Journal Article

GSK3β regulates epithelial-mesenchymal transition and cancer stem cell properties in triple-negative breast cancer

Den Hollander, Petra, Werden, Steven J., Chang, Jeffrey T., Pietila, Mika, Sarkar, Tapasree R., Mani, Sendurai A., Davies, Peter, Ramirez-Pena, Esmeralda, Miura, Noayuki, Zhao, Na, Soundararajan, Rama, Rosen, Jeffrey M., Vijay, Geraldine Vidhya, Shariati, Maryam, Gao, Ruli, Sobieski, Mary, Joseph, Robiya, Taube, Joseph H., Stephan, Clifford C., Sphyris, Nathalie, Toneff, Mike J.

2019

Overview

Background Triple-negative breast cancers (TNBCs), which lack receptors for estrogen, progesterone, and amplification of epidermal growth factor receptor 2, are highly aggressive. Consequently, patients diagnosed with TNBCs have reduced overall and disease-free survival rates compared to patients with other subtypes of breast cancer. TNBCs are characterized by the presence of cancer cells with mesenchymal properties, indicating that the epithelial to mesenchymal transition (EMT) plays a major role in the progression of this disease. The EMT program has also been implicated in chemoresistance, tumor recurrence, and induction of cancer stem cell (CSC) properties. Currently, there are no targeted therapies for TNBC, and hence, it is critical to identify the novel targets to treat TNBC. Methods A library of compounds was screened for their ability to inhibit EMT in cells with mesenchymal phenotype as assessed using the previously described Z-cad reporters. Of the several drugs tested, GSK3β inhibitors were identified as EMT inhibitors. The effects of GSK3β inhibitors on the properties of TNBC cells with a mesenchymal phenotype were assessed using qRT-PCR, flow cytometry, western blot, mammosphere, and migration and cell viability assays. Publicly available datasets also were analyzed to examine if the expression of GSK3β correlates with the overall survival of breast cancer patients. Results We identified a GSK3β inhibitor, BIO, in a drug screen as one of the most potent inhibitors of EMT. BIO and two other GSK3β inhibitors, TWS119 and LiCl, also decreased the expression of mesenchymal markers in several different cell lines with a mesenchymal phenotype. Further, inhibition of GSK3β reduced EMT-related migratory properties of cells with mesenchymal properties. To determine if GSK3β inhibitors target mesenchymal-like cells by affecting the CSC population, we employed mammosphere assays and profiled the stem cell-related cell surface marker CD44+/24− in cells after exposure to GSK3β inhibitors. We found that GSK3β inhibitors indeed decreased the CSC properties of cell types with mesenchymal properties. We treated cells with epithelial and mesenchymal properties with GSK3β inhibitors and found that GSK3β inhibitors selectively kill cells with mesenchymal attributes while sparing cells with epithelial properties. We analyzed patient data to identify genes predictive of poor clinical outcome that could serve as novel therapeutic targets for TNBC. The Wnt signaling pathway is critical to EMT, but among the various factors known to be involved in Wnt signaling, only the higher expression of GSK3β correlated with poorer overall patient survival. Conclusions Taken together, our data demonstrate that GSK3β is a potential target for TNBCs and suggest that GSK3β inhibitors could serve as selective inhibitors of EMT and CSC properties for the treatment of a subset of aggressive TNBC. GSK3β inhibitors should be tested for use in combination with standard-of-care drugs in preclinical TNBC models.

• Publisher: BioMed Central,Springer Nature B.V,BMC
• Subject: Binding sites / Biomedical and Life Sciences / Biomedicine / Bipolar disorder / Breast cancer / Cancer Research / Cancer stem cells (CSCs) / Cancer therapies / CD44 antigen / Cell adhesion & migration / Cell Line, Tumor / Cell migration / Cell Movement - drug effects / Cell surface / Cell viability / Chemoresistance / Data processing / Datasets as Topic / Disease / Drug development / Drug screening / Drug Screening Assays, Antitumor / Epidermal growth factor / Epithelial-mesenchymal transition (EMT) / Epithelial-Mesenchymal Transition - drug effects / Female / Flow cytometry / Gene expression / Genotype & phenotype / Glycogen Synthase Kinase 3 beta - antagonists & inhibitors / Glycogen Synthase Kinase 3 beta - metabolism / Glycogen synthase kinase β (GSK3β) / Humans / Inhibitory Concentration 50 / Kinases / Lithium chloride / Lithium Chloride - pharmacology / Lithium Chloride - therapeutic use / Medical prognosis / Mesenchyme / Metastasis / Neoplastic Stem Cells - drug effects / Neoplastic Stem Cells - pathology / Oncology / Patients / Phenotypes / Progesterone / Protein Kinase Inhibitors - pharmacology / Protein Kinase Inhibitors - therapeutic use / Pyrimidines - pharmacology / Pyrimidines - therapeutic use / Pyrroles - pharmacology / Pyrroles - therapeutic use / Research Article / Signal transduction / Stem cells / Surface markers / Surgical Oncology / Survival Analysis / Therapeutic applications / Triple Negative Breast Neoplasms - drug therapy / Triple Negative Breast Neoplasms - mortality / Triple Negative Breast Neoplasms - pathology / Triple-negative breast cancer (TNBC) / Tumors / Wnt protein / Wnt signaling / Wnt Signaling Pathway