The Impact Of The Microenvironment On Heterogeneity And Trametinib Response In Hcc1143 Triple Negative Breast Cancer Cells

Triple negative breast cancer (TNBC) lacks expression of hormone receptors (ER and PR) and HER2 and is characterized by aggressive disease with poor outcomes. Recent work suggests that TNBC also has a high degree of intratumoral heterogeneity, as measured by lineage differentiation status. This heterogeneity may impact therapeutic response, as it has been shown that treatment with PI3K/mTOR (BEZ235) or MEK (trametinib) inhibitors can drive TNBC cells into more homogeneous states, but that the surviving cells are resistant to the targeted therapy. In this study, we sought to understand how the microenvironment impacts differentiation state heterogeneity and response to targeted therapeutics in HCC1143 cells using our microenvironment microarray (MEMA) platform. Under low serum growth conditions, we found that several ligands could drive the growth of HCC1143, particularly EGF family ligands like AREG and EGF. With respect to differentiation state and heterogeneity, EGF and TGFB1 drove HCC1143 cells into a more mesenchymal like state, with increased expression of VIM and decreased expression of KRT14. In contrast, BMP2 led to higher levels of KRT14 and lower levels of VIM, leading to a more basal-like state. We also grew HCC1143 on MEMA with trametinib treatment. Here we found that combinations of collagen-based substrates and NRG1, HGF, and EGF ligands all led to higher cell counts and EdU incorporation rates compared to PBS-control treated cells. However, the levels of resistance conferred by the microenvironment was less than we had previously seen in HER2 positive MEMA, as the GR50 values (dose required to inhibit growth by 50%) only increased modestly (18 nM for untreated cells, 40 nM for NRG1, 45 nM for HGF). Interestingly, in that HER2 positive MEMA study, we identified HGF and NRG1 as potent resistance factors to lapatinib, but that they functioned in a subtype specific manner. HGF was effective in basal subtype cells and NRG1 in luminal, but not vice versa. We postulated that the modest resistance we observed was due to ligands acting on subsets of cells. We thus treated cells with a combination of NRG1 plus HGF, and found that this resulted in increased resistance (GR50= 91 nM). Imaging showed that trametinib drove HCC1143 cells to a homogenous KRT14 positive state, but surprisingly, addition of ligands reverted the cells to a more heterogeneous state that was resistant to trametinib. These data demonstrate that the microenvironment can impact the differentiation state of TNBC cells and is also capable of conferring resistance within subsets of the heterogeneous cell populations. Citation Format: Rebecca Smith, Kaylyn Devlin, Moqing Liu, Tiera Liby, David Kilburn, Elmar Bucher, Damir Sudar, Guillaume Thibault, Mark Dane, Joe Gray, Laura Heiser, James E. Korkola. The impact of the microenvironment on heterogeneity and trametinib response in HCC1143 triple negative breast cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1870.