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Selective Imaging of Matrix Metalloproteinase-13 to Detect Extracellular Matrix Remodeling in Atherosclerotic Lesions

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Abstract

Purpose

Overexpression and activation of matrix metalloproteinase-13 (MMP-13) within atheroma increases susceptibility to plaque rupture, a major cause of severe cardiovascular complications. In comparison to pan-MMP targeting [18F]BR-351, we evaluated the potential for [18F]FMBP, a selective PET radiotracer for MMP-13, to detect extracellular matrix (ECM) remodeling in vascular plaques possessing markers of inflammation.

Procedures

[18F]FMBP and [18F]BR-351 were initially assessed in vitro by incubation with en face aortae from 8 month-old atherogenic ApoE−/− mice. Ex vivo biodistributions, plasma metabolite analyses, and ex vivo autoradiography were analogously performed 30 min after intravenous radiotracer administration in age-matched C57Bl/6 and ApoE−/− mice under baseline or homologous blocking conditions. En face aortae were subsequently stained with Oil Red O (ORO), sectioned, and subject to immunofluorescence staining for Mac-2 and MMP-13.

Results

High-resolution autoradiographic image analysis demonstrated target specificity and regional concordance to lipid-rich lesions. Biodistribution studies revealed hepatobiliary excretion, low accumulation of radioactivity in non-excretory organs, and few differences between strains and conditions in non-target organs. Plasma metabolite analyses uncovered that [18F]FMBP exhibited excellent in vivo stability (≥74% intact) while [18F]BR-351 was extensively metabolized (≤37% intact). Ex vivo autoradiography and histology of en face aortae revealed that [18F]FMBP, relative to [18F]BR-351, exhibited 2.9-fold greater lesion uptake, substantial specific binding (68%), and improved sensitivity to atherosclerotic tissue (2.9-fold vs 2.1-fold). Immunofluorescent staining of aortic en face cross sections demonstrated elevated Mac-2 and MMP-13-positive areas within atherosclerotic lesions identified by [18F]FMBP ex vivo autoradiography.

Conclusions

While both radiotracers successfully identified atherosclerotic plaques, [18F]FMBP showed superior specificity and sensitivity for lesions possessing features of destructive plaque remodeling. The detection of ECM remodeling by selective targeting of MMP-13 may enable characterization of high-risk atherosclerosis featuring elevated collagenase activity.

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Acknowledgements

We are grateful to Dr. Rob Beanlands and Dr. Katey Rayner for illuminating discussions, Dr. Tayebeh Hadizad and Daniel Duan for isotope production, and the University of Ottawa Heart Institute Animal Care and Veterinary Services for their contributions to this work.

Funding

The authors receive financial support from CIHR Project Grant 366633, CFI JELF 36848, Ontario Ministry of Research Innovation and Science ER17-13-119, and the Faculty of Medicine and Division of Cardiology at University of Ottawa. A.B. was supported by NSERC USRA and OGS. G.F. was supported by CIHR CGSM and QEII-GSST.

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Authors and Affiliations

  1. Department of Chemistry and Biomolecular Sciences, University of Ottawa, Ottawa, Ontario, K1N 6N5, Canada

    Ariel Buchler & Benjamin H. Rotstein

  2. University of Ottawa Heart Institute, Ottawa, Ontario, K1Y 4W7, Canada

    Ariel Buchler, Maxime Munch, Gedaliah Farber, Xiaoling Zhao, Rami Al-Haddad, Eadan Farber & Benjamin H. Rotstein

  3. Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario, K1H 8M5, Canada

    Maxime Munch, Gedaliah Farber, Rami Al-Haddad & Benjamin H. Rotstein

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  1. Ariel Buchler
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Contributions

Project design—A.B. and B.H.R.

Method development—A.B., M.M., G.F., X.Z., and R.A-H.

Data acquisition and analysis—A.B., M.M., G.F., and E.F.

Writing and revision of the manuscript—A.B., M.M., G.F., and B.H.R.

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Correspondence to Benjamin H. Rotstein.

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Buchler, A., Munch, M., Farber, G. et al. Selective Imaging of Matrix Metalloproteinase-13 to Detect Extracellular Matrix Remodeling in Atherosclerotic Lesions. Mol Imaging Biol 24, 93–103 (2022). https://doi.org/10.1007/s11307-021-01626-9

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