Therapeutic strategies that improve survival outcomes for advanced-stage breast cancers have proven a major clinical challenge. This is in part due to the lack of understanding of the underlying biology of disease progression. Cancer stem cells (CSCs) drive tumour initiation, metastatic spread and chemoresistance, hence eradicating these cells should lead to developing effective therapies. Triple-negative breast cancer (TNBC) is enriched in CSC subpopulations, as non-CSCs readily convert into CSCs through activation of non-genetic molecular mechanisms, which accounts for its aggressiveness. Through transcriptomic analysis of non-CSC and CSC cell subpopulations, we have defined the signalling network that governs the maintenance and formation of TNBC CSCs, and determined that the androgen receptor (AR) signalling is a major driver. In response to chemotherapy, AR activation switches cells into a cancer stem cell state, while AR antagonism suppress cell-state switching and cancer stem cell function. In vivo, the dual AR antagonist, seviteronel (VT-464/INO-464), enhances chemotherapy effectiveness, restricting tumour growth and metastasis development. Interestingly, we discovered that metastases and xenografted tumours with a CSC profile express high cytoplasmic AR levels. Analysis of 3 independent TNBC patient cohorts identified that cytoplasmic AR, which currently is not scored in the clinic, prognosticates poor survival in treatment-naïve patients and predicts lack of response to chemotherapy. In line with our findings, retrospective analysis of data from a Phase II clinical trial, showed that seviteronel treatment followed by chemotherapy improves survival for cytoplasmic-AR+ TNBC patients. Together, our work has shed light on the mechanisms that drive cancer stem cell biology, identifying a novel biomarker and therapeutic strategy with the potential to change clinical practice and improve TNBC patient’s outcomes. A clinical trial derived from this work has begun in Australia (NCT04947189).