HEMATOPOIETIC STEM CELLS CAN BE SEPARATED FROM LEUKEMIC CELLS IN A SUBGROUP OF ACUTE LYMPHOBLASTIC LEUKEMIA PATIENTS
(Abstract release date: 05/19/16)
EHA Library. Lutz C. 06/09/16; 132423; E874
Dr. Christoph Lutz
Contributions
Contributions
Abstract
Abstract: E874
Type: Eposter Presentation
Background
Aldehyde dehydrogenase (ALDH) activity which is involved in the metabolism of intracellular aldehydes has been demonstrated to function as a marker for stem cell activity in various tissues including the hematological system. In combination with CD34-positivity and CD38-negativity high ALDH activity (CD34+CD38-ALDH+) has been shown to be a reliable tool for the separation of hematopoietic stem cells (HSC) from leukemic cells in a subgroup of patients with acute myeloid leukemia (AML). In B-cell acute lymphoblastic leukemia (B-ALL) successful separation of normal HSC has so far been limited to a subgroup of patients. These isolations were mostly guided by CD19-negativity in combination with the CD34+CD38- phenotype. However, in some cases leukemia initiating cells were also detectable in the pool of CD19- cells indicating that better isolation strategies are necessary.
Aims
In this study we sought to investigate the value of ALDH-activity for the isolation of HSC from leukemic cells in adult B-ALL.
Methods
Between 2011 and 2015 bone marrow (BM) aspirates of 15 ALL patients were collected after written informed consent and stratified in ALDH-numerous (≥1.9% ALDH+ cells) and ALDH-rare (<1.9% ALDH+ cells) cases. The cut-off value of 1,9% was determined by the amount of ALDH+ cells in healthy bone marrow controls. HSC candidates (CD34+CD38-ALDH+) and various subpopulations were analyzed for the presence of clonal marker and functionally tested in in vitro assays.
Results
In ALDH-rare B-ALL clonal-marker negative HSC could be reliably separated by the CD34+CD38-ALDH+ phenotype, whereas this separation was not possible in ALDH-numerous B-ALL. Functional assays confirmed the HSC-potential of isolated cells and showed increased long- and short-term colony-initiating activity. Further analysis showed that in ALDH-rare ALL HSC-potential was uniformly restricted to CD19- cells. However, addition of ALDH-activity was necessary to further enrich for HSC-activity as CD34+CD38-CD19-ALDH- cells contained no colony forming potential.
Conclusion
In summary, we provide a method to separate functionally normal HSC from leukemic cells in a subgroup of B-ALL patients that can be identified by their overall low percentage of cells with high ALDH-activity. This protocol thereby allows comparative analyses of patient matched HSC and leukemic cells in order to improve the understanding of leukemic evolution in adult B-ALL.
Session topic: E-poster
Keyword(s): Aldehyde dehydrogenase, ALL, Hematopoietic stem cell, Leukemia cells
Type: Eposter Presentation
Background
Aldehyde dehydrogenase (ALDH) activity which is involved in the metabolism of intracellular aldehydes has been demonstrated to function as a marker for stem cell activity in various tissues including the hematological system. In combination with CD34-positivity and CD38-negativity high ALDH activity (CD34+CD38-ALDH+) has been shown to be a reliable tool for the separation of hematopoietic stem cells (HSC) from leukemic cells in a subgroup of patients with acute myeloid leukemia (AML). In B-cell acute lymphoblastic leukemia (B-ALL) successful separation of normal HSC has so far been limited to a subgroup of patients. These isolations were mostly guided by CD19-negativity in combination with the CD34+CD38- phenotype. However, in some cases leukemia initiating cells were also detectable in the pool of CD19- cells indicating that better isolation strategies are necessary.
Aims
In this study we sought to investigate the value of ALDH-activity for the isolation of HSC from leukemic cells in adult B-ALL.
Methods
Between 2011 and 2015 bone marrow (BM) aspirates of 15 ALL patients were collected after written informed consent and stratified in ALDH-numerous (≥1.9% ALDH+ cells) and ALDH-rare (<1.9% ALDH+ cells) cases. The cut-off value of 1,9% was determined by the amount of ALDH+ cells in healthy bone marrow controls. HSC candidates (CD34+CD38-ALDH+) and various subpopulations were analyzed for the presence of clonal marker and functionally tested in in vitro assays.
Results
In ALDH-rare B-ALL clonal-marker negative HSC could be reliably separated by the CD34+CD38-ALDH+ phenotype, whereas this separation was not possible in ALDH-numerous B-ALL. Functional assays confirmed the HSC-potential of isolated cells and showed increased long- and short-term colony-initiating activity. Further analysis showed that in ALDH-rare ALL HSC-potential was uniformly restricted to CD19- cells. However, addition of ALDH-activity was necessary to further enrich for HSC-activity as CD34+CD38-CD19-ALDH- cells contained no colony forming potential.
Conclusion
In summary, we provide a method to separate functionally normal HSC from leukemic cells in a subgroup of B-ALL patients that can be identified by their overall low percentage of cells with high ALDH-activity. This protocol thereby allows comparative analyses of patient matched HSC and leukemic cells in order to improve the understanding of leukemic evolution in adult B-ALL.
