Download Haematopoietic stem cells, niches and differentiation

Haematopoietic stem cells, niches
and differentiation pathways
Thomas Graf and Andreas Trumpp
Haematopoietic stem cells (HSCs) continuously replenish blood cells that are lost by
attrition or tissue damage. They are capable of self-renewal and are currently the only
adult stem-cell type routinely used in clinical settings to replace lost cells. HSCs are mostly
quiescent but can be mobilized from their niche to proliferate and differentiate into
lineages of the innate and adaptive immune system, as well as into red blood cells and
platelets. Cell-fate decisions are initiated and maintained by specific combinations of
transcription factors, the activity of which is orchestrated by extrinsic and intrinsic signals.
The study of changes in regulatory networks during haematopoietic differentiation has
long served as a paradigm for basic processes of cell-fate specification and its aberrations,
such as those that occur in leukaemia. The easy accessibility and transplantability of normal
and leukaemic haematopoietic cells has led to the discovery of cytokines, oncogenes and
cancer stem cells and to some of the most celebrated successes of targeted drug design.
Guide through the poster
Placenta
HSC niches
Yolk sac
Bone marrow
Sinusoid
Mesenchymal HSC precursors
HSCs are transiently present
in the placenta (E11–E15)
Bone marrow
Thymic rudiment
Mouse embryo
Endothelial cell
CXCL12-expressing
reticular cell
From E10 onwards, AGM-derived
HSCs colonize the fetal liver,
where they expand and mature.
Approximately at birth, HSCs start
to migrate to the bone marrow,
which becomes the main site of
haematopoiesis in the adult.
HSCs are generated in the
AGM between E8.5 and E.11.5
TPO
GATA1
FOG1
ETS1
Vascular niche
Eosinophil
Erythrocyte
Megakaryocyte
EPO
GATA1
FOG1
EKLF
IL-5
GATA2
C/EBPα
Fetal liver
F-HSC
ST-HSC
LT-HSC
IL-3
MPP
ECM
MBT
ID2
SPIB
IL-7
SCF
FLT3L
Low O2
MT
NK
Neutrophil
CXCL12expressing
reticular cell
Macrophage Megakaryocyte
T-cell development
in the thymus
Sonic
hedgehog
Phenotypes of HSCs
Mouse: Lin–Sca1+Kit+CD34–CD48–CD150+
Human: LIN–CD34+CD38–THY1lowKIT+
Patched
Notchligand
DLL4producing
cortical
epithelial
cell
Effect on self-renewal capacity
+ (Promote HSC maintenance and/or quiescence): BMI1, FOXO, GFI1, STAT5, ATM, p57, HOXB4
– (Promote HSC proliferation and/or differentiation): cMYC, cMYB, JUNB, β-catenin, p18, p27
Adhesion
Self-renewal,
Survival
Dormancy,
adhesion
Self-renewal
Lodging
CXCL12
Migration
RAC
KIT
CD44
IgM+
NK cell
Immature
B cell (T1)
Fetal precursor
IL-7
RAG
SOX13
B-1a/b cell
(In peritoneum
as well as in spleen)
γδ T cell
DN3
TCRβ GATA3
Notch1 RAG
IgMhi
IgDlow
Antigen
stimulation,
helper T-cells
OPN
BMP
ECM
Migration
TCF
E2A
?
Calcium
receptor
β-catenin
Hyaluronic
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Memory T cell
Effector T cell
Abbreviations
AGM, aorta–gonad–mesonephros; ANG1, angiopoietin 1;
APRIL, a proliferation-inducing ligand; ATM, ataxia-telangiectasia
mutated; BAFF, B-cell-activating factor; BCL, B-cell lymphoma;
BCR, B-cell receptor; BLIMP1, B-lymphocyte-induced
maturation protein 1; BMP, bone morphogenetic protein;
BMPR1A, bone morphogenetic protein receptor, type 1 A;
C/EBPα, CCAAT/enhancer-binding protein α; CLP, common
lymphoid progenitor; CMP, common myeloid progenitor;
CXCL12, CXC-chemokine ligand 12; CXCR4, CXC-chemokine
receptor 4; DC, dendritic cell; DLL4, delta-like ligand 4;
DN, double negative; DP, double positive; E2A, transcription
factor E2A; EBF, early B-cell factor; ECM, extracellular matrix;
EGR1, early-growth response; EKLF, erythroid Kruppel-like
factor; EPO, erythropoietin; ETP, early T-cell-lineage progenitor;
TH1 cell
IL-6
BLIMP1
STAT3
NF-κB
IL-6
TGFβ
RORγt
Plasma
cell
IL-4
GATA3
STAT6
PTHR
www.abcam.com/stemcells
BAFF
Strong BCR
signal
?
