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2-Photon Characterization of Optical Proteolytic Beacons for Imaging Changes in Matrix-Metalloprotease Activity in a Mouse Model of Aneurysm

Published online by Cambridge University Press:  23 February 2016

Darren G. Haskett ,
David Maestas ,
Stephen J. Howerton ,
Tyler Smith ,
D. Catalina Ardila ,
Tom Doetschman ,
Urs Utzinger ,
Dominic McGrath ,
J. Oliver McIntyre  and
Jonathan P. Vande Geest
Darren G. Haskett
Affiliation:
Graduate Interdisciplinary Program of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA
David Maestas
Affiliation:
Department of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA Department of Molecular and Cellular Biology, The University of Arizona, Tucson, AZ 85721, USA
Stephen J. Howerton
Affiliation:
Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, AZ 85721, USA
Tyler Smith
Affiliation:
Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, AZ 85721, USA
D. Catalina Ardila
Affiliation:
Graduate Interdisciplinary Program of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA
Tom Doetschman
Affiliation:
Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA BIO5 Institute, The University of Arizona, Tucson, AZ 85721, USA
Urs Utzinger
Affiliation:
Graduate Interdisciplinary Program of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA Department of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA BIO5 Institute, The University of Arizona, Tucson, AZ 85721, USA
Dominic McGrath
Affiliation:
Graduate Interdisciplinary Program of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ 85721, USA BIO5 Institute, The University of Arizona, Tucson, AZ 85721, USA
J. Oliver McIntyre
Affiliation:
Departments of Radiology and Radiological Sciences and Cancer Biology, Vanderbilt University, Nashville, TN 37232, USA
Jonathan P. Vande Geest*
Affiliation:
Graduate Interdisciplinary Program of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA Department of Biomedical Engineering, The University of Arizona, Tucson, AZ 85721, USA Department of Aerospace and Mechanical Engineering, The University of Arizona, Tucson, AZ 85721, USA BIO5 Institute, The University of Arizona, Tucson, AZ 85721, USA Department of Bioengineering, The University of Pittsburgh, Pittsburgh, PA 15219, USA
*
*Corresponding author.jpv20@pitt.edu
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Abstract

Abdominal aortic aneurysm is a multifactorial disease that is a leading cause of death in developed countries. Matrix-metalloproteases (MMPs) are part of the disease process, however, assessing their role in disease initiation and progression has been difficult and animal models have become essential. Combining Förster resonance energy transfer (FRET) proteolytic beacons activated in the presence of MMPs with 2-photon microscopy allows for a novel method of evaluating MMP activity within the extracellular matrix (ECM). Single and 2-photon spectra for proteolytic beacons were determined in vitro. Ex vivo experiments using the apolipoprotein E knockout angiotensin II-infused mouse model of aneurysm imaged ECM architecture simultaneously with the MMP-activated FRET beacons. 2-photon spectra of the two-color proteolytic beacons showed peaks for the individual fluorophores that enable imaging of MMP activity through proteolytic cleavage. Ex vivo imaging of the beacons within the ECM revealed both microstructure and MMP activity. 2-photon imaging of the beacons in aneurysmal tissue showed an increase in proteolytic cleavage within the ECM (p<0.001), thus indicating an increase in MMP activity. Our data suggest that FRET-based proteolytic beacons show promise in assessing MMP activity within the ECM and will therefore allow future studies to identify the heterogeneous distribution of simultaneous ECM remodeling and protease activity in aneurysmal disease.

Keywords

AAAMMPApoEFRETmultiphoton
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 22 , Issue 2 , April 2016 , pp. 349 - 360
Copyright
© Microscopy Society of America 2016 

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