Multiplexed Measurement of Signaling Targets and Cytotoxic T cells in Trastuzumab-treated Patients using Imaging Mass Cytometry

Purpose: Imaging Mass Cytometry (IMC) uses metal-conjugated antibodies to provide multidimensional, objective measurement of protein targets. We used this high-throughput platform to perform an 18-plex assessment of HER2 ICD/ECD, cytotoxic T cell infiltration and other structural and signaling proteins in a cohort of patients treated with trastuzumab to discover associations with trastuzumab benefit. Experimental Design: An antibody panel for detection of 18 targets (Pancytokeratin, HER2 ICD, HER2 ECD, CD8, vimentin, cytokeratin 7, beta-catenin, HER3, MET, EGFR, ERK 1-2, MEK 1-2, PTEN, PI3K p110 alpha, Akt, mTOR, Ki67 and Histone H3) was used with a selection of trastuzumab-treated patients from the HeCOG 10/05 trial (n=180), and identified a case:control series. Results: Patients that recurred after adjuvant treatment with trastuzumab trended toward a decreased fraction of HER2 ECD pixels over threshold compared to cases without recurrence (p=0.057). After exclusion of the lowest HER2 expressers, 5-year recurrence events where associated with reduced total ECD/ ICD ratio intensity in tumor (p=0.044). These observations are consistent with our previous work using QIF but represents the proof on identical cell content. We also describe the association of the extracellular domain of HER2 with CD8 T-cell infiltration on the same slide. Conclusions: The proximity of CD8 cells as a function of the expression of the ECD of HER2 provides further evidence for the role of the immune system in the mechanism of action of trastuzumab.

https://ift.tt/2U9DSUZ