Abstract
Purpose
Overexpression of HER2/neu in breast cancer is correlated with a poor prognosis. It may vary between primary tumors and metastatic lesions and change during the treatment. Therefore, there is a need for a new means to assess HER2/neu expression in vivo. In this work, we used 68Ga-labeled DOTA-ZHER2:2891-Affibody to monitor HER2/neu expression in a panel of breast cancer xenografts.
Methods
DOTA-ZHER2:2891-Affibody molecules were labeled with 68Ga. In vitro binding was characterized by a receptor saturation assay. Biodistribution and PET imaging studies were conducted in athymic nude mice bearing subcutaneous human breast cancer tumors with three different levels of HER2/neu expression. Nonspecific uptake was analyzed using non-HER2-specific Affibody molecules. Signal detected by PET was compared with ex vivo assessment of the tracer uptake and HER2/neu expression.
Results
The 68Ga-DOTA-ZHER2:2891-Affibody probe showed high binding affinity to MDA-MB-361 cells (K D = 1.4 ± 0.19 nM). In vivo biodistribution and PET imaging studies demonstrated high radioactivity uptake in HER2/neu-positive tumors. Tracer was eliminated quickly from the blood and normal tissues, resulting in high tumor-to-blood ratios. The highest concentration of radioactivity in normal tissue was seen in the kidneys (227 ± 14%ID/g). High-contrast PET images of HER2/neu-overexpressing tumors were recorded as soon as 1 h after tracer injection. A good correlation was observed between PET imaging, biodistribution estimates of tumor tracer concentration, and the receptor expression.
Conclusion
These results suggest that PET imaging using 68Ga-DOTA-ZHER2:2891-Affibody is sensitive enough to detect different levels of HER2/neu expression in vivo.
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References
Tagliabue E, Balsari A, Campiglio M, Pupa SM. HER2 as a target for breast cancer therapy. Expert Opin Biol Ther. 2010;10:711–24.
Dean-Colomb W, Esteva FJ. Her2-positive breast cancer: herceptin and beyond. Eur J Cancer. 2008;44:2806–12.
Pohlmann PR, Mayer IA, Mernaugh R. Resistance to trastuzumab in breast cancer. Clin Cancer Res. 2009;15:7479–91.
Allison M. The HER2 testing conundrum. Nat Biotechnol. 2010;28:117–9.
Dowsett M, Hanna WM, Kockx M, Penault-Llorca F, Ruschoff J, Gutjahr T, et al. Standardization of HER2 testing: results of an international proficiency-testing ring study. Mod Pathol. 2007;20:584–91.
Capala J, Bouchelouche K. Molecular imaging of HER2-positive breast cancer: a step toward an individualized 'image and treat' strategy. Curr Opin Oncol. 2010;22:559–66.
Wester HJ. Nuclear imaging probes: from bench to bedside. Clin Cancer Res. 2007;13:3470–81.
Fass L. Imaging and cancer: a review. Mol Oncol. 2008;2:115–52.
Fani M, Andre JP, Maecke HR. 68Ga-PET: a powerful generator-based alternative to cyclotron-based PET radiopharmaceuticals. Contrast Media Mol Imaging. 2008;3:67–77.
Maecke HR, Hofmann M, Haberkorn U. 68Ga-labeled peptides in tumor imaging. J Nucl Med. 2005;46:172S–8.
Al-Nahhas A, Win Z, Szyszko T, Singh A, Khan S, Rubello D. What can gallium-68 PET add to receptor and molecular imaging? Eur J Nucl Med Mol Imaging. 2007;34:1897–901.
Partridge M, Spinelli A, Ryder W, Hindorf C. The effect of beta(+) energy on performance of a small animal PET camera. Nucl Instrum Methods Phys Res A. 2006;568:933–6.
Baum RP, Prasad V, Muller D, Schuchardt C, Orlova A, Wennborg A, et al. Molecular imaging of HER2-expressing malignant tumors in breast cancer patients using synthetic 111In- or 68Ga-labeled affibody molecules. J Nucl Med. 2010;51:892–7.
Lofblom J, Feldwisch J, Tolmachev V, Carlsson J, Stahl S, Frejd FY. Affibody molecules: engineered proteins for therapeutic, diagnostic and biotechnological applications. FEBS Lett. 2010;584:2670–80.
Kramer-Marek G, Kiesewetter DO, Martiniova L, Jagoda E, Lee SB, Capala J. [18F]FBEM-Z(HER2:342)-Affibody molecule – a new molecular tracer for in vivo monitoring of HER2 expression by positron emission tomography. Eur J Nucl Med Mol Imaging. 2008;35:1008–18.
Tolmachev V, Velikyan I, Sandstrom M, Orlova A. A HER2-binding Affibody molecule labelled with 68Ga for PET imaging: direct in vivo comparison with the 111In-labelled analogue. Eur J Nucl Med Mol Imaging. 2010;37:1356–67.
Kramer-Marek G, Kiesewetter DO, Capala J. Changes in HER2 expression in breast cancer xenografts after therapy can be quantified using PET and (18)F-labeled affibody molecules. J Nucl Med. 2009;50:1131–9.
Ahlgren S, Orlova A, Rosik D, Sandstrom M, Sjoberg A, Baastrup B, et al. Evaluation of maleimide derivative of DOTA for site-specific labeling of recombinant affibody molecules. Bioconjug Chem. 2008;19:235–43.
Blasberg RG. Molecular imaging and cancer. Mol Cancer Ther. 2003;2:335–43.
