title: Single-cell transcriptional analysis of normal, aberrant, and malignant hematopoiesis in zebrafish.
abstract: Hematopoiesis culminates in the production of functionally heterogeneous blood cell types. In zebrafish, the lack of cell surface antibodies has compelled researchers to use fluorescent transgenic reporter lines to label specific blood cell fractions. However, these approaches are limited by the availability of transgenic lines and fluorescent protein combinations that can be distinguished. Here, we have transcriptionally profiled single hematopoietic cells from zebrafish to define erythroid, myeloid, B, and T cell lineages. We also used our approach to identify hematopoietic stem and progenitor cells and a novel NK-lysin 4(+) cell type, representing a putative cytotoxic T/NK cell. Our platform also quantified hematopoietic defects in rag2(E450fs) mutant fish and showed that these fish have reduced T cells with a subsequent expansion of NK-lysin 4(+) cells and myeloid cells. These data suggest compensatory regulation of the innate immune system in rag2(E450fs) mutant zebrafish. Finally, analysis of Myc-induced T cell acute lymphoblastic leukemia showed that cells are arrested at the CD4(+)/CD8(+) cortical thymocyte stage and that a subset of leukemia cells inappropriately reexpress stem cell genes, including bmi1 and cmyb In total, our experiments provide new tools and biological insights into single-cell heterogeneity found in zebrafish blood and leukemia.
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129 dlangenau@mgh.harvard.edu femoore@mgh.harvard.edu.
authors.forename: Finola E
authors.initials: FE
authors.lastname: Moore
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: Elaine G
authors.initials: EG
authors.lastname: Garcia
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: Riadh
authors.initials: R
authors.lastname: Lobbardi
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114.
authors.forename: Esha
authors.initials: E
authors.lastname: Jain
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: Qin
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: John C
authors.initials: JC
authors.lastname: Moore
authors.affiliation: Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02115.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Cortes
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: Aleksey
authors.initials: A
authors.lastname: Molodtsov
authors.affiliation: Program in Molecular Medicine and Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA 01605.
authors.forename: Melissa
authors.initials: M
authors.lastname: Kasheta
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: Christina C
authors.initials: CC
authors.lastname: Luo
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: Amaris J
authors.initials: AJ
authors.lastname: Garcia
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129.
authors.forename: Ravi
authors.initials: R
authors.lastname: Mylvaganam
authors.affiliation: Department of Molecular Biomedical Sciences, North Carolina State University, Raleigh, NC 27607.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Yoder
authors.affiliation: Department of Pathology, University of Kentucky College of Medicine, Lexington, KY 40536 Department of Biochemistry, University of Kentucky College of Medicine, Lexington, KY 40536 Department of Molecular Biology, University of Kentucky College of Medicine, Lexington, KY 40536 Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY 40536.
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Blackburn
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114 Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114.
authors.forename: Ruslan I
authors.initials: RI
authors.lastname: Sadreyev
authors.affiliation: Program in Molecular Medicine and Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA 01605.
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Ceol
authors.affiliation: Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02115.
authors.forename: Trista E
authors.initials: TE
authors.lastname: North
authors.affiliation: Molecular Pathology, Massachusetts General Hospital, Charlestown, MA 02129 Cancer Center, Massachusetts General Hospital, Charlestown, MA 02129 Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114 Harvard Stem Cell Institute, Cambridge, MA 02139 dlangenau@mgh.harvard.edu femoore@mgh.harvard.edu.
authors.forename: David M
authors.initials: DM
authors.lastname: Langenau
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology; D006410:Hematopoiesis / Q000235:genetics / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000276:immunology* / Q000473:pathology; D007694:Killer Cells, Natural / Q000276:immunology / Q000473:pathology; D020125:Mutation, Missense; D014411:Neoplastic Stem Cells / Q000276:immunology* / Q000473:pathology; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology; D014158:Transcription, Genetic / Q000235:genetics / Q000276:immunology*; D015027:Zebrafish / Q000235:genetics / Q000276:immunology*; D029961:Zebrafish Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D004268:DNA-Binding Proteins; C583227:RAG2 protein, zebrafish; D029961:Zebrafish Proteins