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Selective Imaging of Vascular Endothelial Growth Factor Receptor-1 and Receptor-2 in Atherosclerotic Lesions in Diabetic and Non-diabetic ApoE−/− Mice

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Abstract

Purpose

Plaque vulnerability is associated with inflammation and angiogenesis, processes that rely on vascular endothelial growth factor (VEGF) signaling via two receptors, VEGFR-1 and VEGFR-2. We have recently reported that enhanced uptake of scVEGF-PEG-DOTA/Tc-99m (scV/Tc) single photon emission computed tomography (SPECT) tracer that targets both VEGFR-1 and VEGFR-2, identifies accelerated atherosclerosis in diabetic relative to non-diabetic ApoE−/− mice. Since VEGFR-1 and VEGFR-2 may play different roles in atherosclerotic plaques, we reasoned that selective imaging of each receptor can provide more detailed information on plaque biology.

Procedures

Recently described VEGFR-1 and VEGFR-2 selective mutants of scVEGF, named scVR1 and scVR2, were site-specifically derivatized with Tc-99m chelator DOTA via 3.4 kDa PEG linker, and their selectivity to the cognate receptors was confirmed in vitro. scVR1 and scVR2 conjugates were radiolabeled with Tc-99m to specific activity of 110 ± 11 MBq/nmol, yielding tracers named scVR1/Tc and scVR2/Tc. 34–40 week old diabetic and age-matched non-diabetic ApoE−/− mice were injected with tracers, 2–3 h later injected with x-ray computed tomography (CT) contrast agent and underwent hybrid SPECT/CT imaging. Tracer uptake, localized to proximal aorta and brachiocephalic vessels, was quantified as %ID from. Tracer uptake was also quantified as %ID/g from gamma counting of harvested plaques. Harvested atherosclerotic arterial tissue was used for immunofluorescent analyses of VEGFR-1 and VEGFR-2 and various lineage-specific markers.

Results

Focal, receptor-mediated uptake in proximal aorta and brachiocephalic vessels was detected for both scVR1/Tc and scVR2/Tc tracers. Uptake of scVR1/Tc and scVR2/Tc was efficiently inhibited only by “cold” proteins of the same receptor selectivity. Tracer uptake in this area, expressed as %ID, was higher in diabetic vs. non- diabetic mice for scVR1/Tc (p = 0.01) but not for scVR2/Tc. Immunofluorescent analysis revealed enhanced VEGFR-1 prevalence in and around plaque area in diabetic mice.

Conclusions

Selective VEGFR-1 and VEGFR-2 imaging of atherosclerotic lesions may be useful to explore plaque biology and identify vulnerability.

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Acknowledgments

This work was supported in part by NIH SBIR Phase I contract HHSN26801400041C to JMB and LLJ. We thank Dr. Lucrecia Marques-Rosado for help with immunohistochemistry.

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Authors and Affiliations

  1. Department of Medicine, Columbia University Medical Center, 622 St 168th St, PH 10, room 203, New York, NY, 10032, USA

    Yared Tekabe, Lynne L. Johnson, Krissy Rodriquez & Qing Li

  2. SibTech, Inc., 115A Commerce Drive, Brookfield, CT, 06804, USA

    Marina Backer & Joseph M. Backer

Authors
  1. Yared Tekabe
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  2. Lynne L. Johnson
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  3. Krissy Rodriquez
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  4. Qing Li
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  5. Marina Backer
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Corresponding author

Correspondence to Lynne L. Johnson.

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Conflict of Interest

Joseph Backer is a shareholder in SibTech, Inc.

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Tekabe, Y., Johnson, L.L., Rodriquez, K. et al. Selective Imaging of Vascular Endothelial Growth Factor Receptor-1 and Receptor-2 in Atherosclerotic Lesions in Diabetic and Non-diabetic ApoE−/− Mice. Mol Imaging Biol 20, 85–93 (2018). https://doi.org/10.1007/s11307-017-1045-2

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  • DOI: https://doi.org/10.1007/s11307-017-1045-2

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