SpringerProtocols: Abstract: Identification of Murine and Human Acute Myeloid Leukemia Stem Cells

3. Identification of Murine and Human Acute Myeloid Leukemia Stem Cells

By: Aniruddha J. Deshpande², Farid Ahmed², Christian Buske²

Abstract

Full Text

Download PDF (674K)

There is now compelling evidence to show that tumors, once believed to be a homogeneous mass of abnormally proliferative cells, comprise a heterogeneous population of transformed cells resembling the hierarchically organized populations in the corresponding tissue. At the top of this tumor hierarchy are the cancer stem cells (CSCs) which are critical for tumor growth and maintenance as they constantly replenish the tumor bulk and are believed to be resistant to most conventional chemotherapies. The first evidence for both malignant hierarchy and CSCs came from studies on acute myeloid leukemia (AML) (1, 2) followed by mounting evidence in other types of cancer ( 3–5 ) . Murine models of leukemia have been the proving ground for the elucidation of key novel concepts in this nascent field of CSC research. A spate of recent studies highlighting the importance of CSCs in cancer has accentuated the need for identifying and characterizing these cells. The frequency of CSCs in the leukemic bulk (LSCs) is usually estimated by in vivo limiting-dilution transplantation assays of leukemic cells into recipient animal hosts in which identical leukemias can be regenerated. Each cell that is capable of propagating the leukemia in secondary recipients is termed a leukemia-propagating cell (LPC).

LSC candidates have been typically identified by the systematic dissection of compartments within a heterogeneous tumor to determine the subpopulation which shows maximal enrichment for LPCs. This is performed by determining the frequency of LPCs in each of those purified subpopulations, as has been described in both human and murine models of leukemia ( 1, 2, 6 ). It is important to mention that even though leukemia repopulation is often used as a surrogate for LSC estimation, the term LPCs can be equated with LSCs only if the phenotypic heterogeneity of the original tumor mass is regenerated upon tumor propagation into secondary recipients.

The first part of the chapter deals with the estimation of LPCs in syngenic or congenic murine–murine models. In the second part, murine xenograft models for the estimation of LSCs in human leukemias are described.

Affiliation(s): (2) GSF National Research Center for Environment and Health and the Clinical Cooperative Group – Leukemia, Department of Medicine III, Klinikum Grosshadern, Munich, Germany

Book Title: Cancer Stem Cells: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 568 | Pub. Date: Mar-01-2008 | Page Range: 21-35 | DOI: 10.1007/978-1-59745-280-9_3

Subject: Cancer Research

Key Words: AML - Cancer stem cells - Leukemia - Limiting dilution assays - Leukemic stem cells - Mouse bone marrow transplantation

References

Download References

1.	Bonnet, D. and J.E. Dick (1997), Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med. 3(7): p. 730–7.

2.	Lapidot, T., et al. (1994), A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature. 367(6464): p. 645–8.

3.	Al-Hajj, M., et al. (2003), Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A. 100(7): p. 3983–8.

4.	Singh, S.K., et al. (2004), Identification of human brain tumour initiating cells. Nature. 432(7015): 396–401.

5.	O’Brien, C.A., et al. (2007), A human colon cancer cell capable of initiating tumour growth in immuno-deficient mice. Nature. 445(7123): p. 106–10.

6.	Deshpande, A.J., et al. (2006), Acute myeloid leukemia is propagated by a leukemic stem cell with lymphoid characteristics in a mouse model of CALM/AF10-positive leukemia. Cancer Cell. 10(5): p. 363–74.

7.	Szilvassy, S.J., et al. (1990), Quantitative assay for totipotent reconstituting hematopoietic stem cells by a competitive repopulation strategy. Proc Natl Acad Sci U S A. 87(22): p. 8736–40.

8.	Cozzio, A., et al. (2003), Similar MLL-associated leukemias arising from self-renewing stem cells and short-lived myeloid progenitors. Genes Dev. 17(24): p. 3029–35.

9.	So, C.W., et al. (2003), MLL-GAS7 transforms multipotent hematopoietic progenitors and induces mixed lineage leukemias in mice. Cancer Cell. 3(2): p. 161–71.

10.	Huntly, B.J., et al. (2004), MOZ-TIF2, but not BCR-ABL, confers properties of leukemic stem cells to committed murine hematopoietic progenitors. Cancer Cell. 6(6): p. 587–96.

11.	Ailles, L.E., B. Gerhard, and D.E. Hogge (1997), Detection and characterization of primitive malignant and normal progenitors in patients with acute myelogenous leukemia using long-term coculture with supportive feeder layers and cytokines. Blood. 90(7): p. 2555–64.

12.	Buick, R.N., J.E. Till, and E.A. McCulloch (1977), Colony assay for proliferative blast cells circulating in myeloblastic leukaemia. Lancet. 1(8016): p. 862–3.

13.	Sutherland, H.J., et al. (1991), Differential regulation of primitive human hematopoietic cells in long-term cultures maintained on genetically engineered murine stromal cells. Blood. 78(3): p. 666–72.

14.	Ailles, L.E., et al. (1999), Growth characteristics of acute myelogenous leukemia progenitors that initiate malignant hematopoiesis in nonobese diabetic/severe combined immunodeficient mice. Blood. 94(5): p. 1761–72.

15.	Pearce, D.J., et al. (2006), AML engraftment in the NOD/SCID assay reflects the outcome of AML: implications for our understanding of the heterogeneity of AML. Blood. 107(3): p. 1166–73.

16.	Feuring-Buske, M., et al. (2002), A diphtheria toxin-interleukin 3 fusion protein is cytotoxic to primitive acute myeloid leukemia progenitors but spares normal progenitors. Cancer Res. 62(6): p. 1730–6.

17.	Terpstra, W., et al. (1997), Diphtheria toxin fused to granulocyte-macrophage colony-stimulating factor eliminates acute myeloid leukemia cells with the potential to initiate leukemia in immunodeficient mice, but spares normal hemopoietic stem cells. Blood. 90(9): p. 3735–42.

18.	Hosen, N., et al. (2007), CD96 is a leukemic stem cell-specific marker in human acute myeloid leukemia. Proc Natl Acad Sci U S A. 104(26): p. 11008–13.

Comments (Loading...)

Post comment/View all comments