Comparative evaluation of 111In-labeled NOTA‑conjugated affibody molecules for visualization of HER3 expression in malignant tumors

Andersson,Ken  G.; Rosestedt,Maria; Varasteh,Zohreh; Malm,Magdalena; Sandström,Mattias; Tolmachev,Vladimir; Löfblom,John; Ståhl,Stefan; Orlova,Anna

doi:10.3892/or.2015.4046

Comparative evaluation of 111In-labeled NOTA‑conjugated affibody molecules for visualization of HER3 expression in malignant tumors

Authors:
- Ken G. Andersson
- Maria Rosestedt
- Zohreh Varasteh
- Magdalena Malm
- Mattias Sandström
- Vladimir Tolmachev
- John Löfblom
- Stefan Ståhl
- Anna Orlova
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Affiliations: Division of Protein Technology, KTH Royal Institute of Technology, Stockholm, Sweden, Preclinical PET Platform, Uppsala University, Uppsala, Sweden, Nuclear Medicine and PET, Uppsala University, Uppsala, Sweden, Institute of Immunology, Genetic and Pathology, Uppsala University, Uppsala, Sweden
Published online on: June 9, 2015 https://doi.org/10.3892/or.2015.4046
Pages: 1042-1048

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Abstract

Expression of human epidermal growth factor receptor type 3 (HER3) in malignant tumors has been associated with resistance to a variety of anticancer therapies. Several anti-HER3 monoclonal antibodies are currently under pre-clinical and clinical development aiming to overcome HER3-mediated resistance. Radionuclide molecular imaging of HER3 expression may improve treatment by allowing the selection of suitable patients for HER3-targeted therapy. Affibody molecules are a class of small (7 kDa) high-affinity targeting proteins with appreciable potential as molecular imaging probes. In a recent study, we selected affibody molecules with affinity to HER3 at a low picomolar range. The aim of the present study was to develop an anti-HER3 affibody molecule suitable for labeling with radiometals. The HEHEHE-Z08698-NOTA and HEHEHE-Z08699-NOTA HER3-specific affibody molecules were labeled with indium‑111 (111In) and assessed in vitro and in vivo for imaging properties using single photon emission computed tomography (SPECT). Labeling of HEHEHE-Z08698-NOTA and HEHEHE-Z08699-NOTA with 111In provided stable conjugates. In vitro cell tests demonstrated specific binding of the two conjugates to HER3-expressing BT‑474 breast carcinoma cells. In mice bearing BT‑474 xenografts, the tumor uptake of the two conjugates was receptor‑specific. Direct in vivo comparison of 111In-HEHEHE-Z08698-NOTA and 111In-HEHEHE-Z08699‑NOTA demonstrated that the two conjugates provided equal radioactivity uptake in tumors, although the tumor-to-blood ratio was improved for 111In-HEHEHE-Z08698-NOTA [12±3 vs. 8±1, 4 h post injection (p.i.)] due to more efficient blood clearance. 111In-HEHEHE-Z08698-NOTA is a promising candidate for imaging of HER3-expression in malignant tumors using SPECT. Results of the present study indicate that this conjugate could be used for patient stratification for anti-HER3 therapy.

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