Normal Hematopoietic Progenitor Subsets Have Distinct Reactive Oxygen Species, BCL2 and Cell-Cycle Profiles That Are Decoupled from Maturation in Acute Myeloid Leukemia.
In acute myeloid leukemia (AML) quiescence and low oxidative state, linked to BCL2 mitochondrial regulation, endow leukemic stem cells (LSC) with treatment-resistance. LSC in CD34+ and more mature CD34- AML have heterogeneous immunophenotypes overlapping with normal stem/progenitor cells (SPC) but may be differentiated by functional markers. We therefore investigated the oxidative/reactive oxygen species (ROS) profile, its relationship with cell-cycle/BCL2 for normal SPC, and whether altered in AML and myelodysplasia (MDS). In control BM (n = 24), ROS levels were highest in granulocyte-macrophage progenitors (GMP) and CD34- myeloid precursors but megakaryocyte-erythroid progenitors had equivalent levels to CD34+CD38low immature-SPC although they were ki67high. BCL2 upregulation was specific to GMPs. This profile was also observed for CD34+SPC in MDS-without-excess-blasts (MDS-noEB, n = 12). Erythroid CD34- precursors were, however, abnormally ROS-high in MDS-noEB, potentially linking oxidative stress to cell loss. In pre-treatment AML (n = 93) and MDS-with-excess-blasts (MDS-RAEB) (n = 14), immunophenotypic mature-SPC had similar ROS levels to co-existing immature-SPC. However ROS levels varied between AMLs; Flt3ITD+/NPM1wild-type CD34+SPC had higher ROS than NPM1mutated CD34+ or CD34- SPC. An aberrant ki67lowBCL2high immunophenotype was observed in CD34+AML (most prominent in Flt3ITD AMLs) but also in CD34- AMLs and MDS-RAEB, suggesting a shared redox/pro-survival adaptation. Some patients had BCL2 overexpression in CD34+ ROS-high as well as ROS-low fractions which may be indicative of poor early response to standard chemotherapy. Thus normal SPC subsets have distinct ROS, cell-cycle, BCL2 profiles that in AML /MDS-RAEB are decoupled from maturation. The combined profile of these functional properties in AML subpopulations may be relevant to differential treatment resistance.
Department of Clinical Immunology, Institute of Immunology and Immunotherapy, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
Naeem
http://orcid.org/0000-0002-1433-8454
N
Khan
Department of Haematology, Cardiff University School of Medicine, University Hospital Wales, Cardiff, United Kingdom.
Robert K
RK
Hills
Department of Haematology, Cardiff University School of Medicine, University Hospital Wales, Cardiff, United Kingdom.
Steve
S
Knapper
Department of Clinical Immunology, Institute of Immunology and Immunotherapy, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
Lora
L
Steadman
Department of Clinical Immunology, Institute of Immunology and Immunotherapy, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
Ushna
U
Qureshi
Department of Clinical Immunology, Institute of Immunology and Immunotherapy, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
Jerrald L
JL
Rector
School of Cellular and Molecular Medicine, University of Bristol, Bristol, United Kingdom.
Charlotte
C
Bradbury
Centre for Clinical Haematology, Nottingham University Hospital NHS Trust, Nottingham, United Kingdom.
Nigel H
NH
Russell
Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom.
Paresh
P
Vyas
Department of Haematology, Cardiff University School of Medicine, University Hospital Wales, Cardiff, United Kingdom.
Alan K
AK
Burnett
Department of Medical and Molecular Genetics, King's College London School of Medicine, Guy's & St. Thomas' NHS Foundation Trust, London, United Kingdom.
David
D
Grimwade
Department of Haematology, Cardiff University School of Medicine, University Hospital Wales, Cardiff, United Kingdom.
Paul S
PS
Hole
Department of Clinical Immunology, Institute of Immunology and Immunotherapy, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
Sylvie D
SD
Freeman
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