Abstract P1-08-10: Introduction of H1047R oncogenic mutation of PI3K p110alpha subunit in HER2-overexpressing mammary epithelial cells confers a "stem-like" phenotype and acute sensitivity to HSP90 inhibition
Authors: Surendran S , Bhola N , Arteaga CL , Chakraborty K , Chakrabarty A 
Published: March 2017

The Human Epidermal Growth Factor Receptor 2 (HER2) oncogene is amplified in one-fifth of breast cancers (BC). However, development of resistance against standard anti-HER2 therapies poses a major clinical challenge. Anti-tumor efficacy of HER2-targeting agents depends on inhibition of the downstream phosphatidylinositol-3 kinase (PI3K) signaling cascade. Gain-of-function somatic mutations in the gene encoding the PI3K catalytic subunit p110alpha (PIK3CA), co-expressed in about 40% of HER2+ BC, have been implicated in conferring resistance to HER2 monoclonal antibody herceptin. The single amino acid alteration H1047R within the kinase domain of PIK3CA is one of three hot spot mutations prevalent in BC.

Previously, we demonstrated that introduction of H1047R mutation in HER2-overexpressing MCF10A mammary epithelial cells enhances cellular transformation and decreases herceptin sensitivity by inducing secretion of endogenous ErbB ligand heregulin. However, genetic ablation of HER3, the major co-receptor for HER2 and the solitary receptor for heregulin, was insufficient for complete inhibition of cell growth, indicating the existence of additional mechanism/s responsible for the heightened aggressiveness and decreased drug sensitivity of HER2/H1047RPI3K cells. In the current study, we looked further into the molecular changes within these cells that might be responsible for these phenomena.

When compared with the HER2/WTPI3K cells, the HER2/H1047RPI3K cells revealed a significant increase in CD44high/CD24low/negative populations, common markers of BC stem cells, as well as molecular and phenotypic changes associated with epithelial-to-mesenchymal transition. These observations are in agreement with previously published report on mouse model of HER2/H1047RPI3K BC. Further analyses demonstrated additional stem cell-associated characteristics in HER2/H1047RPI3K cells, such as expression of angiogenic and inflammatory cytokines, ability to induce chemotaxis and invasion, activation of TGFb and NFKb signaling pathways. Connectivity map (CMap) analysis of the gene expression signatures from HER2/H1047RPI3K cells revealed a negative association with those from BC cells treated with 17AAG, an inhibitor of the heat shock protein 90 (HSP90). In line with this, HER2/H1047RPI3K-expressing cells are found to be more sensitive to HSP90 inhibition compared to the pan-ErbB inhibitor lapatinib.

Cancer stem cells are implicated in drug resistance and tumor recurrence. Enrichment of cell population expressing high levels of stem cell markers and stem cell-related features could be one of major mechanisms by which BC cells co-expressing HER2 and H1047RPI3K adapt to anti-HER2 therapeutic agents. Acute dependence on molecular chaperone HSP90 provides a unique, yet practical opportunity to effectively inhibit tumors harboring both molecular alterations, since HSP90 inhibitors have already shown encouraging clinical activity in herceptin-resistant setting.

Citation Format: Surendran S, Bhola N, Arteaga CL, Chakraborty K, Chakrabarty A. Introduction of H1047R oncogenic mutation of PI3K p110alpha subunit in HER2-overexpressing mammary epithelial cells confers a "stem-like" phenotype and acute sensitivity to HSP90 inhibition [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-08-10.