Session topic: E-poster
Keyword(s): Aldehyde dehydrogenase, ALL, Hematopoietic stem cell, Leukemia cells
Abstract: E874
Type: Eposter Presentation
Background
Aldehyde dehydrogenase (ALDH) activity which is involved in the metabolism of intracellular aldehydes has been demonstrated to function as a marker for stem cell activity in various tissues including the hematological system. In combination with CD34-positivity and CD38-negativity high ALDH activity (CD34+CD38-ALDH+) has been shown to be a reliable tool for the separation of hematopoietic stem cells (HSC) from leukemic cells in a subgroup of patients with acute myeloid leukemia (AML). In B-cell acute lymphoblastic leukemia (B-ALL) successful separation of normal HSC has so far been limited to a subgroup of patients. These isolations were mostly guided by CD19-negativity in combination with the CD34+CD38- phenotype. However, in some cases leukemia initiating cells were also detectable in the pool of CD19- cells indicating that better isolation strategies are necessary.
Aims
In this study we sought to investigate the value of ALDH-activity for the isolation of HSC from leukemic cells in adult B-ALL.
Methods
Between 2011 and 2015 bone marrow (BM) aspirates of 15 ALL patients were collected after written informed consent and stratified in ALDH-numerous (≥1.9% ALDH+ cells) and ALDH-rare (<1.9% ALDH+ cells) cases. The cut-off value of 1,9% was determined by the amount of ALDH+ cells in healthy bone marrow controls. HSC candidates (CD34+CD38-ALDH+) and various subpopulations were analyzed for the presence of clonal marker and functionally tested in in vitro assays.
Results
In ALDH-rare B-ALL clonal-marker negative HSC could be reliably separated by the CD34+CD38-ALDH+ phenotype, whereas this separation was not possible in ALDH-numerous B-ALL. Functional assays confirmed the HSC-potential of isolated cells and showed increased long- and short-term colony-initiating activity. Further analysis showed that in ALDH-rare ALL HSC-potential was uniformly restricted to CD19- cells. However, addition of ALDH-activity was necessary to further enrich for HSC-activity as CD34+CD38-CD19-ALDH- cells contained no colony forming potential.
Conclusion
In summary, we provide a method to separate functionally normal HSC from leukemic cells in a subgroup of B-ALL patients that can be identified by their overall low percentage of cells with high ALDH-activity. This protocol thereby allows comparative analyses of patient matched HSC and leukemic cells in order to improve the understanding of leukemic evolution in adult B-ALL.
Session topic: E-poster
Keyword(s): Aldehyde dehydrogenase, ALL, Hematopoietic stem cell, Leukemia cells
Type: Eposter Presentation
Background
Aldehyde dehydrogenase (ALDH) activity which is involved in the metabolism of intracellular aldehydes has been demonstrated to function as a marker for stem cell activity in various tissues including the hematological system. In combination with CD34-positivity and CD38-negativity high ALDH activity (CD34+CD38-ALDH+) has been shown to be a reliable tool for the separation of hematopoietic stem cells (HSC) from leukemic cells in a subgroup of patients with acute myeloid leukemia (AML). In B-cell acute lymphoblastic leukemia (B-ALL) successful separation of normal HSC has so far been limited to a subgroup of patients. These isolations were mostly guided by CD19-negativity in combination with the CD34+CD38- phenotype. However, in some cases leukemia initiating cells were also detectable in the pool of CD19- cells indicating that better isolation strategies are necessary.
Aims
In this study we sought to investigate the value of ALDH-activity for the isolation of HSC from leukemic cells in adult B-ALL.
Methods
Between 2011 and 2015 bone marrow (BM) aspirates of 15 ALL patients were collected after written informed consent and stratified in ALDH-numerous (≥1.9% ALDH+ cells) and ALDH-rare (<1.9% ALDH+ cells) cases. The cut-off value of 1,9% was determined by the amount of ALDH+ cells in healthy bone marrow controls. HSC candidates (CD34+CD38-ALDH+) and various subpopulations were analyzed for the presence of clonal marker and functionally tested in in vitro assays.
Results
In ALDH-rare B-ALL clonal-marker negative HSC could be reliably separated by the CD34+CD38-ALDH+ phenotype, whereas this separation was not possible in ALDH-numerous B-ALL. Functional assays confirmed the HSC-potential of isolated cells and showed increased long- and short-term colony-initiating activity. Further analysis showed that in ALDH-rare ALL HSC-potential was uniformly restricted to CD19- cells. However, addition of ALDH-activity was necessary to further enrich for HSC-activity as CD34+CD38-CD19-ALDH- cells contained no colony forming potential.
Conclusion
In summary, we provide a method to separate functionally normal HSC from leukemic cells in a subgroup of B-ALL patients that can be identified by their overall low percentage of cells with high ALDH-activity. This protocol thereby allows comparative analyses of patient matched HSC and leukemic cells in order to improve the understanding of leukemic evolution in adult B-ALL.
Session topic: E-poster
Keyword(s): Aldehyde dehydrogenase, ALL, Hematopoietic stem cell, Leukemia cells
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