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BMP signalling differentially regulates distinct haematopoietic stem cell types.

Crisan M, Kartalaei PS, Vink C, Yamada-Inagawa T, Bollerot K, van IJcken W, van der Linden R, de Sousa Lopes SM, Monteiro R, Mummery C, Dzierzak E - Nat Commun (2015)

Bottom Line: Adult haematopoiesis is the outcome of distinct haematopoietic stem cell (HSC) subtypes with self-renewable repopulating ability, but with different haematopoietic cell lineage outputs.Moreover, the two HSC types differ in intrinsic genetic programs, thus supporting a role for the BMP signalling axis in the regulation of HSC heterogeneity and lineage output.Our findings provide insight into the molecular control mechanisms that define HSC types and have important implications for reprogramming cells to HSC fate and treatments targeting distinct HSC types.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Cell Biology, Erasmus MC Stem Cell Institute, Erasmus Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands [2] University of Edinburgh, Centre for Inflammation Research, Queens Medical Research Institute, 47 Little France Crescent, Edinburgh EH16 4TJ, UK.

ABSTRACT
Adult haematopoiesis is the outcome of distinct haematopoietic stem cell (HSC) subtypes with self-renewable repopulating ability, but with different haematopoietic cell lineage outputs. The molecular basis for this heterogeneity is largely unknown. BMP signalling regulates HSCs as they are first generated in the aorta-gonad-mesonephros region, but at later developmental stages, its role in HSCs is controversial. Here we show that HSCs in murine fetal liver and the bone marrow are of two types that can be prospectively isolated--BMP activated and non-BMP activated. Clonal transplantation demonstrates that they have distinct haematopoietic lineage outputs. Moreover, the two HSC types differ in intrinsic genetic programs, thus supporting a role for the BMP signalling axis in the regulation of HSC heterogeneity and lineage output. Our findings provide insight into the molecular control mechanisms that define HSC types and have important implications for reprogramming cells to HSC fate and treatments targeting distinct HSC types.

No MeSH data available.


Aorta-gonad-mesonephros HSCs are BMP activated.(a) Scheme showing activation of the canonical BMP signalling pathway through phospo-Smad1/5 binding of the double BMP responsive element (BRE) and the resulting transcription of GFP. (b) Three-dimensional whole-mount image of an E10.5 immunostained BRE–GFP mouse embryo (37 somite pairs). Anti-CD31 (magenta), cKit (red) and GFP (green) antibody staining shows the predominantly ventral distribution of GFP+ cells in various cell types within the aorta and underlying mesenchymal cells. (c) High-magnification image of a transverse section of an E11 BRE–GFP dorsal aorta stained with anti-GFP-antibody and 4,6-diamidino-2-phenylindole (DAPI). Intra-aortic hematopoietic clusters contain GFP+ and GFP− cells. Some endothelial cells are GFP+. (d) Representative FACS plot with side scatter on the y axis showing percentage of E11 AGM GFP+ cells (see Supplementary Fig. 1A for control). (e) Percentage donor cell chimerism in the peripheral blood of adult irradiated transplant recipients at 4 months after injection of E11 AGM GFP+ and GFP− cells (2–4.5 embryo equivalents (ee) of AGM cells injected per mouse; n=3 independent transplantation experiments). *P=0.05 by z-Test for proportions.
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f1: Aorta-gonad-mesonephros HSCs are BMP activated.(a) Scheme showing activation of the canonical BMP signalling pathway through phospo-Smad1/5 binding of the double BMP responsive element (BRE) and the resulting transcription of GFP. (b) Three-dimensional whole-mount image of an E10.5 immunostained BRE–GFP mouse embryo (37 somite pairs). Anti-CD31 (magenta), cKit (red) and GFP (green) antibody staining shows the predominantly ventral distribution of GFP+ cells in various cell types within the aorta and underlying mesenchymal cells. (c) High-magnification image of a transverse section of an E11 BRE–GFP dorsal aorta stained with anti-GFP-antibody and 4,6-diamidino-2-phenylindole (DAPI). Intra-aortic hematopoietic clusters contain GFP+ and GFP− cells. Some endothelial cells are GFP+. (d) Representative FACS plot with side scatter on the y axis showing percentage of E11 AGM GFP+ cells (see Supplementary Fig. 1A for control). (e) Percentage donor cell chimerism in the peripheral blood of adult irradiated transplant recipients at 4 months after injection of E11 AGM GFP+ and GFP− cells (2–4.5 embryo equivalents (ee) of AGM cells injected per mouse; n=3 independent transplantation experiments). *P=0.05 by z-Test for proportions.

