title: Detecting in vivo urokinase plasminogen activator activity with a catalyCEST MRI contrast agent.
abstract: Urokinase plasminogen activator (uPA) promotes tumor invasion and metastasis. The monitoring of uPA activity using molecular imaging may have prognostic value and be predictive for response to anti-cancer therapies. However, the detection of in vivo enzyme activity with molecular imaging remains a challenge. To address this problem, we designed a nonmetallic contrast agent, GR-4Am-SA, that can be detected with chemical exchange saturation transfer (CEST) MRI. This agent has a peptide that is cleaved by uPA, which causes a CEST signal at 5.0 ppm to decrease, and also has a salicylic acid moiety that can produce a CEST signal at 9.5 ppm, which is largely unresponsive to enzyme activity. The two CEST signals were used to determine a reaction coordinate, representing the extent of enzyme-catalyzed cleavage of the GR-4Am-SA agent during an experimental study. Initial biochemical studies showed that GR-4Am-SA could detect uPA activity in reducing conditions. Subsequently, we used our catalyCEST MRI protocol with the agent to detect the uPA catalysis of GR-4Am-SA in a flank xenograft model of Capan-2 pancreatic cancer. The results showed an average reaction coordinate of 80% ± 8%, which was strongly dependent on the CEST signal at 5.0 ppm. The relative independence of the reaction coordinate on the CEST signal at 9.5 ppm showed that the detection of enzyme activity was largely independent of the concentration of GR-4Am-SA within the tumor tissue. These results demonstrated the advantages of a single CEST agent with biomarker-responsive and unresponsive signals for reliably assessing enzyme activity during in vivo cancer studies.
authors.affiliation: Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ, USA.
authors.forename: Sanhita
authors.initials: S
authors.lastname: Sinharay
authors.affiliation: Department of Medical Imaging, University of Arizona, Tucson, AZ, USA.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Howison
authors.affiliation: University of Arizona Cancer Center, University of Arizona, Tucson, AZ, USA.
authors.forename: Amanda F
authors.initials: AF
authors.lastname: Baker
authors.affiliation: Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ, USA.|Department of Medical Imaging, University of Arizona, Tucson, AZ, USA.|University of Arizona Cancer Center, University of Arizona, Tucson, AZ, USA.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Pagel
mesh_terms: D000465:Algorithms; D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism*; D045744:Cell Line, Tumor; D003287:Contrast Media / Q000493:pharmacokinetics; D004789:Enzyme Activation; D005260:Female; D008279:Magnetic Resonance Imaging / Q000379:methods*; D051379:Mice; D008819:Mice, Nude; D057054:Molecular Imaging / Q000379:methods*; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D066244:Proton Magnetic Resonance Spectroscopy / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
chemical_list: D014408:Biomarkers, Tumor; D003287:Contrast Media; D014568:Urokinase-Type Plasminogen Activator
keywords: CEST MRI; enzyme activity; pancreatic cancer; urokinase plasminogen activator