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Moignard, V.* ; Macaulay, I.C.* ; Swiers, G.* ; Buettner, F. ; Schütte, J.* ; Calero-Nieto, F.J.* ; Kinston, S.* ; Joshi, A.* ; Hannah, R.* ; Theis, F.J. ; Jacobsen, S.E.* ; de Bruijn, M.F.* ; Göttgens, B.*

Characterization of transcriptional networks in blood stem and progenitor cells using high-throughput single-cell gene expression analysis.

Nat. Cell Biol. 15, 363-372 (2013)

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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.

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