GRP receptor-targeted PET of a rat pancreas carcinoma xenograft in nude mice with a 68Ga-labeled bombesin(6-14) analog.
Bombesin (BN), a 14-amino-acid peptide, shows high affinity for the human gastrin-releasing peptide receptor (GRP-r), which is overexpressed on several types of cancer, including prostate, breast, gastrointestinal, and small cell lung cancer. Thus, radiolabeled BN or BN analogs may prove to be specific tracers for diagnostic and therapeutic targeting of GRP-r-positive tumors in nuclear medicine. This study evaluated a novel BN analog labeled with the positron emitter 68Ga for receptor imaging with PET.


METHODS
DOTA-PEG2-[D-Tyr6,beta-Ala11,Thi13,Nle14] BN(6-14) amide (BZH3) (DOTA is 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid; PEG is ethyleneglycol (2-aminoethyl)carboxymethyl ether) was synthetized using the Fmoc strategy and radiolabeled with either 67Ga or 177Lu for in vitro and biodistribution experiments. 68Ga for PET was obtained from a 68Ge/68Ga generator. In vitro binding, internalization, and efflux were determined using the pancreatic tumor cell line AR42J. Biodistribution of the peptide as a function of time and dose was studied in AR42J tumor-bearing mice.


RESULTS
In vitro assays demonstrated a high affinity of 67Ga-BZH3 (dissociation constant = 0.46 nmol/L), a rapid internalization (70% of total cell-associated activity was endocytosed after a 15-min incubation), and an intracellular retention half-life (t1/2) of the 67Ga activity of 16.5 +/- 2.4 h. Biodistribution indicated a dose-dependent uptake in the tumor and a prolonged tumor residence time (t1/2 approximately 16 h). Clearance from GRP-r-negative tissues was fast, resulting in high tumor-to-tissue ratios as early as 1 h after injection. Replacing 67Ga by 177Lu, a therapeutic radionuclide, for peptide labeling resulted in a slightly reduced (approximately 20%) tumor uptake and tumor residence time of 177Lu-BZH3. In contrast, 177Lu decline in the pancreas was significantly accelerated by a factor of 3 compared with that of 67Ga. PET of mice with 68Ga-BZH3 clearly delineated tumors in the mediastinal area.


CONCLUSIONS
The promising in vivo data of 68Ga-BZH3 indicate its potential for an improved localization of GRP-r-positive tumors and also suggest its application in patients. PET may also be favorably used for GRP-r density determination, a prerequisite for therapeutic applications.
Schuhmacher
Jochen
J
Department of Diagnostic and Therapeutic Radiology, German Cancer Research Center, Heidelberg, Germany. j.schuhmacher@dkfz.de
Zhang
Hanwen
H
Doll
Josef
J
Mäcke
Helmut R
HR
Matys
Ronald
R
Hauser
Harald
H
Henze
Marcus
M
Haberkorn
Uwe
U
Eisenhut
Michael
M
D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D019886:Gastrin-Releasing Peptide / Q000378:metabolism*; D008657:Metabolic Clearance Rate; D051379:Mice; D008819:Mice, Nude; D009842:Oligopeptides / Q000493:pharmacokinetics*; D009928:Organ Specificity; D009942:Organometallic Compounds / Q000493:pharmacokinetics*; D010190:Pancreatic Neoplasms / Q000000981:diagnostic imaging* / Q000378:metabolism*; D049268:Positron-Emission Tomography / Q000379:methods*; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics; D051381:Rats; D014018:Tissue Distribution; D014183:Transplantation, Heterologous
C500880:68Ga bombesin(6-14)amide, DOTA-PEG2-Tyr(6)-beta-Ala(11)-Thi(13)-Nle(14)-; D009842:Oligopeptides; D009942:Organometallic Compounds; D019275:Radiopharmaceuticals; D019886:Gastrin-Releasing Peptide