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Heterogeneous HER2 antigen expression among individual Circulating Tumor Cells within breast cancer patients Julie Smith, DVM, PHD Postdoc / lecturer @ Biomedical Laboratory Science, Department of Technology DET SUNDHEDSFAGLIGE OG TEKNOLOGISKE FAKULTET Just so you know…. Agenda! Background: - HER2 expression in relation to breast cancer? - Primary tumors vs metastases – How do we treat? - What are CTCs? A liquid Biopsy? CytoTrack Materials and Methods: - Development of HER2 characterization on Tumor Cell Lines - Testing HER2 characterisation on Breast Cancer Patients Results: - The HER2 ICC method - HER2 on CTCs of 7 breast cancer patients Conclusion and round off www.phmetropol.dk BACKGROUND www.phmetropol.dk BACKGROUND HER2 neg HER2 pos www.phmetropol.dk BACKGROUND ? www.phmetropol.dk CTC = circulating tumour cells EMT = epithelial-to-mesenchymal transition MET = mesenchymal-to-epithelial transition DTC = disseminated tumour cells BACKGROUND Liquid Biopsy? Scanning Imaging HER2 ICC/FISH ICC = ImmuneCytoChemistry staining Revisting Reimaging www.phmetropol.dk MATERIALS AND METHODS To develop: HER2-staining protocol for CytoDisc: Tumor Cell Lines spiked in healthy donor blood Mix of known HER2 negative and positive (SKBR-3, CALU-3, MB231, MCF7) To test: the developed HER2 staining protocol in vivo Breast Cancer Patients, n = 11 from The Maria Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Warsaw, Poland. www.phmetropol.dk RESULTS Tumor cell line studies: Incubate CytoDisc with CTCs in: • 1:50 Anti-HER2 diluted in PBS buffer • For 1 hour • In darkness • At room temperature www.phmetropol.dk RESULTS Breast cancer in vivo studies: CTCs were detected in 7 of 11 patients ranging from 1 to 189 CTC. Table 1. CTC Enumeration and Characterization Patient CTC total (n) HER2+ (n) HER2+ (%) A 189 0 0% B 1 0 0% C 0 0 - D 5 0 0% E 0 0 - F 0 0 - G 14 12 86% H 2 2 100% I 22 7 32% J 4 4 100% K 0 0 - www.phmetropol.dk CONCLUSION A novel method for CTC enumeration and HER2 characterization is described. CTC heterogeneity was observed in breast cancer patients. Future clinical studies are on-going to examine the clinical significance of these findings. www.phmetropol.dk THANK YOU www.phmetropol.dk ROUND OFF www.phmetropol.dk