Abstract S4-02: Integrated genomic analyses of members of protein kinase C family identifies subtype specific alterations as novel therapeutic targets

Abstract

Abstract Introduction Members of the protein kinase C family are serine/threonine kinases that are involved in proliferation, apoptosis, cell survival and migration, and have been implicated in tumorigenesis. Recently PRKCE has been found to be up-regulated in triple-negative (TN) breast cancers and has been proposed as a target for therapeutic intervention. The aims of this study were to determine i) whether different members of the PKC family are dysregulated and/ or mutated in specific subtypes of breast cancer, ii) to investigate the impact of silencing or overexpression of members of the PKC family in cell line models representative of the different breast cancer subtypes. Material and methods: We obtained expression and mutational data from the cancer genome atlas (TCGA) project from 567 and 640 samples subjected to microarray-based gene expression profiling and whole exome sequencing, respectively. Pair-wise SAM analysis of TCGA gene expression data was performed to identify differential expression between subgroups (ER+, HER2+ and TN). Potential driver mutations were identified through the algorithm CHASM. Subtype specific dependencies were identified from the re-analysis of publicly available siRNA kinome-wide screens in a panel of 20 breast cancer cell lines. In vitro assessment of gene overexpression was assessed in MCF10A cells by wound healing scratch assays and 3D growth in Matrigel. Results PRKCA, B, I and Q were expressed at significantly higher levels in TN breast cancers. Higher expression of PRKCE was significantly associated with ER-negativity, whereas high PRKCD expression was associated with ER-positivity. Analysis of siRNA kinome-wide screen data resulted in a significant reduction in survival associated with PRKCI and PRKCE in ER-negative cells, PRKCQ in triple-negative cells, and PRKCD in ER-positive cells. Furthermore meta-analysis of published exome and whole genome sequencing data identified potentially activating recurrent kinase domain mutations in PRKCB (0.93%), Q (1.25%) and Z (0.93%), with mutations in PRKCZ being associated with higher gene expression. Forced expression of wild-type PRKCQ and PRKCZ in MCF10A cells resulted in the formation of irregular acini in 3D cell culture and expression of wild-type PRKCZ resulted in increased migration. Conclusions Differential expression of members of the protein kinase C family, are associated with different molecular subtypes of breast cancer. Furthermore, we have shown that breast cancer cells are dependent upon expression of these family members in vitro, which are associated with different cellular phenotypes. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr S4-02.