HEMATOPOIETIC STEM CELLS

Sawang Petvises, PhD (ANU)

Department of Medical Technology, Faculty of Allied Health Sciences,

Thammasat University (Rangsit Campus)

Visiting Fellow, Stem Cell & Immunology Lab,

Research School of Biology, The Australian National University

Outline

• General concept of normal hematopoiesis

• HSC phenotypes• HSC phenotypes

• Advancement of HSC development

Hematopoietic Stem Cells (HSC)

Self-renewal

Commitment / Differentiation

Differentiated Blood Cells

Sources of HSC

• Bone marrow

• Mobilized blood• Mobilized blood

• Umbilical cord blood

All three stem cells can only make blood

Rare (1 in 15,000 Lin- cells)

+ Lin Abs Laser

Nozzle

FACS sorter

Cells separation

Ma

gn

et

HSC Isolation

+ Lin Abs Laser

Deflectionplate

+ -

Cord Blood HSC

hCD45RA

HSC ~0.0002%

Petvises S & O’Neill HC. Unpublished data

CMP

LT-HSC

MPP

CLP?

Lin-CD34+CD38-CD90+CD45RA-

Lin-CD34+CD38-CD90-CD45RA-

Lin-CD34+CD38+CD10+

Lin-CD34+CD38+IL-3RloCD45RA-

Human Hematopoiesis

GMP

MEP

RBCPlatelets Granulocytes Mac/mono B cellsT cells NK cells

EPMkP GP

Dendritic cells

Pro T Pro B Pro NKMacP

Pro DC

Lin-CD34+CD38+IL-3RloCD45RA-

Lin-CD34+CD38+IL-3R-CD45RA-

Lin-CD34+CD38+IL-3R+CD45RA-

Chao MP, Sieta J & Weissman IL, Cold Spring Harb Quan Biol. 2008

Model of Normal Hematopoiesis

G1M

DifferentiationSelf renewal

Stem cell niche

G1M

MigrationApoptosis

Upregulation of CD47

Normal HSC and Leukemic stem cells

HSCLin-CD34+CD38-CD90+

MPPLin-CD34+CD38-CD90-

AML-ETO1

Normal hematopoiesis Leukemic cellsDick JE. Ann N Y Acad Sci. 2005Jan M et al. Sci Transl Med. 2012

HSC

MPP

CMP/CLP

1 1,2 1-3

AML

1-3 1-3

1-31-3

1-31-3

1-3

1-31-3

1-31-3CMP/CLP

Progenitors/

Blast cells

Blood

CML

Blast crisis

1-31-3 1-3

1-3

Leukemic blasts

1. Initiating (block cell deaths)

2. Second event (e.g. activate CD47)

3. Third event (e.g retaining self-renewal)

Dick JE, Lapidot T. Int J Haematol. 2005, Jaiswal S et al. Cell 2009

HSC Migration

Petvises S & O’Neill HC. Immunol Cell Biol. 2011

CD47 in normal hematopoiesis and cancer

BM HSC express low level of CD47

Mobilized HSC express CD47 at high level

AML, ALL, solid tumors express high CD47

HSCMacSirp-α

CD47

AML, ALL, solid tumors express high CD47

Don’t eat me signal

Jaiswal S, Weissman IL, et al. Cell 2009

If blocked CD47 signaling

Chao MP et al. Cancer Res. 2011

Acute lymphoblastic leukemia (ALL) cells were phagocytosedby macrophages following anti-CD47 treatment.

Conclusion

• Investigation of newly HSC markers help understanding

physiology of HSC development, maintenance and

progression to malignancy

Progressive delineation of HSC phenotype advances • Progressive delineation of HSC phenotype advances

therapeutic interventions

Acknowledgements

• Faculty of Allied Health Sciences,

Thammasat University

• Prof Helen C. O’Neill,

Stem Cell & Immunology Lab, Stem Cell & Immunology Lab,

Research School of Biology,

The Australian National University

• NSTDA