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Specific Targeting of Atherosclerotic Plaques in ApoE−/− Mice Using a New Camelid sdAb Binding the Vulnerable Plaque Marker LOX-1

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Abstract

Purpose

Molecular imaging has the potential to provide quantitative information about specific biological aspects of developing atherosclerotic lesions. This requires the generation of reliable, highly specific plaque tracers. This study reports a new camelid single-domain antibody fragment (sdAb) targeting the Lectin-like oxidized low-density lipoprotein receptor (LOX-1), a biomarker for the detection and molecular phenotyping of vulnerable atherosclerotic plaques.

Procedures

A camelid sdAb was generated and selected for high affinity binding to LOX-1. Ex vivo biodistribution and in vivo single photon emission computed tomography (SPECT)/computed tomography (CT) imaging studies were performed in wild-type mice and in fat-fed atherosclerotic apolipoprotein E-deficient mice with 99mTc-labeled sdAbs. Gamma-counting and autoradiography analyses were performed on dissected aorta segments with different degrees of plaque burden. The specificity of the LOX-1-targeting sdAb was evaluated by blocking with unlabeled sdAb or by comparison with a nontargeting 99mTc-labeled control sdAb.

Results

We generated a sdAb binding LOX-1 with a KD of 280 pM ± 62 pM affinity. After 99mTc-labeling, the tracer had radiochemical purity higher then 99 % and retained specificity in in vitro binding studies. Tracer blood clearance was fast with concomitant high kidney retention. At 3 h after injection, uptake in tissues other than plaques was low and not different than background, suggesting a restricted expression pattern of LOX-1. Conversely, uptake in aortic segments increased with plaque content and was due to specific LOX-1 binding. In vivo SPECT/CT imaging 160 min after injection in atherosclerotic mice confirmed specific targeting of LOX-1-expressing aortic plaques.

Conclusions

The LOX-sdAb specifically targets LOX-1-expressing atherosclerotic plaques within hours after injection. The possibility to image LOX-1 rapidly after administration combined with the favourable biodistribution of a sdAb are beneficial for molecular phenotyping of atherosclerotic plaques and the generation of a future prognostic tracer.

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Acknowledgments

We thank Cindy Peleman, Isabel Remory and Emmy De Blay for their technical assistance. Jens De Vos and Sam Massa have a PhD fellowship of the Research Foundation Flanders (FWO). Tony Lahoutte is a Senior Clinical Investigator of the Research Foundation Flanders (FWO). The research at ICMI is funded by the Belgian State, Nationaal Kankerplan, Vlaamse liga tegen kanker, Stichting tegen kanker, FWO-Vlaanderen, IWT and Vrije Universiteit Brussel.

Conflict of Interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Laboratory of Cellular and Molecular Immunology (CMIM), Vrije Universiteit Brussel (VUB), Pleinlaan 2, Brussels, 1050, Belgium

    Jens De Vos, Sam Massa, Serge Muyldermans & Nick Devoogdt

  2. In vivo Cellular and Molecular Imaging Laboratory (ICMI), Vrije Universiteit Brussel (VUB), Brussels, Belgium

    Jens De Vos, Catarina Xavier, Sam Massa, Tony Lahoutte & Nick Devoogdt

  3. Department of Structural Biology, Vlaams Instituut voor Biotechnologie (VIB), Pleinlaan 2, Brussels, 1050, Belgium

    Jens De Vos, Sam Massa & Serge Muyldermans

  4. Cell Differentiation Lab, Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, Brussels, 1090, Belgium

    Iris Mathijs & Luc Bouwens

  5. Central Veterinary Research Laboratory, Dubai, United Arab Emirates

    Ulrich Wernery

  6. Nuclear Medicine Department, UZ Brussel, Laarbeeklaan 101, Brussels, 1090, Belgium

    Tony Lahoutte

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  1. Jens De Vos
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Correspondence to Nick Devoogdt.

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De Vos, J., Mathijs, I., Xavier, C. et al. Specific Targeting of Atherosclerotic Plaques in ApoE−/− Mice Using a New Camelid sdAb Binding the Vulnerable Plaque Marker LOX-1. Mol Imaging Biol 16, 690–698 (2014). https://doi.org/10.1007/s11307-014-0731-6

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  • DOI: https://doi.org/10.1007/s11307-014-0731-6

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