Haematopoietic stem and progenitor cells from human pluripotent stem cells

Abstract

A variety of tissue lineages can be differentiated from pluripotent stem cells by mimicking embryonic development through stepwise exposure to morphogens, or by conversion of one differentiated cell type into another by enforced expression of master transcription factors. Here, to yield functional human haematopoietic stem cells, we perform morphogen-directed differentiation of human pluripotent stem cells into haemogenic endothelium followed by screening of 26 candidate haematopoietic stem-cell-specifying transcription factors for their capacity to promote multi-lineage haematopoietic engraftment in mouse hosts. We recover seven transcription factors (ERG, HOXA5, HOXA9, HOXA10, LCOR, RUNX1 and SPI1) that are sufficient to convert haemogenic endothelium into haematopoietic stem and progenitor cells that engraft myeloid, B and T cells in primary and secondary mouse recipients. Our combined approach of morphogen-driven differentiation and transcription-factor-mediated cell fate conversion produces haematopoietic stem and progenitor cells from pluripotent stem cells and holds promise for modelling haematopoietic disease in humanized mice and for therapeutic strategies in genetic blood disorders.

Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates

“…PU.1 cooperates with CEBPA and RUNX1 to command granulomonocytic lineage fates (20-27) -Pu.1-KO mice have reduced granulocytes and no monocytes (28,29); Cebpa-KO mice have no granulocytes (30); definitive hematopoiesis is abrogated in Runx1-KO mice (31); and ectopic expression of transcription factor ensembles containing PU.1 with CEBPA, or PU.1 with RUNX1, are sufficient to convert even fibroblasts into myeloid precursors/monocytes (29,(32)(33)(34)(35)(36). Functional compromise of this master circuit can hence be expected to impede myeloid differentiation, and accordingly, loss-of-function mutations/translocations of CEBPA and RUNX1 are highly recurrent in AML, a disease defined by myeloid differentiation arrest (37-39) (reviewed in ref.…”

Human haematopoietic stem cell development: from the embryo to the dish

“…Downregulation of HOXA5 and HOXA7 in human foetal liver HSCs supresses their capacity to self-renew and repopulate NSG recipients. Although, forced expression of HOXA5, HOXA7 and HOXA9 alone or in combination in HOXA-negative hPSC-derived progenitors was insufficient to convert the progenitors to HSCs (Dou et al, 2016), a recent study reported a greater degree of success by using a larger cohort of genes (Sugimura et al, 2017). Induced expression of seven transcription factors, notably including three HOXA genes, successfully converted hPSC-derived haematogenic endothelium into longterm repopulating HSCs (Sugimura et al, 2017).…”

“…Although, forced expression of HOXA5, HOXA7 and HOXA9 alone or in combination in HOXA-negative hPSC-derived progenitors was insufficient to convert the progenitors to HSCs (Dou et al, 2016), a recent study reported a greater degree of success by using a larger cohort of genes (Sugimura et al, 2017). Induced expression of seven transcription factors, notably including three HOXA genes, successfully converted hPSC-derived haematogenic endothelium into longterm repopulating HSCs (Sugimura et al, 2017). Finally, expression of four transcription factors (FOSB, GFI1, RUNX1 and SPI1) in human umbilical vein and dermal microvascular endothelial cells, or in adult mouse endothelial cells, were also sufficient to generate immunocompetent HSCs (Sandler et al, 2014;Lis et al, 2017).…”

“…Ultimately, the safest HSCs generated in the dish would likely be genetically unaltered cells, produced by exposure to external factors equivalent to the embryonic cues that drive HSC development. Here, the bottleneck seems to be identifying the combinations of soluble and cell-based factors and (Vereide et al, 2014); (2) (Kyba et al, 2002;Lu et al, 2016); (3) ; (4) (Elcheva et al, 2014); note that GATA2/ SCL combination induces erythro-megakaryocytic differentiation (not shown); (5) (Sandler et al, 2014); (6) (Riddell et al, 2014); (7) ; (8) (Batta et al, 2014); (9) (Cheng et al, 2016); (10) (Pulecio et al, 2014); (11) (Lis et al, 2017); (12) (Sugimura et al, 2017). CLPs, common lymphoid progenitors; CMPs, common myeloid progenitors; MPPs, multipotent progenitors.…”

The significance and application of vascular niche in the development and maintenance of hematopoietic stem cells

“…Neither method alone could induce HSCs. Therefore, I took an approach to combine these (Table 1) [12]. First, hemogenic endothelial cells were induced from human pluripotent stem cells by stepwise exposure to morphogens.…”