부산대학교 도서관

Keywords circulating tumor cells; circulating tumor cell clusters; single cell sequencing; bisulfite sequencing; RNA sequencing; stemness-associated transcription factors; proliferation-associated transcription factors; drug screen Highlights * Binding sites for OCT4, SOX2, NANOG, and SIN3A are hypomethylated in CTC clusters * CTC cluster hypomethylation profile correlates with a poor prognosis in breast cancer * Treatment with FDA-approved Na.sup.+/K.sup.+-ATPase inhibitors dissociates CTC clusters * Dissociation reverts the methylation profile of CTC clusters and suppresses metastasis Summary The ability of circulating tumor cells (CTCs) to form clusters has been linked to increased metastatic potential. Yet biological features and vulnerabilities of CTC clusters remain largely unknown. Here, we profile the DNA methylation landscape of single CTCs and CTC clusters from breast cancer patients and mouse models on a genome-wide scale. We find that binding sites for stemness- and proliferation-associated transcription factors are specifically hypomethylated in CTC clusters, including binding sites for OCT4, NANOG, SOX2, and SIN3A, paralleling embryonic stem cell biology. Among 2,486 FDA-approved compounds, we identify Na.sup.+/K.sup.+ ATPase inhibitors that enable the dissociation of CTC clusters into single cells, leading to DNA methylation remodeling at critical sites and metastasis suppression. Thus, our results link CTC clustering to specific changes in DNA methylation that promote stemness and metastasis and point to cluster-targeting compounds to suppress the spread of cancer.