title: A flexible reporter system for direct observation and isolation of cancer stem cells.
abstract: Many tumors are hierarchically organized with a minority cell population that has stem-like properties and enhanced ability to initiate tumorigenesis and drive therapeutic relapse. These cancer stem cells (CSCs) are typically identified by complex combinations of cell-surface markers that differ among tumor types. Here, we developed a flexible lentiviral-based reporter system that allows direct visualization of CSCs based on functional properties. The reporter responds to the core stem cell transcription factors OCT4 and SOX2, with further selectivity and kinetic resolution coming from use of a proteasome-targeting degron. Cancer cells marked by this reporter have the expected properties of self-renewal, generation of heterogeneous offspring, high tumor- and metastasis-initiating activity, and resistance to chemotherapeutics. With this approach, the spatial distribution of CSCs can be assessed in settings that retain microenvironmental and structural cues, and CSC plasticity and response to therapeutics can be monitored in real time.
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Binwu
authors.initials: B
authors.lastname: Tang
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Asaf
authors.initials: A
authors.lastname: Raviv
authors.affiliation: Protein Expression Laboratory, Advanced Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD 21701, USA.
authors.forename: Dominic
authors.initials: D
authors.lastname: Esposito
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Kathleen C
authors.initials: KC
authors.lastname: Flanders
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Daniel
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Bao Tram
authors.initials: BT
authors.lastname: Nghiem
authors.affiliation: Confocal Microscopy Core, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Garfield
authors.affiliation: Confocal Microscopy Core, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Langston
authors.initials: L
authors.lastname: Lim
authors.affiliation: Confocal Microscopy Core, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Poonam
authors.initials: P
authors.lastname: Mannan
authors.affiliation: Laboratory of Human Carcinogenesis, National Cancer Institute, Bethesda, MD 20892 USA.
authors.forename: Ana I
authors.initials: AI
authors.lastname: Robles
authors.affiliation: Department of Pathology, Suburban Hospital, Bethesda, MD 20814, USA.
authors.forename: William I
authors.initials: WI
authors.lastname: Smith
authors.affiliation: Program in Physical Biology, National Institute of Child Health and Human Development, Bethesda, MD 20892, USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: Zimmerberg
authors.affiliation: Program in Physical Biology, National Institute of Child Health and Human Development, Bethesda, MD 20892, USA.
authors.forename: Rea
authors.initials: R
authors.lastname: Ravin
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, Bethesda, MD 20892, USA. Electronic address: lw34g@nih.gov.
authors.forename: Lalage M
authors.initials: LM
authors.lastname: Wakefield
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D060049:Asymmetric Cell Division; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D058948:Cell Tracking; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D023061:Gene Order; D017930:Genes, Reporter; D005822:Genetic Vectors; D064593:Heterografts; D006801:Humans; D016130:Immunophenotyping; D051379:Mice; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D020218:Response Elements; D014157:Transcription Factors / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D014157:Transcription Factors