Bisphenol A Interacts with GPER, Activates EGFR and ERK Signaling and Antagonizes Efficacy of Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors in Breast Cancer Cells (Poster # SUN-0345) Scott J. Sauer, PhD, H. Kim Lyerly, MD, Imran Shah, PhD, Kevin P. Williams, PhD, Gayathri R. Devi, PhD Unmet Challenge in Inflammatory Breast Cancer (IBC) • One of the deadliest subtypes of locally advanced breast cancer – Rapid growth – Highly metastatic – Rare (1-5% of US breast cancers) but likely underreported Erythema/Ridging • 5-year survival rate ~34% Peau d’orange – 87% for other invasive breast cancers – Limited treatment options (typically estrogen receptornegative) and frequent resistance Adapted from: Woodward WA, et al., Radiation Oncology, 2009. Cancer Facts & Figures 2012, A.C. Society, Editor. 2012, American Cancer Society: Atlanta. Normal Resistance Response to to Therapy Primary Tumor Targeted Therapy (Lapatinib – inhibits EGFR) Growth Protein Signaling Stress Response Cancer Cancer Cell Cell Survival Death Aird KM, Ghanayem RB, et al. (2010) Mol Cancer Ther 9(5): 1432-1442. Hypothesis Primary Tumor • Patients continually exposed to ambient environmental chemicals • Can affect tumor signaling proteins • May lower treatment efficacy Environmental Agent Growth Protein Signaling Targeted Therapy (Lapatinib) Stress Response Cancer Cell Survival Demers A, Ayotte P, et al. 2000. Cancer Epidemiol Biomarkers Prev 9(2): 161-166. Duke, T. J., Jahed, N. C., et al. (2010) Oncology reports 24, 1277-1284. Gee, G. C., and Payne-Sturges, D. C. (2004) Environmental health perspectives 112, 1645-1653. ToxCast I Library (~300 compounds) High-Throughput MTT – Proliferation (~30 compounds) Automated liquid handling For assay assembly 340 nm light 520 nm light High-Content Imaging – Cell Behavior (~30 compounds) Energy transfer F Tb Protein Y Protein X Fluorescence-based Assay Design Functional Assays of Cancer Pathobiology (1 compound – BPA) Clonogenic/ Microcolony Growth AnchorageIndependent Growth Survival Signaling High-Throughput - 72h ToxCast I Library (~300 compounds) BPA Chlorothalonil % Proliferation 150 High-Throughput MTT – Proliferation (~30 compounds) 100 Cell Growth 50 0 2 3 4 log(Conc. [nM]) BPA Chlorothalonil 8 Nuclear Texture High-Content Imaging – Cell Behavior (~30 compounds) 6 4 2 Nuclear Count High-Content - 72h 150 100 BPA Chlorothalonil 50 0 0 2 3 4 log(Conc. [nM]) 2 3 4 log(Conc. [nM]) Cell Health Cell Growth Functional Assays of Cancer Pathobiology (1 compound – BPA) AnchorageIndependent Growth 150 * 100 100 nM Lapatinib 50 0 BPA (nM) 100 nM Lapatinib 1 - + 1 + 1 nM BPA + 100 nM Lapatinib Survival Signaling Colony Forming Efficiency (%) Colony Forming Efficiency (%) Clonogenic/ Microcolony Growth 150 2.5 μM Lapatinib 100 * 50 0 BPA (nM) 2.5 μM Lapatinib 500 - 500 - + + 500 nM BPA + 2.5 μM Lapatinib Functional Assays of Cancer Pathobiology (1 compound – BPA) AnchorageIndependent Growth Clonogenic/ Microcolony Growth 150 Quantification of AIG (%) * * 100 100 50 1 nM BPA 10 nM Staurosporine 0 Untreated Quantification of AIG (%) 150 Survival Signaling 50 0 BPA (nM) - Untreated BPA Lapatinib BPA + Lapatinib 1 100 nM Lapatinib Functional Assays of Cancer Pathobiology (1 compound – BPA) Clonogenic/ Microcolony Growth AnchorageIndependent Growth Survival Signaling BPA (30m) + - - + + + Lapatinib (2.5 μM) BPA (40 nM) pEGFR pEGFR 0.11 1.00 1.03 0.25 1.00 0.93 1.82 4.22 EGFR EGFR 1.28 1.00 0.86 1.20 1.00 1.05 0.77 0.70 GAPDH GAPDH Conclusions BPA identified from screen to increase inflammatory breast cancer (IBC) cell growth BPA inhibited anticancer efficacy of lapatinib, an FDA-approved therapy for IBC Environmental Biological Samples Samples – – Our Study (SUM149) • • BPA blocks main mechanism of lapatinib activity Occurs at doses found in healthy human blood samples and samples of patients receiving dialysis (medical tubing) Amniotic Fluid Normal Sera Saliva (post-dental sealing) Dialysis Patient Sera (Medical Tubing) Food containers Septage 1 nM 100 nM Hazardous Waste Landfill (median) 1 μM Clonogenic Growth (BPA+Lapatinib) 10 μM 100 μM High-content/MTT (BPA Alone) Anchorage-Independent Growth (BPA, BPA+Lapatinib) Signaling (BPA, BPA+Lapatinib) Kanno Y, Okada H et al. Therapeutic apheresis and dialysis. 2007;11(4):262-5. Vandenberg LN, Hauser R, et al. Reprod Toxicol. 2007;24(2):139-77. Acknowledgements • • Dr. Gayathri Devi Lab members • • • • • • Myron Evans Amy Stefanowicz John Davis Akshay Save Funding: – American Cancer Society – Viral Oncology Training Grant Collaborators: • H. Kim Lyerly, MD – Duke University • Kevin Williams, PhD – North Carolina Central University • Imran Shah, PhD – EPA, Durham, NC
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