Haemopoiesis

CHAPTER 1
Haemopoiesis

• blood cell formation

• formation of red cells (erythropoiesis), granulocytes and monocytes
(myelopoiesis) and platelets (thrombopoiesis) are also discussed.
Site of haemopoiesis
• infancy
• all the bone marrow is haemopoietic
• childhood
• there is progressive fatty replacement of
marrow throughout the long bones so
that in
• adult life
• Haemopoietic marrow is confined to the
central skeleton and proximal ends of
the femurs and humeri (Table 1.1).
 • Even in these haemopoietic areas, approximately 50% of the marrow consists
of fat
• The remaining fatty marrow is capable of reversion
• to haemopoiesis
• In many diseases there is also expansion of haemopoiesis down the
long bones.
• The liver and spleen can resume their fetal haemopoietic role
(‘extramedullary haemopoiesis’).
Haemopoietic stem and
progenitor cells
• Haemopoiesis starts
with a pluripotential
stem cell that can by
asymmetric cell division
self‐renew but also give
rise to the separate cell
lineages.
Cell differentiation occurs
from the stem cell via
committed haemopoietic
progenitors which are
restricted in their
developmental potential
 Bone marrow cells are increasingly differentiated and lose the
The stem cell has the capability for self‐renewal capacity for self‐renewal as they mature.

