⁶⁸Ga-, ⁸⁶Y- and ¹¹¹In-radiolabeled anti-tenascin-C oligonucleotide aptamers as a potential probe for tumor imaging

Introduction:

The matrix protein tenascin-c (TN-C) represents an interesting target for molecular imaging in oncology due to it´s high abundance in a variety of human tumors such as lung, breast and brain tumors [1]. First radiolabeled anti-TN-C antibodies could successfully proof the concept of tumor-TN-C targeting [2]. However, a persistently high blood level hampers their use for targeted imaging. Aptamers, a class of rapidly clearing high affinity oligonucleotides, have been introduced successfully to molecular imaging by the application of the Tc-99m labeled TN-C Targeting Aptamer-1 (TTA-1) [3]. Here we report on the radiolabeling, binding affinity and biodistribution of this novel targeting aptamer labeled with metal PET isotopes.

Experimental:

Labeling with Ga(III), Y(III), and In(III) was performed in a one pot reaction after conjugation of a DOTA-type chelating moiety to TTA-1-MAG2. In-111 was included in the study as a surrogate for Ga and Y in long term stability and affinity tests. The labeling was carried out in the presence of the aptamer-chelator conjugate and the respective radiometal in acetate buffer. Degradation was investigated in human plasma employing PAGE. Affinity for human TN-C was determined in a nitrous cellulose filter binding assay. Athymic mice, bearing the human U251 tumor cell line, were injected with the radiolabeled TTA-1 derivatives.

Results and Discussion:

Addition of the radiometal salts yielded the respective labeled aptamer conjugates in high yields (60 – 80 %). Ultrafiltration of the products led to > 95 % of pure material as determined by PAGE and HPLC methods. The biological stability in human plasma ranged from 68 % (In-111) to 95 % (Y-86) of intact material after 6 hr. Binding affinity for human TN-C revealed a KD of 1 nM (In-111). A maximum tumor uptake of 2.0 % ID/g (Ga-68), 1 hr p.i. and T/Non Tumor ratios of 5.1 (T/blood), 0.2 (T/kidney), 0.7 (T/liver) 1 hr p.i. proved to be adequate to image the U251 tumor xenograft in mice. Tumor visualization was possible for both Ga-68 and Y-86 labeled TTA-1.

Conclusion:

The introduction of a DOTA-type chelating moiety could be successfully used to radiolabel the aptamer probe with In, Ga and Y. The stability against nuclease degradation along with a significant tumor accumulation, high T/ blood levels and the imaging capabilities in PET-scans make these compounds promising candidates for further evaluation as multi-tumor imaging agents.

Acknowledgement:

1. R. Chiquet-Ehrismann, Cell 47, 131 (1986)
2. P. Riva, Cancer 73, 1076 (1994)
3. B.J. Hicke, J. Nuc. Med. 47, 668-678 (2006).