Mentions: The localized production of BMP4 in the AGM101112 and BMPR2 expression by enriched AGM haematopoietic progenitors and stem cells (HPSCs)10 suggests that BMP may act directly on HSCs. We examined the BMP activation status of HSCs during ontogeny in BRE–GFP transgenic mice. GFP is expressed in BRE–GFP mice when BMP and the BMP receptor signal through Smad1/5 to activate transcription from the BRE sequence (Fig. 1a). GFP expression denotes BMP activation at the moment of cell observation/isolation and does not represent previous BMP activation history. Importantly, all BRE–GFP-expressing cells also express pSmad1/5/8 (ref. 13). Three-dimensional imaging of whole-mount immunostained transgenic E10.5 embryos shows that BMP-activated (GFP+) cells are predominantly distributed in the ventral aspect of the aorta (lumenal hematopoietic, endothelial and underlying mesenchymal cells; Fig. 1b). HPSCs are known to reside in the haematopoietic clusters closely associated with the aorta141516. Some cluster cells are GFP+ (Fig. 1c), suggesting heterogeneity in BMP activation within the HPSC compartment at the time of HSC generation. To test the BMP activation status of HSCs, E11 AGM GFP+ and GFP− cells were sorted (6.2±3.1%(mean±s.d.) of E11 AGM cells are GFP+; Fig. 1d, Supplementary Fig. 1A) and transplanted into adult irradiated recipient mice. All AGM HSCs were found in the BMP-activated fraction (Fig. 1e): 3 out of 7 recipients receiving GFP+ AGM cells were long term, high level repopulated (>10% chimerism) by donor cells, whereas no recipients (0 out of 7) receiving GFP− AGM cells were donor engrafted (P=0.05), even when high embryo equivalents of cells were injected. These data show that BMP signalling is activated in HSCs when they are first detected in the AGM.


BMP signalling differentially regulates distinct haematopoietic stem cell types.

Crisan M, Kartalaei PS, Vink C, Yamada-Inagawa T, Bollerot K, van IJcken W, van der Linden R, de Sousa Lopes SM, Monteiro R, Mummery C, Dzierzak E - Nat Commun (2015)

