Pretargeting in tumored mice with radiolabeled morpholino oligomer showing low kidney uptake
We have recently shown that accumulation in mouse kidneys of technetium-99m labeled phosphorodiamidate morpholinos (MORFs) increases with the number of cytosines in the base sequence. To improve tumor/kidney ratios in tumored mice, pretargeting studies were performed with a cytosine-free MORF. An 18-mer MORF (5′-TCTTCTACTTCACAACTA) was conjugated to the anti-CEA antibody MN14 (Immunomedics) and administered to nude mice bearing LS174T tumors. Thereafter, the 99mTc-labeled cytosine-free cMORF (5′-TAGTTGTGAAGTAGAAGA-amide-MAG3) was administered. For comparison, the identical study was repeated but with our original pair of 18-mer MORFs (5′-GGGTGTACGTCACAACTA-conjugated MN14 and 99mTc-labeled 5′-TAGTTGTGACGTACACCC-amide-MAG3). Surface plasmon resonance was used to show that the hybridization affinities of the original and the modified pair of MORFs were essentially equal. Hybridization of the cytosine-free cMORF-99mTc to MN14-MORF was demonstrated in vitro by size-exclusion high-performance liquid chromatography. At 3 h, kidney levels in normal mice were 2.0%ID/organ for the modified cMORF vs. 4.1%ID/organ for the original cMORF sequence, while at 24 h, these values were 0.9% vs 1.8%ID/organ. Pretargeting studies in tumored mice receiving 25 μg of conjugated antibody, 0.5 μg of labeled cMORF 48 h later, followed by imaging and sacrifice at 3 h showed that kidney levels were reduced using the cytosine-free cMORF. Moreover, tumor accumulation was about 3.6%ID/g and was independent of sequence. The whole-body images clearly reflected the improved tumor to kidney ratios. By choosing a cytosine-free base sequence for pretargeting studies, kidney accumulation of cMORF-99mTc was reduced without adversely influencing tumor accumulation. The lowering of kidney radioactivity levels in this way may be important to reduce toxicity to this organ in connection with pretargeting radiotherapy studies.
KeywordsPretargeting Morpholino oligomer Tumor 99mTc Tumor imaging
- 1.Goodwin DA, Meares CF, McTigue M, et al. Rapid localization of haptens in sites containing previously administered antibody for immunoscintigraphy with short half-life tracers [abstract]. J Nucl Med. 1986; 27:959.Google Scholar
- 11.Wang Y, Chang F, Zhang Y, Liu N, Liu G, Gupta S, Rusckowski M, Hnatowich DJ. Pretargeting with amplification using polymeric peptide nucleic acid. Bioconjug Chem. 2001; 12:807–816.Google Scholar
- 12.He J, Liu G, Zhang S, Vanderheyden J-L, Liu N, Liu C, Zhang Y, Gupta S, Rusckowski M, Hnatowich DJ. A comparison of in vitro and in vivo stability in mice of two morpholino duplexes differing in chain length. Bioconjug Chem 2003, 14:1018–1023.Google Scholar
- 20.Rustom R, Maltby P, Grime JS, Stockdale HR, Critchley M, Bone JM. Effects of lysine infusion on the renal metabolism of aprotinin (Trasylol) in man. Clin Sci (Lond) 1992; 83:295–299.Google Scholar