(b) A single stem cell gives rise, after multiple cell divisions
(shown by vertical lines), to >106 mature cells
• With aging, the number of stem cells falls
• Stem cells also accumulate genetic mutations with age, an average of
8 at age 60
 Bone marrow stroma
• Forms a suitable
environment for stem
cell survival, renewal
and formation of
differentiated
progenitor cells
• Composed of stromal
cells and microvascular
network
 Bone marrow stroma
• Stromal cells: connective tissue cells of any organ
• Adipocytes, fibroblasts, osteoblasts, endothelial cells, macrophages
 Bone marrow stroma
• Stromal cells:
• These secrete extracellular molecules such as collagen,
glycoproteins (fibronectin and thrombospondin);
glycoproteins (hyaluronic acid and chrondroitin) to form the
extracellular matrix
• Secrete growth factors needed for stem cell renewal
• Major source of growth factors except for erythropoietin
(kidney) and thrombopoietin (liver)
• Mesenchymal stem cells
• are multipotent /adherent stromal cells that
can differentiate into a variety of cell types,
including osteoblasts, chondrocytes, myocytes
and adipocytes.
• Needed for stromal cell formation
• Together with osteoblasts they form
niches and provide growth factors,
adhesion molecules and cytokines
which support stem cells (protein called
jagged found on stromal cells binds to a
receptor NOTCH on stem cells which
then becomes a transcription factor
involved in the cell cycle.
• Found in peripheral blood in low
numbers
• Mobilization:
• Process necessary for
blood cells to exit the
bone marrow
• Cells cross blood vessel
endothelium
• This is enhanced by a
growth factor called
granulocyte colony-
stimulating factor (G-CSF)
• Reverse process – stem
cell homing is dependent
on stromal-derived factor
1(SDF-1)
 Tissue-specific Stem cells
• Stem cells
• Pluripotent i.e can generate various types of tissues
• Donor hamopoietic cells may contribute to tissues such as neurons, liver and
muscle
• This contributes to stem cell “plasticity”
 Regulation of Haemopoiesis
• Haemopoiesis
• Begins with stem cell division
• Stem cell plus another cell committed to differentiation
• Several transcription factors regulate survival of stem cells:
• SCL, GATA-2, NOTCH
• Transcription factors which allow for differentiation along
cell lineages:
• GATA-1 and FOG-1
 HAEMOPOIETIC GROWTH FACTORS
• HGFs
• Are glycoprotein hormones that regulate the proliferation and differentiation of
haemopoietic progenitor cells and the function of mature blood cells
• May act locally (paracrine) where produced by cell to cell contact or circulate in the
plasma (endocrine)
• Bind to ECM to form niches to which stem and progenitor cells may adhere
• Cause cell proliferation, differentiation, maturation, prevention apoptosis, affect
function of mature cells
• Share common properties
• Act at different stages of haemopoiesis (see Table 1.3; Figure 1.7)
• Important feature: 2 or more factors may synergize in stimulating a particular cell to
proliferate/differentiate
• Action of one growth factor on a cell may stimulate production of another growth
factor on another cell
General characteristics of myeloid and lymphoid growth factors
•Glycoproteins that act at very low concentrations
•Act hierarchically
•Usually produced by many cell types
•Usually affect more than one lineage
•Usually active on stem/progenitor cells and on functional end cells
•Usually show synergistic or additive interactions with other growth
factors
•Often act on the neoplastic equivalent of a normal cell
•Multiple actions: proliferation, differentiation, maturation, functional
activation, prevention of apoptosis of progenitor cells
 HAEMOPOIETIC GROWTH FACTORS
•Act on stromal cells
• IL-1
• TNF
•Act on pluripotential stem cells
• SCF
• FLT3-L
• VEGF
•Act on multipotential progenitor cells
• IL-3
• GM-CSF
• IL-6
• G-CSF
• Thrombopoietin
•Act on committed progenitor cells
• G-CSF*
• M-CSF
• IL-5 (eosinophil-CSF)
• Erythropoietin
• Thrombopoietin*
CSF, colony-stimulating factor; FLT3-L, FLT3 ligand; G-CSF, granulocyte colony-stimulating factor; GM-CSF, granulocyte–
macrophage colony-stimulating factor; IL, interleukin; M-CSF, macrophage colony-stimulating factor; SCF, stem cell factor; TNF,
tumour necrosis factor; VEGF, vascular endothelial growth factor.
•* These also act synergistically with early acting factors on pluripotential progenitors.
• Growth factors with overlapping activities:
• Interleukin-3 (IL-3); Granulocyte-macrophage colony-stimulating
factor (GM-CSF); Granulocyte colony-stimulating factor (G-CSF);
Thrombopoietin
• G-SCF and thrombopoietin
• Enhance the effects of SCF, FLT-L, IL-3 AND GM-CSF on survival
and differentiation of early haemopoietic stem cells
• These maintain a pool of haemopoietic stem and progenitor cells
• Following this, later acting erythropoietin, G-CSF, M-CSF, IL-5 and
thrombopoietin act to increase production of cells as the body
needs
• E.g. infection/inflammation can stimulate production of
granulocytes or monocytes as a result of the release of Il-1 and
tumour necrosis factor (TNF)
 GROWTH FACTOR RECEPTORS AND
SIGNAL TRANSDUCTION
• Effects of growth factors are mediated through specific receptors on target cells
• Most receptors belong the haemopoietin receptor superfamily
• Pathways activated:
• JAK/STAT:
• Janus-associated kinase
• Activates transcription of certain genes
• Mutation of JAK2 gene causes polycythemia rubra vera
• MAP KINASE
• Mitogen-activated protein kinase
• PI3
• Phosphatidylinositol 3 kinase pathway
 THE CELL CYCLE
• Interphase
• Chromosomes duplicated
• Cell growth occurs
• Divided into 3 phases
• G1
• Cell begins to replicate
• S phase
• DNA doubles and chromosomes
replicate
• G2 phase:
• Cell organelles are copied;
cytoplasmic volume restored
 THE CELL CYCLE
• Cell cycle:
• M phase
• Mitosis and cytokinesis
• Cell cycle:
• Controlled by 2 checkpoints
• Act as brakes to coordinate
division process at the end
of G1 and G2 phases
• Coordinate division process
at end of G1 and G2 phases
• Molecules which control
these checkpoints:
• Cyclin-dependent
protein kinases(Cdk)
• Phosphorylate
downstream
protein targets
• Cyclins
• Bind to Cdks and
regulate their
activity
• See video
resources
 APOPTOSIS
• Regulated process of physiological cell death in
which individual cells are triggered to activate
intracellular proteins that lead to the death of the
cell
• Characterized by:
• Cell shrinkage
• Condensation of the nuclear chromatin
• Fragmentation of the nucleus
• Cleavage of DNA at internucleosomal sites
• Important for maintaining tissue homeostasis in
haemopoiesis and lymphocyte development
• Process of Apoptosis:
• Caspases (intracellular cysteine proteases) become activated
following cleavage
• This leads to endonuclease digestion of DNA and disintegration of
the cell skeleton
• 2 pathways:
• Pathway 1:
• Signally through membrane proteins such as Fas or TNF
receptors via their intracellular death domain
• Pathway 2:
• Release of cytochrome c from mitochondria
• DNA damage induced by irradiation/chemotherapy may activate this
pathway
• Molecule that mediates apoptosis
• P53
• Protein which senses DNA damage
• Activated apoptosis by raising cell level of BAX
• BAX then increases cytochrome c release
• Also shuts down cell cycle to stop damaged cell from dividing
• Cellular level of p53 is rigidly controlled by MDM2
• Molecules that protect cells from apoptosis
• BCL-2
• Intracellular ratio of BAX and BCL-2 determines the relative
susceptibility of cells to apoptosis
• May act through regulation of cytochrome c release from
mitochondria
 ADHESION MOLECULES
• Are a family of glycoprotein molecules
• Important for the following:
• in the development and maintenance of inflammatory and immune response
• in platelet-vessel wall interactions
• Leucocyte-vessel wall interactions
• Mediate the attachment of marrow precursors, leucocytes, platelets to various
components of the extracellular matrix, to endothelium, to other surfaces and to
each other
• When found on the surface of leucocytes they are termed receptors
• They interact with ligands on the surface of potential target cells
• 3 main families:
• Immunoglobulin family
• These include receptors which react with antigens
• Selectins
• Involved mainly in leucocyte and platelet adhesion to endothelium during
inflammation and coagulation
• Integrins
• Involved in cell adhesion to ECM (collagen in wound healing; leucocyte and platelet
adhesion
• Expression may be modified:
• By extracellular and intracellular factors
• IL-1, TNF, IFN-gamma, T-cell activation, adhesion to extracellular proteins, viral
infection may upregulate the expression of adhesion molecules
• Pattern of expression of adhesion molecules
• May determine their mode of spread and tissue localization
• Adhesion molecules may also determine whether cells circulate in blood stream or remain
fixed in tissues