Abstract

Breast Cancer Stem Cell Heterogeneity and Ultasensitive Mutation Detection using Sequencing

Anders Stahlberg, Principal Investigator, University Of Gothenburg

The identification of breast cancer cell subpopulations featuring truly malignant stem cell qualities is a challenge due to the complexity of the disease and lack of general markers. By combining extensive single-cell gene expression profiling with three functional strategies for cancer stem cell enrichment including anchorage-independent culture, hypoxia, and analyses of low-proliferative, label-retaining cells derived from mammospheres, we identified distinct stem cell clusters in breast cancer. Our findings allow us to understand the organization of breast cancers at the single-cell level, thereby permitting better identification and targeting of cancer stem cells. Detection of cell-free DNA in liquid biopsies offers great potential for use as a cancer biomarker. Here, we will also present an extremely simple and fast method for introduction of barcodes into DNA libraries, allowing ultrasensitive mutation detection. Our approach bridges the gap between the highly sensitive but specific capabilities of digital PCR, which only allows a limited number of variants to be analyzed, with the broad target capability of next-generation sequencing which traditionally lacks the sensitivity to detect rare variants.