Fig. 6: HER2 Heterogeneity scores derived from clustering analysis reveal correlation to clinical outcome.

Following the selection of qualified ER, PR, HER2, AR, and p53 CyCIF antibodies, the expression of selected antibodies was evaluated at a single-cell level in 567 HER2+ invasive breast cancer samples, representing 189 patients. Tissues from HER2+ patients (n = 77) in which there were at least 500 cells pooled from the triplicate cores were used for ITH analysis. a HER2 expression was analyzed in single cells, and the coefficient of variation (C.V.) among patients was plotted (y axis) by recurrence status. b HER2 and Ki67 mean intensity expression measured by CyCIF. c Distribution of cells across all clusters (blue) and HER2 core number 113 (orange) and d representative tumor with low (HER2-5 and HER2-161) and high (HER2-164 and HER2-170) heterogeneity. e HER2 heterogeneity scores were generated by identifying cells from each tissue mapped to the entire t-SNE. A larger boundary corresponds with higher diversity. f Samples that have equal distribution of each cluster have high heterogeneity and are diamond-shaped in the boundary mapping. g GMM and t-SNE scores reveal an association with recurrence. h Patients treated with Trastuzumab were removed from the GMM and t-SNE score analysis.