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Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development

ABSTRACT: We used single cell-RNA sequencing to investigate the expression and the heterogeneity of wild type and runx1-mutant cd41-GFPLow cd41GFP hematopoietic stem and progenitor cells at embryonic (2.5 days post fertilization, dpf) and larval stages (6, 10 16 dpf).

ORGANISM(S): Danio rerio

PROVIDER: GSE158099 | GEO | 2022/01/14

REPOSITORIES: GEO

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Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development

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Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development

Project description:Here we used RNA sequencing to characterize the transcriptional profile of the total RNA from c-Kit+ cells of Runx1 KO and control mice.

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Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development.

Project description:RUNX1 is essential for the generation of hematopoietic stem cells (HSCs). Runx1-null mouse embryos lack definitive hematopoiesis and die in mid-gestation. However, although zebrafish embryos with a runx1 W84X mutation have defects in early definitive hematopoiesis, some runx1W84X/W84X embryos can develop to fertile adults with blood cells of multilineages, raising the possibility that HSCs can emerge without RUNX1. Here, using 3 new zebrafish runx1-/- lines, we uncovered the compensatory mechanism for runx1-independent hematopoiesis. We show that, in the absence of a functional runx1, a cd41-green fluorescent protein (GFP)+ population of hematopoietic precursors still emerge from the hemogenic endothelium and can colonize the hematopoietic tissues of the mutant embryos. Single-cell RNA sequencing of the cd41-GFP+ cells identified a set of runx1-/--specific signature genes during hematopoiesis. Significantly, gata2b, which normally acts upstream of runx1 for the generation of HSCs, was increased in the cd41-GFP+ cells in runx1-/- embryos. Interestingly, genetic inactivation of both gata2b and its paralog gata2a did not affect hematopoiesis. However, knocking out runx1 and any 3 of the 4 alleles of gata2a and gata2b abolished definitive hematopoiesis. Gata2 expression was also upregulated in hematopoietic cells in Runx1-/- mice, suggesting the compensatory mechanism is conserved. Our findings indicate that RUNX1 and GATA2 serve redundant roles for HSC production, acting as each other's safeguard.

| S-EPMC9153008 | biostudies-literature

Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development [zebrafish bulk RNA-seq2]

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Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development

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Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development

Project description:This SuperSeries is composed of the SubSeries listed below.

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The transcriptional program controlled by Runx1 during early hematopoietic development (ChIP-seq data)

Project description:Transcription factors have long been recognised as powerful regulators of mammalian development, yet it is largely unknown how individual key regulators operate within wider regulatory networks. Here we have used a combination of global gene expression and chromatin-immunoprecipitation approaches across four ES-cell-derived populations of increasing haematopoietic potential to define the transcriptional programme controlled by Runx1, an essential regulator of blood cell specification. Integrated analysis of these complementary genome-wide datasets allowed us to construct a global regulatory network model, which suggested that core regulatory circuits are activated sequentially during blood specification, but will ultimately collaborate to control many haematopoietically expressed genes. Using the CD41/integrin alpha 2b gene as a model, cellular and in vivo studies showed that CD41 is controlled by both early and late circuits in fully specified blood cells, but initiation of CD41 expression critically depends on a later subcircuit driven by Runx1. Taken together, this study represents the first global analysis of the transcriptional programme controlled by any key haematopoietic regulator during the process of early blood cell specification. Moreover, the concept of interplay between sequentially deployed core regulatory circuits is likely to represent a design principle widely applicable to the transcriptional control of mammalian development. 4 samples: 3 samples of Runx1 transcription factor ChIP from the Runx1+/VE-cadherin+/CD41+ population, Runx1+/VE-cadherin+/CD41- population and Runx1+/VE-cadherin-/CD41+ population, and 1 Control (IgG).

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