?
ANG1
BMPR1A
IgMlow
IgD+
Mature
B cell (T2)
Notch2
BAFF
Weak BCR
signal
VLA4
Cadherins
CXCL12
CD44
Notch
Cadherins
VLA5
TIE2
Self-renewal
Non-canonical
WNT signalling
?
CXCR4
www.abcam.com/stemcells
Ikaros
ETS1
PU.1
DN4
MPL
Osteoblast
IL-15
IL-2
ID2
TCRβ
GATA3
RAG
Survival
MCL1
Self-renewal
Dormancy
Immature B cell
DN2
Smoothened
TPO
Pre-B cell
CXCL12-expressing
reticular cell
IL-7
TCF
FLT3L PU.1
E2A
GATA3
Nucleus
Dormant HSC
Pro-B cell
IgM
BCR
RAG
B-cell development
in the spleen
Notch1
Ikaros
ETP/
DN1
B cell
CXCL12expressing
reticular cell
Pre/proB cell
preBCR
RAG
TSP
Osteoblast
Bone
Macrophage/
neutrophil
DC
CXCL12
PAX5
NK cell
Osteocyte
T cell
DC
FLT3L E2A
CXCL12 EBF
Differentiation,
migration and
expansion
Dormant
LT-HSC
MB
GM-CSF
IL-4
DC
CLP
LMPP
MPP
Monocyte/
macrophage
ID2
SPIB
Fibroblast
Endosteal niche
M-CSF PU.1hi
EGR1/2 IRF8
MAFB C/EBPα
GMP
Mast cell
Dividing HSC
Neutrophil
PU.1hi
C/EBPα
CMP
FLT3L
SCF
PU.1
Homeostasis (rare)
Injury (frequent)
Erythrocyte
IL-3
GATA2
C/EBPα
MEP
Activated
LT-HSC
G-CSF
C/EBPα
GFI1
Basophil
PAX5
BCL6
IgG+
Memory B cell
TH2 cell
TH17 cell
ETS1, v-ets erythroblastosis virus E26 oncogene homologue 1;
F-HSC, fetal HSC; FLT3L, FMS-related-tyrosine-kinase-3 ligand;
FOG1, friend of GATA 1; FOX, forkhead box; GATA1, GATAbinding protein 1; G-CSF, granulocyte colony-stimulating factor;
GFI1, growth-factor independent 1; GM-CSF, granulocyte/
macrophage CSF; GMP, granulocyte/macrophage progenitor;
HOXB4, homeobox B4; HSC, haematopoietic stem cell; ICAM1,
intercellular adhesion molecule 1; ID2, inhibitor of DNA
binding 2; IFNγ, interferon-γ; IL, interleukin; IRF8, interferonregulatory factor 8; LIN, lineage; LMPP, lymphoid primed MPP;
LRP5, low-density-lipoprotein-receptor-related protein 5;
LT-HSC, long-term repopulating HSC; MB, myeloid/B-cell
precursor; MBT, myeloid, T- and B-cell precursor; MCL1,
myeloid-cell leukaemia sequence 1; M-CSF, macrophage
colony-stimulating factor; MEP, megakaryocyte/erythroid
progenitor; MPL, myeloproliferative leukaemia virus oncogene;
MPP, multipotent progenitor; MT, myeloid/T-cell precursor;
NF-κB, nuclear factor-κB; NK, natural killer; OPN, osteopontin;
PAX5, paired box gene/protein 5; PTH, parathyroid hormone;
PTHR, parathyroid hormone receptor; PU.1, transcription factor
PU.1; RAG, recombination-activating gene 1; RORγt, retinoicacid-receptor-related orphan receptor-γt; RUNX, runt-related
transcription factor; SCA1, stem-cell antigen 1; SCF, stemcell factor; SOX13, SRY box 13; SPIB, Spi-B transcription
factor; STAT, signal transducer and activator of transcription;
ST-HSC, short-term repopulating HSC; T1, transitional B cell 1;
TCF, T-cell factor; TCR, T-cell receptor; TGFβ, transforming
growth factor-β; TH, T helper; ThPOK, T helper-inducing
POZ/Kruppel factor; TIE2, tyrosine kinase receptor 2;
TPO, thrombopoietin; TReg, T regulatory; TSP, thymus-seeding
progenitor; VCAM1; vascular cell-adhesion molecule 1;
VLA, very late antigen.