Tolmachev V. Imaging of HER-2 overexpression in tumors for guiding therapy. Curr Pharm Des. 2008;14:2999–3019.
Zidan J, Dashkovsky I, Stayerman C, Basher W, Cozacov C, Hadary A. Comparison of HER-2 overexpression in primary breast cancer and metastatic sites and its effect on biological targeting therapy of metastatic disease. Br J Cancer. 2005;93:552–6.
McLarty K, Reilly RM. Molecular imaging as a tool for personalized and targeted anticancer therapy. Clin Pharmacol Ther. 2007;81:420–4.
Feldwisch J, Tolmachev V, Lendel C, Herne N, Sjoberg A, Larsson B, et al. Design of an optimized scaffold for affibody molecules. J Mol Biol. 2010;398:232–47.
Miao Z, Levi J, Cheng Z. Protein scaffold-based molecular probes for cancer molecular imaging. Amino Acids 2010. doi:10.1007/s00726-010-0503-9
Eigenbrot C, Ultsch M, Dubnovitsky A, Abrahmsen L, Hard T. Structural basis for high-affinity HER2 receptor binding by an engineered protein. Proc Natl Acad Sci U S A. 2010;107:15039–44.
Ahlgren S, Tolmachev V. Radionuclide molecular imaging using Affibody molecules. Curr Pharm Biotechnol. 2010;11:581–9.
De Lorenzo C, Cozzolino R, Carpentieri A, Pucci P, Laccetti P, D'Alessio G. Biological properties of a human compact anti-ErbB2 antibody. Carcinogenesis. 2005;26:1890–5.
Perk LR, Visser GW, Vosjan MJ, Stigter-van Walsum M, Tijink BM, Leemans CR, et al. (89)Zr as a PET surrogate radioisotope for scouting biodistribution of the therapeutic radiometals (90)Y and (177)Lu in tumor-bearing nude mice after coupling to the internalizing antibody cetuximab. J Nucl Med. 2005;46:1898–906.
Smith-Jones PM, Solit DB, Akhurst T, Afroze F, Rosen N, Larson SM. Imaging the pharmacodynamics of HER2 degradation in response to Hsp90 inhibitors. Nat Biotechnol. 2004;22:701–6.
Ren G, Zhang R, Liu Z, Webster JM, Miao Z, Gambhir SS, et al. A 2-helix small protein labeled with 68Ga for PET imaging of HER2 expression. J Nucl Med. 2009;50:1492–9.
Orlova A, Wallberg H, Stone-Elander S, Tolmachev V. On the selection of a tracer for PET imaging of HER2-expressing tumors: direct comparison of a 124I-labeled affibody molecule and trastuzumab in a murine xenograft model. J Nucl Med. 2009;50:417–25.
McLarty K, Cornelissen B, Scollard DA, Done SJ, Chun K, Reilly RM. Associations between the uptake of 111In-DTPA-trastuzumab, HER2 density and response to trastuzumab (Herceptin) in athymic mice bearing subcutaneous human tumour xenografts. Eur J Nucl Med Mol Imaging. 2009;36:81–93.
van Dalen J, Visser E, Laverman P, Vogel W, Oyen W, Corstens F, Boerman O. Effect of the positron range on the spatial resolution of a new generation pre-clinical PET-scanner using F-18, Ga-68, Zr-89 and I-124. J Nucl Med. 2008;49 Suppl 1:404P
Yang DJ, Azhdarinia A, Kim EE. Tumor specific imaging using Tc-99m and Ga-68 labeled radiopharmaceuticals. Curr Med Imaging Rev. 2005;1:25–34.
Aerts HJ, Dubois L, Perk L, Vermaelen P, van Dongen GA, Wouters BG, et al. Disparity between in vivo EGFR expression and 89Zr-labeled cetuximab uptake assessed with PET. J Nucl Med. 2009;50:123–31.
Tolmachev V, Nilsson FY, Widstrom C, Andersson K, Rosik D, Gedda L, et al. 111In-benzyl-DTPA-ZHER2:342, an affibody-based conjugate for in vivo imaging of HER2 expression in malignant tumors. J Nucl Med. 2006;47:846–53.
Rolleman EJ, Bernard BF, Breeman WA, Forrer F, de Blois E, Hoppin J, et al. Molecular imaging of reduced renal uptake of radiolabelled [DOTA0,Tyr3]octreotate by the combination of lysine and Gelofusine in rats. Nuklearmedizin. 2008;47:110–5.
Acknowledgments
The authors appreciate the support of Affibody AB. We owe special thanks to Michael Green for constructive discussions. We also appreciate the technical assistance of Ilya Lyakhov, Monika Kuban, and Alesia Holly.
The contents of this article do not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organization imply endorsement by the U.S. Government.
This research was supported in part by the Center for Cancer Research, an Intramural Research Program of the National Cancer Institute, the Imaging Probe Development Center, the National Heart, Lung, and Blood Institute, and the Breast Cancer Research Stamp Fund (awarded through competitive peer review), and was funded in part with Federal funds from the National Cancer Institute, National Institutes of Health, under contracts N01-CO-12400 and N01-CO-12401.
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Kramer-Marek, G., Shenoy, N., Seidel, J. et al. 68Ga-DOTA-Affibody molecule for in vivo assessment of HER2/neu expression with PET. Eur J Nucl Med Mol Imaging 38, 1967–1976 (2011). https://doi.org/10.1007/s00259-011-1810-4
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DOI: https://doi.org/10.1007/s00259-011-1810-4