Measuring HER2-Receptor Expression In Metastatic Breast Cancer Using [68Ga]ABY-025 Affibody PET/CT

Jens Sörensen, Irina Velikyan, Dan Sandberg, Anders Wennborg, Joachim Feldwisch, Vladimir Tolmachev, Anna Orlova, Mattias Sandström, Mark Lubberink, Helena Olofsson, Jörgen Carlsson, Henrik Lindman, Jens Sörensen, Irina Velikyan, Dan Sandberg, Anders Wennborg, Joachim Feldwisch, Vladimir Tolmachev, Anna Orlova, Mattias Sandström, Mark Lubberink, Helena Olofsson, Jörgen Carlsson, Henrik Lindman

Abstract

Purpose: Positron Emission Tomography (PET) imaging of HER2 expression could potentially be used to select patients for HER2-targed therapy, predict response based on uptake and be used for monitoring. In this phase I/II study the HER2-binding Affibody molecule ABY-025 was labeled with (68)Ga-gallium ([(68)Ga]ABY-025) for PET to study effect of peptide mass, test-retest variability and correlation of quantified uptake in tumors to histopathology.

Experimental design: Sixteen women with known metastatic breast cancer and on-going treatment were included and underwent FDG PET/CT to identify viable metastases. After iv injection of 212±46 MBq [(68)Ga]ABY-025 whole-body PET was performed at 1, 2 and 4 h. In the first 10 patients (6 with HER2-positive and 4 with HER2-negative primary tumors), [(68)Ga]ABY-025 PET/CT with two different doses of injected peptide was performed one week apart. In the last six patients (5 HER2-positive and 1 HER2-negative primary tumors), repeated [(68)Ga]ABY-025 PET were performed one week apart as a test-retest of uptake in individual lesions. Biopsies from 16 metastases in 12 patients were collected for verification of HER2 expression by immunohistochemistry and in-situ hybridization.

Results: Imaging 4h after injection with high peptide content discriminated HER2-positive metastases best (p<0.01). PET SUV correlated with biopsy HER2-scores (r=0.91, p<0.001). Uptake was five times higher in HER2-positive than in HER2-negative lesions with no overlap (p=0.005). The test-retest intra-class correlation was r=0.996. [(68)Ga]ABY-025 PET correctly identified conversion and mixed expression of HER2 and targeted treatment was changed in 3 of the 16 patients.

Conclusion: [(68)Ga]ABY-025 PET accurately quantifies whole-body HER2-receptor status in metastatic breast cancer.

Keywords: 68Ga-gallium; Affibody; HER2-receptor; breast cancer metastases; clinical study.

Conflict of interest statement

Competing Interests: Anders Wennborg and Joachim Feldwisch are employees of Affibody AB.

Figures

Figure 1
Figure 1
Uptake images of [68Ga]ABY-025 with low (LD) and high administered peptide dose (HD) at 1, 2 and 4 hours after injection in patient 2. All images are normalized to the same scale of SUV 10. A rainbow color scale is used, red color: SUV>10. Normal soft tissue uptake was higher with LD. Note the gradually higher contrast of metastases in liver and bone (arrows), due to both disappearance of normal liver uptake and tumor uptake accumulation.
Figure 2
Figure 2
Maximum Intensity Projection PET images from two studied patients with wide-spread metastatic breast cancer. A: Patient 5 HER2-negative. B: Patient 7 HER2-positive. FDG-PET images (A1, B1) and [68Ga]ABY-025 PET images (A2=HER2-negative, B2=HER2-positive) are shown. All images are normalized to SUV 10. Darker colors indicate higher uptake.
Figure 3
Figure 3
Relation of PET Standard Uptake Values (SUVmax) of [68Ga]ABY-025 and corresponding HER2-scoring from combined immunohistochemistry and in situ hybridization in 16 biopsied metastases. PET images were obtained at 2 (all 16 patients) and 4 hours (10 patients) after injection of 212±46 MBq [68Ga]ABY-025 with peptide mass 427±19 μg (HD). A: correlation of SUVmax and HER2-score in 16 biopsied metastases at 2h imaging. A threshold of SUVmax=6 separated HER-negative from HER2-positive with an accuracy of 88%. Hollow squares indicate metastases from patients only scanned at 2h. B: correlation of SUVmax and HER2-score in 12 biopsied metastases at 4h imaging. At this time point separation of HER-negative from HER2-positive increased and a threshold of SUVmax=8 had an accuracy of 100%. C: The plot shows the range of [68Ga]ABY-025 SUVmax in all nodular metastases (black dots) identified using FDG-PET/CT in all 16 patients undergoing [68Ga]ABY-025 PET/CT at 2h after injection of tracer with high peptide content. Crosses symbolize biopsied metastases. The red line indicates a proposed threshold at SUVmax=6 for discriminating HER2-positive and HER2-negative metastases. Notheworthy, based on PET the majority of patients had heterogeneous HER2 expression with both HER2-positive and HER2-negative metastases.
Figure 4
Figure 4
Based on the results from [68Ga]ABY-025 PET/CT, mixed expression of HER2 in metastatic breast cancer was seen in several patients and confirmed by biopsies in 2. Patient 9 had HER2-negative primary tumor and was enrolled as negative control. FDG-PET/CT showed metastases in left liver lobe, peritoneal lymph nodes and cervix of uterus. ABY-025 uptake was high in the liver metastasis, low in peritoneal metastases and absent in the cervical region (not shown). According to IHC, the liver finding was true positive and both other sites were true negative.

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Source: PubMed

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