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Activation of lineage competence in hemogenic endothelium precedes the formation of hematopoietic stem cell heterogeneity

ABSTRACT: Using a combination of single-cell multi-omics, lineage tracing and functional assays, we show that embryonic HSPCs are originated from heterogeneous hemogenic endothelial cells (HECs) during zebrafish embryogenesis.

ORGANISM(S): Danio rerio

PROVIDER: GSE186427 | GEO | 2023/03/24

REPOSITORIES: GEO

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Activation of lineage competence in hemogenic endothelium precedes the formation of hematopoietic stem cell heterogeneity [Zebrafish.scRNA-seq]

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Project description:CD49f has been used to define human hematopoietic stem cells (HSCs). Tracing the origin of CD49f+ HSCs shall enhance our understanding of the regulatory mechanisms and improve in vitro generation of HSCs. Here, we employed the hematopoietic differentiation system from human pluripotent stem cells and performed a computational trajectory analysis of the cells throughout this process based on integrated calculating the levels of 10 HSC markers. We observed two clades of cell populations in the trajectory tree, one of which represents the phenotypic CD49fHigh HSCs, and the other diverges only by low CD49f level. Live cell imaging revealed that CD49f expression is persistent during endothelial-to-hematopoietic transition, suggesting that the phenotypic CD49fHigh HSCs are pre-defined in hemogenic endothelial cells (HECs), while the concurrently emerged CD49fLow cells represent HSC-independent progenitors. Colony forming assays showed that the CD49fHigh cells have multilineage potential, whereas the CD49fLow cells lack lymphoid potential but show a strong erythroid preference. The aim of the experiment was to compare newly defined CD49fHigh and CD49fLow populations from hESCs-derived HSPCs and Hemogenic Endothelium.

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Project description:By tracing the VE-cadherin expression in the newborn bone marrow hematopoietic LSK (lineage minus/Sca-positive/Kit-positive) cells, we demonstrated that the late foetal/newborn BM hemogenic endothelial cells produce a small cohort of hematopoietic stem and progenitor cells (HSPCs) capable of circulating and colonizing the secondary haematopoietic organs. Phenotypic and functional analyses disclosed that BM endothelium-derived HSPCs are mainly Multipotent Progenitors (MPPs) and a few Hematopoietic Stem Cells. We used microarrays to detail the global programme of gene expression underlying the endothelial origin of LSK cells in the newborn bone marrow.

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Project description:Across species, hematopoietic stem and progenitor cells (HSPCs) arise during embryogenesis from a specialized arterial population, termed hemogenic endothelium. Here, we describe a mechanistic role for the epigenetic regulator, Enhancer of zeste homolog-1 (Ezh1) in vertebrate HSPC production via regulation of hemogenic commitment. Loss of ezh1 in zebrafish embryos favored acquisition of hemogenic (gata2b) and HSPC (runx1) fate at the expense of the arterial program (ephrinb2a, dll4). In contrast, ezh1 overexpression blocked hematopoietic progression via maintenance of arterial gene expression. The related polycomb group subunit, Ezh2, functioned in a non-redundant, sequential manner, whereby inhibition had no impact on arterial identity, but was capable of blocking ezh1-knockdown associated HSPC expansion. Single-cell RNA-seq across ezh1 genotypes revealed a dropout of ezh1+/- cells among arterial endothelium associated with positive regulation of gene transcription. Exploitation of Ezh1/2 modulation has potential functional relevance for improving in vitro HSPC differentiation from induced pluripotent stem cell sources.

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GATA2 is Dispensable for Generation of Hemogenic Endothelium But Required for Endothelial-to-Hematopoietic Transition

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