The upper left region of the poster shows a mouse
embryo with its haematopoietic anlagen. Although the
extent of yolk-sac contribution to the formation of adult
HSCs is controversial, most HSCs are probably formed in
the aorta–gonad–mesonephros (AGM) region. From here,
stem cells seed the fetal liver, where they expand and
differentiate into myeloid and lymphoid lineages.
Subsequently, HSCs exit the liver and populate
specialized bone-marrow microenvironments (niches) in
which they gradually become quiescent. As shown in the
lower left panel, dormant HSCs are mostly located close
to the lining of the trabecular bone, the endosteal niche.
This niche contains osteoblasts and CXC-chemokine
ligand 12 (CXCL12)-expressing reticular (CAR) cells.
Several cell adhesion molecules, together with shortrange signalling molecules, control HSC lodging and
maintenance (lower left panel). In response to specific
signals, it is thought that HSCs are recruited to vascular
niches — that are comprised of CAR cells and
fenestrated sinusoidal endothelial cells — where they
begin to divide, differentiate and enter the circulation.
Bone-marrow-derived thymus-seeding progenitors
(TSPs) seed the thymus and become committed to the
T-cell lineage (lower right panel). In the thymus, the cells
travel from the cortex through the subcapsular zone to
the medulla, encountering different epithelial niches
that guide them through several developmental stages.
Finally, CD4+ and CD8+ T cells enter the circulation and
differentiate into mature effector and helper T cells in
the spleen and lymph nodes. B-lineage cells become
committed in the bone marrow and, after immature
B cells enter the circulation and home to the spleen,
they mature into antibody-secreting memory B cells and
plasma cells (lower right panel).
The differentiation pathways are shown beginning
with HSCs and ending with mature cells. Dotted lines
represent transitions in which the origin of the more
mature cell is not clear. Arrows indicate points of
migration of circulating cells (with the exception of the
HSC synapse panel). It is likely that the early stages of
HSC development represent a continuum. There are still
uncertainties about several parts of the pathways. The
poster shows multipotent progenitors (MPPs) branching
into megakaryocyte/erythroid progenitors (MEPs) and
lymphoid-primed multipotent progenitors (LMPPs).
LMPPs either become common lymphoid progenitors
(CLPs) or common myeloid progenitors (CMPs) and
granulocyte/macrophage progenitors (GMPs). However,
GMPs can also generate eosinophils, basophils and mast
cells and it is possible that more than one pathway to
granulocytes and macrophages exists. Finally, there are
controversies about the developmental origin of the
different types of dendritic cells and B cells. The poster
also shows lineage-instructive signals such as chemokines,
cytokines and transcription factors at the points where
they are required. The definition of a lineage-instructive
factor is complex and is ideally based on gene ablation,
cell-type-specific gene inactivation and gain-of-function
experiments. Since this applies only to a fraction of the
factors shown here, the information provided is
somewhat speculative. Also, only transcription factors
that act dominantly during lineage commitment are
listed, but not those that need to be silenced.
Thomas Graf is an ICREA investigator at the Centre for Genomic Regulation,
Dr Aiguader 88, E08003 Barcelona, Spain. e-mail: [email protected]
Andreas Trumpp is at the Swiss Institute for Experimental Cancer Research (ISREC)
and the Ecole Polytechnique Fédérale de Lausanne (EPFL), Chemin des Boveresses
155, 1066 Epalinges, Lausanne, Switzerland. e-mail: [email protected]
Supplementary information (glossary terms and background reading) is associated
with the online version of this poster. www.nature.com/nri/posters/hsc
Edited by Elaine Bell; copy edited by Marta Tufet; designed by Neil Smith and
Simon Fenwick. © 2007 Nature Publishing Group.
