PET imaging of apoptosis with (64)Cu-labeled streptavidin following pretargeting of phosphatidylserine with biotinylated annexin-V.
OBJECTIVE
In vivo detection of apoptosis is a diagnostic tool with potential clinical applications in cardiology and oncology. Radiolabeled annexin-V (anxV) is an ideal probe for in vivo apoptosis detection owing to its strong affinity for phosphatidylserine (PS), the molecular flag on the surface of apoptotic cells. Most clinical studies performed to visualize apoptosis have used (99m)Tc-anxV; however, its poor distribution profile often compromises image quality. In this study, tumor apoptosis after therapy was visualized by positron emission tomography (PET) using (64)Cu-labeled streptavidin (SAv), following pre-targeting of apoptotic cells with biotinylated anxV.


METHODS
Apoptosis was induced in tumor-bearing mice by photodynamic therapy (PDT) using phthalocyanine dyes as photosensitizers, and red light. After PDT, mice were injected i.v. with biotinylated anxV, followed 2 h later by an avidin chase, and after another 2 h with (64)Cu-DOTA-biotin-SAv. PET images were subsequently recorded up to 13 h after PDT.


RESULTS
PET images delineated apoptosis in treated tumors as early as 30 min after (64)Cu-DOTA-biotin-SAv administration, with tumor-to-background ratios reaching a maximum at 3 h post-injection, i.e., 7 h post-PDT. Omitting the administration of biotinylated anxV or the avidin chase failed to provide a clear PET image, confirming that all three steps are essential for adequate visualization of apoptosis. Furthermore, differences in action mechanisms between photosensitizers that target tumor cells directly or via initial vascular stasis were clearly recognized through differences in tracer uptake patterns detecting early or delayed apoptosis.


CONCLUSIONS
This study demonstrates the efficacy of a three-step (64)Cu pretargeting procedure for PET imaging of apoptosis. Our data also confirm the usefulness of small animal PET to evaluate cancer treatment protocols.
Cauchon
Nicole
N
Sherbrooke Molecular Imaging Centre and Department of Nuclear Medicine and Radiobiology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC, Canada.
Langlois
Réjean
R
Rousseau
Jacques A
JA
Tessier
Guillaume
G
Cadorette
Jules
J
Lecomte
Roger
R
Hunting
Darel J
DJ
Pavan
Roberto A
RA
Zeisler
Stefan K
SK
van Lier
Johan E
JE
D000818:Animals; D017304:Annexin A5 / Q000493:pharmacokinetics*; D017209:Apoptosis; D001710:Biotin / Q000493:pharmacokinetics; D045744:Cell Line, Tumor; D003301:Copper Radioisotopes / Q000493:pharmacokinetics; D016503:Drug Delivery Systems / Q000379:methods; D005260:Female; D008657:Metabolic Clearance Rate; D051379:Mice; D008807:Mice, Inbred BALB C; D009374:Neoplasms, Experimental / Q000000981:diagnostic imaging / Q000378:metabolism*; D009928:Organ Specificity; D010718:Phosphatidylserines / Q000378:metabolism*; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics; D019809:Streptavidin / Q000493:pharmacokinetics*; D014018:Tissue Distribution
D017304:Annexin A5; D003301:Copper Radioisotopes; D010718:Phosphatidylserines; D019275:Radiopharmaceuticals; D001710:Biotin; D019809:Streptavidin