Aorta-gonad-mesonephros HSCs are BMP activated.(a) Scheme showing activation of the canonical BMP signalling pathway through phospo-Smad1/5 binding of the double BMP responsive element (BRE) and the resulting transcription of GFP. (b) Three-dimensional whole-mount image of an E10.5 immunostained BRE–GFP mouse embryo (37 somite pairs). Anti-CD31 (magenta), cKit (red) and GFP (green) antibody staining shows the predominantly ventral distribution of GFP+ cells in various cell types within the aorta and underlying mesenchymal cells. (c) High-magnification image of a transverse section of an E11 BRE–GFP dorsal aorta stained with anti-GFP-antibody and 4,6-diamidino-2-phenylindole (DAPI). Intra-aortic hematopoietic clusters contain GFP+ and GFP− cells. Some endothelial cells are GFP+. (d) Representative FACS plot with side scatter on the y axis showing percentage of E11 AGM GFP+ cells (see Supplementary Fig. 1A for control). (e) Percentage donor cell chimerism in the peripheral blood of adult irradiated transplant recipients at 4 months after injection of E11 AGM GFP+ and GFP− cells (2–4.5 embryo equivalents (ee) of AGM cells injected per mouse; n=3 independent transplantation experiments). *P=0.05 by z-Test for proportions.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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f1: Aorta-gonad-mesonephros HSCs are BMP activated.(a) Scheme showing activation of the canonical BMP signalling pathway through phospo-Smad1/5 binding of the double BMP responsive element (BRE) and the resulting transcription of GFP. (b) Three-dimensional whole-mount image of an E10.5 immunostained BRE–GFP mouse embryo (37 somite pairs). Anti-CD31 (magenta), cKit (red) and GFP (green) antibody staining shows the predominantly ventral distribution of GFP+ cells in various cell types within the aorta and underlying mesenchymal cells. (c) High-magnification image of a transverse section of an E11 BRE–GFP dorsal aorta stained with anti-GFP-antibody and 4,6-diamidino-2-phenylindole (DAPI). Intra-aortic hematopoietic clusters contain GFP+ and GFP− cells. Some endothelial cells are GFP+. (d) Representative FACS plot with side scatter on the y axis showing percentage of E11 AGM GFP+ cells (see Supplementary Fig. 1A for control). (e) Percentage donor cell chimerism in the peripheral blood of adult irradiated transplant recipients at 4 months after injection of E11 AGM GFP+ and GFP− cells (2–4.5 embryo equivalents (ee) of AGM cells injected per mouse; n=3 independent transplantation experiments). *P=0.05 by z-Test for proportions.
Mentions: The localized production of BMP4 in the AGM101112 and BMPR2 expression by enriched AGM haematopoietic progenitors and stem cells (HPSCs)10 suggests that BMP may act directly on HSCs. We examined the BMP activation status of HSCs during ontogeny in BRE–GFP transgenic mice. GFP is expressed in BRE–GFP mice when BMP and the BMP receptor signal through Smad1/5 to activate transcription from the BRE sequence (Fig. 1a). GFP expression denotes BMP activation at the moment of cell observation/isolation and does not represent previous BMP activation history. Importantly, all BRE–GFP-expressing cells also express pSmad1/5/8 (ref. 13). Three-dimensional imaging of whole-mount immunostained transgenic E10.5 embryos shows that BMP-activated (GFP+) cells are predominantly distributed in the ventral aspect of the aorta (lumenal hematopoietic, endothelial and underlying mesenchymal cells; Fig. 1b). HPSCs are known to reside in the haematopoietic clusters closely associated with the aorta141516. Some cluster cells are GFP+ (Fig. 1c), suggesting heterogeneity in BMP activation within the HPSC compartment at the time of HSC generation. To test the BMP activation status of HSCs, E11 AGM GFP+ and GFP− cells were sorted (6.2±3.1%(mean±s.d.) of E11 AGM cells are GFP+; Fig. 1d, Supplementary Fig. 1A) and transplanted into adult irradiated recipient mice. All AGM HSCs were found in the BMP-activated fraction (Fig. 1e): 3 out of 7 recipients receiving GFP+ AGM cells were long term, high level repopulated (>10% chimerism) by donor cells, whereas no recipients (0 out of 7) receiving GFP− AGM cells were donor engrafted (P=0.05), even when high embryo equivalents of cells were injected. These data show that BMP signalling is activated in HSCs when they are first detected in the AGM.

Bottom Line: Adult haematopoiesis is the outcome of distinct haematopoietic stem cell (HSC) subtypes with self-renewable repopulating ability, but with different haematopoietic cell lineage outputs.Moreover, the two HSC types differ in intrinsic genetic programs, thus supporting a role for the BMP signalling axis in the regulation of HSC heterogeneity and lineage output.Our findings provide insight into the molecular control mechanisms that define HSC types and have important implications for reprogramming cells to HSC fate and treatments targeting distinct HSC types.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Cell Biology, Erasmus MC Stem Cell Institute, Erasmus Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands [2] University of Edinburgh, Centre for Inflammation Research, Queens Medical Research Institute, 47 Little France Crescent, Edinburgh EH16 4TJ, UK.

ABSTRACT
Adult haematopoiesis is the outcome of distinct haematopoietic stem cell (HSC) subtypes with self-renewable repopulating ability, but with different haematopoietic cell lineage outputs. The molecular basis for this heterogeneity is largely unknown. BMP signalling regulates HSCs as they are first generated in the aorta-gonad-mesonephros region, but at later developmental stages, its role in HSCs is controversial. Here we show that HSCs in murine fetal liver and the bone marrow are of two types that can be prospectively isolated--BMP activated and non-BMP activated. Clonal transplantation demonstrates that they have distinct haematopoietic lineage outputs. Moreover, the two HSC types differ in intrinsic genetic programs, thus supporting a role for the BMP signalling axis in the regulation of HSC heterogeneity and lineage output. Our findings provide insight into the molecular control mechanisms that define HSC types and have important implications for reprogramming cells to HSC fate and treatments targeting distinct HSC types.

No MeSH data available.