Abstract
Molecular imaging of human epidermal growth factor receptor type 2 (HER2) expression has drawn significant attention because of the unique role of the HER2 gene in diagnosis, therapy and prognosis of human breast cancer. In our previous research, a novel cyclic 2-helix small protein, MUT-DS, was discovered as an anti-HER2 Affibody analog with high affinity through rational protein design and engineering. MUT-DS was then evaluated for positron emission tomography (PET) of HER2-positive tumor by labeling with two radionuclides, 68Ga and 18F, with relatively short half-life (t 1/2 < 2 h). In order to fully study the in vivo behavior of 2-helix small protein and demonstrate that it could be a robust platform for labeling with a variety of radionuclides for different applications, in this study, MUT-DS was further radiolabeled with 64Cu or 111In and evaluated for in vivo targeting of HER2-positive tumor in mice. Design 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) conjugated MUT-DS (DOTA–MUT-DS) was chemically synthesized using solid phase peptide synthesizer and I2 oxidation. DOTA–MUT-DS was then radiolabeled with 64Cu or 111In to prepare the HER2 imaging probe (64Cu/111In-DOTA–MUT-DS). Both biodistribution and microPET imaging of the probe were evaluated in nude mice bearing subcutaneous HER2-positive SKOV3 tumors. DOTA–MUT-DS could be successfully synthesized and radiolabeled with 64Cu or 111In. Biodistribution study showed that tumor uptake value of 64Cu or 111In-labeled DOTA–MUT-DS was 4.66 ± 0.38 or 2.17 ± 0.15%ID/g, respectively, in nude mice bearing SKOV3 xenografts (n = 3) at 1 h post-injection (p.i.). Tumor-to-blood and tumor-to-muscle ratios for 64Cu-DOTA-MUT-DS were attained to be 3.05 and 3.48 at 1 h p.i., respectively, while for 111In-DOTA–MUT-DS, they were 2.04 and 3.19, respectively. Co-injection of the cold Affibody molecule ZHER2:342 with 64Cu-DOTA-MUT-DS specifically reduced the SKOV3 tumor uptake of the probe by 48%. 111In-DOTA–MUT-DS displayed lower liver uptake at all the time points investigated and higher tumor to blood ratios at 4 and 20 h p.i., when compared with 64Cu-DOTA–MUT-DS. This study demonstrates that the 2-helix protein based probes, 64Cu/111In DOTA–MUT-DS, are promising molecular probes for imaging HER2-positive tumor. Two-helix small protein scaffold holds great promise as a novel and robust platform for imaging and therapy applications.
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Abbreviations
- DOTA:
-
1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid
- PET:
-
Positron emission tomography
- SPECT:
-
Single photon emission computed tomography
- HPLC:
-
High-performance liquid chromatography
- p.i.:
-
Postinjection
- MW:
-
Molecular weight
- OSEM:
-
Ordered subsets expectation maximum
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Acknowledgments
This work was supported, in part, by California Breast Cancer Research Program 14IB-0091 (ZC) and SNM Pilot Research Grant (ZC), Medical Diagnostics, GE Healthcare, and National Cancer Institute (NCI) Small Animal Imaging Resource Program (SAIRP) grant R24 CA93862. We also thank Dr. Joshua Hoerner, Gregory Goddard and Hans Grade of GE Global Research for MS analysis and Dr. Alex Gibson of GE Healthcare for their helpful suggestion and reviewing of the manuscript.
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Ren, G., Webster, J.M., Liu, Z. et al. In vivo targeting of HER2-positive tumor using 2-helix affibody molecules. Amino Acids 43, 405–413 (2012). https://doi.org/10.1007/s00726-011-1096-7
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DOI: https://doi.org/10.1007/s00726-011-1096-7