Glossary
Haematopoietic stem cells
(HSCs). Cells that are capable of reconstituting all lineages of the
haematopoietic system after transplantation into lethally irradiated
mice. Long-term repopulating HSCs (LT-HSCs) do this life-long and
after secondary transplantations into irradiated hosts, whereas
short-term repopulating HSCs (ST-HSCs) only show multilineage
repopulation for a few weeks. LT- and ST-HSCs can be identified
and isolated by flow cytometry on the basis of their expression of
specific cell-surface antigens.
Lineage commitment
Also known as cell-fate determination; a process whereby HSCs
become specialized while losing their self-renewal capacity.
Progenitor cells
Also known as precursor cells; are intermediates between HSCs and
differentiated cells with restricted differentiation potential, high
proliferative capacity, but little or no self-renewal capacity. They
are probably equivalent to the transit amplifying cells of other adult
stem-cell systems.
Myeloid cells
A term variously used to define non-lymphoid cells or cells of the
monocyte–macrophage compartment (known as myelomonocytic
cells).
Granulocytes
A group of cells including neutrophils, eosinophils, and basophils,
but the term is often used for neutrophils only.
Bone-marrow and/or HSC transplantation
A procedure involving injection of HSCs or bone marrow into
irradiated hosts to determine the cell’s biological potential. In the
clinic it is a life-saving procedure after chemotherapy or radiation
therapy that destroys the haematopoietic system. Bone-marrow
transplantation typically requires immunosuppression to prevent
graft rejection due to tissue mismatch.
Cord-blood stem cells
These are fetal HSCs present in the umbilical cord of newborns.
They are used clinically as an alternative to bone-marrow-derived
HSCs. If stored frozen and thawed years later, they can be useful for
regenerating the haematopoietic system of the donor, obviating the
danger of tissue rejection of mismatched transplants.
Dormancy and mobilization
Although HSCs self-renew, they are mostly quiescent, dividing
approximately once a month. Proliferation can be activated in
response to injury or injection of cytokines, such as granulocyte
colony-stimulating factor (G-CSF). G-CSF also induces the exit
of HSCs from their niches and mobilization of the cells into the
circulation, a procedure that is used clinically to obtain HSCs from a
patient’s blood.
Cytokines and chemokines
Secreted proteins that stimulate cell growth, survival and
differentiation.
Stem-cell niches
Specialized microenvironments that control stem-cell dormancy
and the balance between stem-cell self-renewal and differentiation.
Colony-forming assay
If seeded in semi-solid medium, blood-cell precursors form colonies
in the presence of appropriate cytokines. This assay is widely
used to define a cell’s differentiation potential and to identify
biologically active molecules.
Plasticity
The capacity of defined cell types to acquire new identities. The
extent of plasticity of HSCs and haematopoietic progenitors under
physiological conditions is controversial and probably quite limited.
However, re-specification of cell fate can be induced by enforced
transcription factor expression, cell fusion or nuclear transfer.
Cancer and/or leukaemia stem cells
These are self-renewing cells capable of generating leukaemia
after transplantation. They can give rise to more differentiated
cells comprising the bulk of the leukaemia. Cancer stem cells,
first identified in acute myeloid leukaemia, are critical targets for
intervention.
Self-renewal
The ability of cells to repeatedly generate at least one identical
daughter cell.
Lineage priming
The promiscuous expression of myeloid–erythroid lineage
associated markers in HSCs.
Background reading
Wilson, A. & Trumpp, A. Bone-marrow haematopoietic-stem-cell
niches. Nature Rev. Immunol. 6, 93–106 (2006).
Takahama Y. Journey through the thymus: stromal guides for T-cell
development and selection. Nature Rev. Immunol. 6, 127–135 (2006).
Rothenberg E.V. Cell lineage regulators in B and T cell development.
Nature Immunol. 8, 441–444 (2007).
Cumano, A. & Godin, I. Ontogeny of the hematopoietic system.
Annu Rev. Immunol. 25, 745–785 (2007).
Laiosa C.V., Stadtfeld, M. & Graf, T. Determinants of lymphoidmyeloid lineage diversification. Annu. Rev. Immunol. 24, 705–738
(2006).
Nagasawa T. Microenvironmental niches in the bone marrow
required for B-cell development. Nature Rev. Immunol. 6, 107–116
(2006).