Involvement of CD34+ Stem Cells in Malignant Transformation in AML and MDS — Genetic Analysis of Sorted Subpopulations by Classical and Molecular Cytogenetics

  • D. Haase
  • M. Feuring-Buske
  • C. Schoch
  • C. Schäfer
  • F. Griesinger
  • C. Troff
  • B. Gahn
  • W. Hiddemann
  • B. Wörmann
Conference paper
Part of the Haematology and Blood Transfusion / Hämatologie und Bluttransfusion book series (HAEMATOLOGY, volume 39)


In this study we addressed the question whether hematopoietic stem cells characterized by the expression of CD34 are involved in the leukemogenic process in AML and MDS. For this purpose bone marrow cells were sorted according to their expression of CD34 and coexpression of CD38 or CD117 (stem cell factor receptor = CKIT). The sorted subpopulations were genetically analyzed either by cytogenetics or FISH. Successful cytogenetics of sorted CD34+ subpopulations could be performed in 24/54 pts. included in our study (AML: n = 18, MDS: n = 6). At diagnosis 18 displayed a wide variety of different clonal karyotype abnormalities in the unsorted bone marrow: add(2q); +4; 5q−; 5q− and complex anomalies; t(6;9); 6p−; −7; +8; t(8;21); −12; inv(16); i(17q); t(17;20); −21; i(21q). We analyzed sorted stem cell subpopulations with the immunophenotype CD34+/CD38± in 19 pts., in 5 pts. stem cells with the immunophenotype CD34+/ CD117± were examined. We found genetically abnormal stem cells in every subpopulation (CD34+/CD 38−, CD34+/CD38+, CD34+/CD117−, CD34+/CD117−) in every informative case (abnormal karyotype in the unsorted bone marrow and metaphases/interphase cells available for cytogenetic analysis). In 11 pts. a mosaic of normal and abnormal cells was observed in the sorted stem cell subpopulations. In five pts. secondary, progression-associated anomalies were present in the sorted stem cells. We conclude that in AML and MDS malignant transformation and disease progression occur at the level of immature hematopoietic stem cells independent from the phenotype of the leukemic bulk population and the type of genetic alteration.


