Comparison of tumor‑targeting properties of directly and indirectly radioiodinated designed ankyrin repeat protein (DARPin) G3 variants for molecular imaging of HER2

Vorobyeva,Anzhelika; Sсhulga,Alexey; Konovalova,Elena; Güler,Rezan; Mitran,Bogdan; Garousi,Javad; Rinne,Sara; Löfblom,John; Orlova,Anna; Deyev,Sergey; Tolmachev,Vladimir

doi:10.3892/ijo.2019.4712

Comparison of tumor‑targeting properties of directly and indirectly radioiodinated designed ankyrin repeat protein (DARPin) G3 variants for molecular imaging of HER2

Authors:
- Anzhelika Vorobyeva
- Alexey Sсhulga
- Elena Konovalova
- Rezan Güler
- Bogdan Mitran
- Javad Garousi
- Sara Rinne
- John Löfblom
- Anna Orlova
- Sergey Deyev
- Vladimir Tolmachev
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Affiliations: Department of Immunology, Genetics and Pathology, Uppsala University, SE 75185 Uppsala, Sweden, Molecular Immunology Laboratory, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia, Department of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, SE 10691 Stockholm, Sweden, Department of Medicinal Chemistry, Uppsala University, SE 75183 Uppsala, Sweden
Published online on: February 11, 2019 https://doi.org/10.3892/ijo.2019.4712
Pages: 1209-1220
Copyright: © Vorobyeva et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Evaluation of human epidermal growth factor receptor 2 (HER2) expression levels in breast and gastroesophageal cancer is used for the stratification of patients for HER2‑targeting therapies. The use of radionuclide molecular imaging may facilitate such evaluation in a non‑invasive way. Designed ankyrin repeat proteins (DARPins) are engineered scaffold proteins with high potential as probes for radionuclide molecular imaging. DARPin G3 binds with high affinity to HER2 and may be used to visualize this important therapeutic target. Studies on other engineered scaffold proteins have demonstrated that selection of the optimal labeling approach improves the sensitivity and specificity of radionuclide imaging. The present study compared two methods of labeling G3, direct and indirect radioiodination, to select an approach providing the best imaging contrast. G3‑H6 was labeled with iodine‑124, iodine‑125 and iodine‑131 using a direct method. A novel construct bearing a C‑terminal cysteine, G3‑GGGC, was site‑specifically labeled using [125I]I‑iodo‑[(4‑hydroxyphenyl)ethyl]maleimide (HPEM). The two radiolabeled G3 variants preserved binding specificity and high affinity to HER2‑expressing cells. The specificity of tumor targeting in vivo was demonstrated. Biodistribution comparison of [131I]I‑G3‑H6 and [125I]I‑HPEM‑G3‑GGGC in mice, bearing HER2‑expressing SKOV3 xenografts, demonstrated an appreciable contribution of hepatobiliary excretion to the clearance of [125I]I‑HPEM‑G3‑GGGC and a decreased tumor uptake compared to [131I]I‑G3‑H6. The direct label provided higher tumor‑to‑blood and tumor‑to‑organ ratios compared with the indirect label at 4 h post‑injection. The feasibility of high contrast PET/CT imaging of HER2 expression in SKOV3 xenografts in mice using [124I]I‑G3‑H6 was demonstrated. In conclusion, direct radioiodination is the preferable approach for labeling DARPin G3 with iodine‑123 and iodine‑124 for clinical single photon emission computed tomography and positron emission tomography imaging.

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