Imaging of myeloperoxidase in mice by using novel amplifiable paramagnetic substrates.
OBJECTIVE
To evaluate whether contrast agents for molecular magnetic resonance (MR) imaging can demonstrate the in vivo activity of myeloperoxidase, an enzyme that is secreted by stimulated polymorphonuclear leukocytes, monocytes, and macrophages during inflammation.


METHODS
Animal experiments were approved by the animal care committee. Protocols for the procurement and use of human blood were approved by the institutional review board. Informed consent was obtained from each donor, and HIPAA guidelines were followed for humans. Two paramagnetic myeloperoxidase substrates--that is, gadolinium-5-hydroxytryptamide-tetraazacyclododecane tetraacetic acid (Gd-5-HT-DOTA) and Gd-bis-5-HT-diethylenetriaminepentaacetic acid (Gd-bis-5-HT-DTPA)--were synthesized. Indium 111-labeled bis-5-HT-DTPA was used to determine biodistribution and target localization. A total of 22 mice were used in three models. In the first model, human myeloperoxidase was embedded in a basement membrane matrix gel and was injected intramuscularly. In the second model, lipopolysaccharide (LPS) from Escherichia coli was embedded in a basement membrane matrix gel and was injected intramuscularly to induce endogenous myeloperoxidase secretion. In the third model, LPS was injected intramuscularly to induce myositis. Statistical significance was calculated for contrast-to-noise ratio (CNR) curves by using the Kolmogorov-Smirnov test.


RESULTS
After the administration of Gd-bis-5-HT-DTPA, strong MR signal enhancement (up to 2.5-fold increase in CNR, P < .001) was observed in vivo for implants that contained human myeloperoxidase. In the LPS-induced myositis model, a smaller visible difference was seen (1.3-fold increase in CNR, P < .001), which was consistent with the fact that endogenous mouse myeloperoxidase is only about 10%-20% as active as human myeloperoxidase. Prolonged contrast material enhancement was observed in the myeloperoxidase-containing areas that were injected with Gd-5-HT-DOTA or Gd-bis-5-HT-DTPA but was not observed in areas that were injected with Gd-DTPA or Gd-dopamine-DOTA (P < .05). Single photon emission computed tomography combined with computed tomography was used to confirm the increased retention of contrast agents at sites that contained human myeloperoxidase, and the results of biodistribution studies demonstrated a more than fourfold increase radiotracer accumulation at these sites.


CONCLUSIONS
Human and mouse myeloperoxidase activity in myeloperoxidase implants and inflamed tissues can be visualized and reported in vivo by using myeloperoxidase-sensitive "smart" molecular imaging probes.
Chen
John W
JW
Center for Molecular Imaging Research, Massachusetts General Hospital, Harvard Medical School, 5404 Building 149, 13th St, Charlestown, MA 02129, USA. chenjo@helix.mgh.harvard.edu
Querol Sans
Manel
M
Bogdanov
Alexei
A
Weissleder
Ralph
R
D000818:Animals; D003287:Contrast Media / Q000138:chemical synthesis*; D006571:Heterocyclic Compounds; D006801:Humans; D007273:Injections, Intramuscular; D008070:Lipopolysaccharides; D051379:Mice; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009942:Organometallic Compounds / Q000138:chemical synthesis*; D009195:Peroxidase / Q000378:metabolism*; D018709:Statistics, Nonparametric
D003287:Contrast Media; D006571:Heterocyclic Compounds; D008070:Lipopolysaccharides; D009942:Organometallic Compounds; C496989:glycylmethyl DOTA(Gd) 5-hydroxytryptamide; C050823:gadolinium 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetate; D009195:Peroxidase