Stem cells AML MDS Chromosomes Cytogenetics CD34 CD38 CD 117 


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  1. 1.
    Greaves MF (1986) Differentiation-linked leukemogenesis in lymphocytes. Science, 234, 697–704PubMedCrossRefGoogle Scholar
  2. 2.
    Fialkow PJ (1984) Clonal evolution of human myeloid leukemias. In Bishop JM, Rowley JD, Greaves M (eds) Genes and cancer. New York, Liss, p 215Google Scholar
  3. 3.
    Janssen JWG, Buschle M, Layton M, Drexler HG, Lyons J, Van den Berghe H, Heimpel H, Kubanek B, Kleihauer E, Mufti G, Bartram C, (1991) Clonal analysis of myelodysplastic syndromes: evidence of multipotent stem cell origin. Blood, 73, 248Google Scholar
  4. 4.
    Suciu S, Zeller W, Weh HJ, Hossfeld DK (1993) Immunophenotype of mitotic cells with clonal chromosome abnormalities demonstrating multilineage involvement in acute myeloid leukemia. Cancer Genet Cytogenet, 70,1PubMedCrossRefGoogle Scholar
  5. 5.
    Knuutila S, Teerenhovi L, Larramendy ML, Elonen E, Franssila KO, Nylund SJ, Timonen T, Heinonen K, Mahlamäki E, Winqvist R, and Ruutu T (1994) Cell lineage involvement of recurrent chromosomal abnormalities in hematologic neoplasms. Genes, Chromosomes and Cancer 10: 95–102.CrossRefGoogle Scholar
  6. 6.
    Dick JE, Lapidot T, Pflumio F (1991) Transplantation of normal and leukemic human bone marrow into immune-deficient mice: Development of animal models for human hematopoiesis. Immunol Rev, 124,25–43PubMedCrossRefGoogle Scholar
  7. 7.
    Krause DS, Fackler MJ, Civin CI, and May WS (1996) CD34: Structure, biology, and clinical utility. Blood, 87,1–13PubMedGoogle Scholar
  8. 8.
    Civin CI, Strauss LC, Brovall C, Fackler MJ, Schwartz JF, Shaper JH (1984) Antigenetic analysis of hematopoiesis: III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-la cells. J Immunol, 133, 157–165.PubMedGoogle Scholar
  9. 9.
    Loken MR, Shah VO, Dattilo KL, Civin CI (1987) Flow cytometric analysis of human bone marrow: II. Normal B lymphocyte development. Blood, 70,1316–1322PubMedGoogle Scholar
  10. 10.
    Huang S, Terstappen LWMM (1992) Formation of haematopoietic microenvironment and haematopoietic stem cells from single human bone marrow stem cells. Nature, 360,745–749PubMedCrossRefGoogle Scholar
  11. 11.
    Bennett J, Catovsky D, Daniel M-T, Flandrin G, Galton DAG, Gralnick HR, and Sultan C (1976) Proposals for the classification of acute leukemias. A report of the French-American-British Cooperative Group. Br J Haematol, 33, 451–458PubMedCrossRefGoogle Scholar
  12. 12.
    Greaves MF, Chan LC, Furley AJW (1986) Lineage promiscuity in hematopoietic differentiation and leukemia. Blood, 67,1–11PubMedGoogle Scholar
  13. 13.
    Terstappen LWMM, Safford M, Unterhalt M, Könemann S, Zurlutter K, Piechotka, K, Drescher M, Aul C, Büchner Th, Hiddemann W, Wörmann B. Flow (1992) Flow cytometric characterization of acute myeloid leukemia. IV. Comparison to the differentiation pathway of normal hematopoietic progenitor cells. Leukemia, 6, 993–1000PubMedGoogle Scholar
  14. 14.
    Ball ED, Lawrence D, Malnar M (1990) Correlation of CD34 and multi-drug resistance P170 with FAB and cytogenetics but not prognosis in acute myloid leukemia (AML). Blood, 76: Supp1.1, 252aGoogle Scholar
  15. 15.
    Terstappen LWMM, Loken MR (1990) Myeloid cell differentiation in normal bone marrow and acute myeloid leukemia assessed by multi-dimensional flow cytometry. Analytical Cellular Pathol, 2, 229–40Google Scholar
  16. 16.
    Lapidot T, Sirard C, Vormoor J, Murdoch B, Hoang T, Caceres-Cortes J, Minden M, Paterson B, Caligiuri MA and Dick J (1994) A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature, 367, 645–8PubMedCrossRefGoogle Scholar
  17. 17.
    Terpstra W, Prins A, Ploemacher RE, Wognum BW, Wagemaker G, Löwenberg B, and Wielenga JJ (1996) Long-term initiating capacity of a CD34- subpopulation of acute myeloid leukemia. Blood, 87,2187–2194PubMedGoogle Scholar
  18. 18.
