title: Characterization of transcriptional networks in blood stem and progenitor cells using high-throughput single-cell gene expression analysis.
abstract: Cellular decision-making is mediated by a complex interplay of external stimuli with the intracellular environment, in particular transcription factor regulatory networks. Here we have determined the expression of a network of 18 key haematopoietic transcription factors in 597 single primary blood stem and progenitor cells isolated from mouse bone marrow. We demonstrate that different stem/progenitor populations are characterized by distinctive transcription factor expression states, and through comprehensive bioinformatic analysis reveal positively and negatively correlated transcription factor pairings, including previously unrecognized relationships between Gata2, Gfi1 and Gfi1b. Validation using transcriptional and transgenic assays confirmed direct regulatory interactions consistent with a regulatory triad in immature blood stem cells, where Gata2 may function to modulate cross-inhibition between Gfi1 and Gfi1b. Single-cell expression profiling therefore identifies network states and allows reconstruction of network hierarchies involved in controlling stem cell fate choices, and provides a blueprint for studying both normal development and human disease.
authors.affiliation: Department of Haematology, Wellcome Trust and MRC Cambridge Stem Cell Institute and Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.
authors.forename: Victoria
authors.initials: V
authors.lastname: Moignard
authors.forename: Iain C
authors.initials: IC
authors.lastname: Macaulay
authors.forename: Gemma
authors.initials: G
authors.lastname: Swiers
authors.forename: Florian
authors.initials: F
authors.lastname: Buettner
authors.forename: Judith
authors.initials: J
authors.lastname: Schütte
authors.forename: Fernando J
authors.initials: FJ
authors.lastname: Calero-Nieto
authors.forename: Sarah
authors.initials: S
authors.lastname: Kinston
authors.forename: Anagha
authors.initials: A
authors.lastname: Joshi
authors.forename: Rebecca
authors.initials: R
authors.lastname: Hannah
authors.forename: Fabian J
authors.initials: FJ
authors.lastname: Theis
authors.forename: Sten Eirik
authors.initials: SE
authors.lastname: Jacobsen
authors.forename: Marella F
authors.initials: MF
authors.lastname: de Bruijn
authors.forename: Berthold
authors.initials: B
authors.lastname: Göttgens
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics*; D001853:Bone Marrow / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059010:Single-Cell Analysis
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger; D008156:Luciferases