in a therapeutic setting by using a triple-negative breast cancer mouse model system comprising 4T1, 4T1-Car9KI and 4T1-Car9KO cells, the latter representing positive and negative controls for murine CAIX production, respectively. The humoural immune responses induced in tumour-bearing animals were predominantly of Th1-type and higher anti-mCAIXc titres correlated with slower growth and lung metastasis development of 4T1 tumours constitutively expressing mCAIX in vivo in the syngeneic host.

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Data Availability

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Abbreviations

CAIX:

Carbonic anhydrase IX

CMV:

Cytomegalovirus

H&E:

Haematoxylin and eosin

IVC:

Individually ventilated cage

KO:

Knock-out

KI:

Knock-in

mCAIXc:

Catalytic domain of murine carbonic anhydrase IX

ssRNA:

Single-stranded RNA

SPF:

Specific pathogen-free

TAA:

Tumour-associated antigen

TNBC:

Triple-negative breast cancer

VLP:

Virus-like particle

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Acknowledgements

The authors declare no competing interests. We are thankful to Dr. Juris Jansons for his technical support with electron microscopy. This study was supported by the European Regional Development Fund (ERDF) grant number 1.1.1.2/VIAA/4/20/728.

Funding

European Regional Development Fund,1.1.1.2/VIAA/4/20/728, Zane Kalniņa.

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Authors and Affiliations

  1. Latvian Biomedical Research and Study Centre, Ratsupites 1, k-1, Riga, LV-1067, Latvia

    Zane Kalniņa, Ilva Liekniņa, Ināra Akopjana, Andris Kazāks & Kaspars Tārs

  2. Faculty of Biology, University of Latvia, Jelgavas 1, Riga, LV-1004, Latvia

    Zane Kalniņa, Vendija Skeltona & Kaspars Tārs

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  1. Zane Kalniņa
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Contributions

All authors have made substantial contributions to the manuscript as described: conceptualisation: KT and ZK; methodology: ZK, IL, IA and AK; formal analysis and investigation: ZK, IL and VS; funding acquisition: ZK; project administration: ZK; supervision: KT; writing—original draft preparation: ZK and IL; writing—review and editing: all authors; visualisation: ZK.

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Correspondence to Zane Kalniņa.

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Kalniņa, Z., Liekniņa, I., Skeltona, V. et al. Preclinical Evaluation of virus-like particle Vaccine Against Carbonic Anhydrase IX Efficacy in a Mouse Breast Cancer Model System. Mol Biotechnol 66, 1206–1219 (2024). https://doi.org/10.1007/s12033-023-01021-5

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