    Reiffers J, Stoppa AM, Atta M (1993) Autologous stem cell transplantation versus chemotherapy in adult patients with acute myeloid leukemia in first remission: the BGMT group experience. Nouv Rev Fr Hemato1,35,17,1993Google Scholar
  19. 19.
    Miller AD (1992) Human gene therapy comes of age. Nature, 357, 455–60PubMedCrossRefGoogle Scholar
  20. 20.
    Wörmann B (1993) Implications of detection of minimal residual disease. Curr Opin Oncol, 5, 3–10PubMedGoogle Scholar
  21. 21.
    Brenner MK, Rill DR, Moen RC (1993) Gene-marking trace origin of relapse after autologous bone-marrow transplantation. Lancet, 341,85–6PubMedCrossRefGoogle Scholar
  22. 22.
    Heim S, Mitelman F. Acute nonlymphocytic leukemia (1989) In: Cancer cytogenetics Heim, S ed Alan R. Liss, Inc. New York, 65–110Google Scholar
  23. 23.
    Berger R, Flandrin G, Bernheim A, Le Coniat M, Vecchione D, Pacot A, Derre J, Daniel M-T, Valensi F, Sigaux F and Ochoa-Noguera ME (1987) Cytogenetic studies on 519 consecutive de novo acute nonlymphocytic leukemias. Cancer Genet Cytogenet, 29, 9–21PubMedCrossRefGoogle Scholar
  24. 24.
    Anastasi J (1993) Fluorescence in situ hybridization in leukemia. Applications in diagnosis, subclassification, and monitoring the response to therapy. Ann N Y Acad Sci, 677: 214–24PubMedCrossRefGoogle Scholar
  25. 25.
    Haase D, Feuring-Buske M, Könemann S, Fonatsch C, Troff C, Verbeek W, Pekrun A, Hiddemann W, Wörmann B (1995) Evidence for malignant transformation in acute myeloid leukemia at the level of early hematopoietic stem cells by cytogenetic analysis of CD34+ subpopulations. Blood, 86, 2906–2912PubMedGoogle Scholar
  26. 26.
    Haase D, Griesinger F, Maecker B, Feuring-Buske M, Ferrari T, Troff C, Verbeek W, Gahn B, Fonatsch C, Hiddemann W, Wörmann B (1996) Evidence for malignant transformation at the level of most immature stem cells in MDS, AML and ALL by classical cytogenetics, FISH and molecular analysis of flow-sorted CD34+ cells. Medizinische Genetik, 8, 55Google Scholar
  27. 27.
    ISCN (1985) An International System for Human Cytogenetic Nomenclature. Birth Defects: Original Article Series No.1, Vol.21, National Foundation, March of Dimes, New York.Google Scholar
  28. 28.
    Griesinger F, Ferrari T, Feuring-Buske M, Reiffers J, Hiddemann W, Wörmann B (1996) Detection of AML-1/ETO fusion transcripts in stem cell subsets and CD3+ T-lymphocytes in mobilized peripheral blood of t(8;21) AML M2 in complete remission. Blood, 88, 562a, 2238Google Scholar
  29. 29.
    Van Lom K, Hagemeijer A, Smit EME, Löwenberg B (1993) In situ hybridization on May-Grünwald Giemsa-stained bone marrow and blood smears of patients with hematologic disorders allows detection of cell lineage-specific cytogenetic abnormalities. Blood, 82, 884–888PubMedGoogle Scholar
  30. 30.
    Lim SH, Culligan D, Coutzens S, Fisher J, John S, Pollard P, Burnett A Whittacker J (1996) Molecular evidence for a common leukaemic progenitor in acute mixed lymphoid and myeloid leukaemia. Br J Haematol, 2, 31–133Google Scholar
  31. 31.
    Turhan AG, Lemoine FM, Debert C, Bonnet ML, Baillou C, Picard F, Macintyre EA, Varet B (1995) Highly purified primitive hematopoietic stem cells are PML-RARA negative and generate nonclonal progenitors in acute promyelocytic leukemia. Blood, 5,154–2161Google Scholar
  32. 32.
    Mehrothra B, George TI, Kavanau K, Avet-Loiseau H, Moore II D, Willman CL, Slovak ML, Atwater S, Head DR, Pallavicini MG (1995) Cytogenetically aberrant cells in the stem cell compartment (CD34+lin-) in acute myeloid leukemia. Blood, 6,139–1147Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • D. Haase
    • 1
  • M. Feuring-Buske
    • 1
  • C. Schoch
    • 1
  • C. Schäfer
    • 1
  • F. Griesinger
    • 1
  • C. Troff
    • 1
  • B. Gahn
    • 1
  • W. Hiddemann
    • 1
  • B. Wörmann
    • 1
  1. 1.Dept. of Hematology and OncologyGeorg-August-UniversityGöttingenGermany

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