IDENTIFYING, TRACKING AND VISUALIZATION CML BLAST CRISIS STEM/PROGENITORS CELLS BASED ON SINGLE-CELL MASS CYTOMETRY ANALYSIS
(Abstract release date: 05/19/16)
EHA Library. Zhou H. 06/11/16; 135248; S492
Assoc. Prof. Hongsheng Zhou
Contributions
Contributions
Abstract
Abstract: S492
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:15 - 16:30
Location: Hall C11
Background
Tyrosine kinase inhibitor (TKI) has dramatically improved the outcome of CML, but it could not eliminate leukemia stem cell (LSC). Self-renewal is a key characteristic for LSC. Emerging evidence indicates that β-catenin, the Wnt pathway's central effector molecule, is required for development and maintenance of LSC in CML.
Aims
We hypothesize that the combination of β-catenin and tyrosine kinase inhibition could represent a promising strategy for preventing the development of TKI resistance as well as a novel approach in the therapy of BC CML by targeting CML-BC LSC.
Methods
Aided with single-cell mass cytometry analysis and data visualization with SPADE and viSNE, we investigated the activity of the combination of TKI with a Wnt/b-catenin signaling modulator C82 in vitro and with the C82 pro-drug PRI-724 in vivo in a xenograft murine model.
Results
Primary CML-BC cells were stained with multiple cell surface markers and intracellular survival signaling molecules and subjected to CyTOF analysis. It revealed that β-catenin expression was high in various CML stem/progenitor cells and particularly highest in CD34+CD38+CD123+Tim3+ subset. viSNE visualization demonstrated that β-catenin overexpression was associated with high levels of pCRKL, c-Myc, pAKT, pTyr, pSTAT3, and pSTAT5. Primary CML-BC cells from were then treated with C82, nilotinib, and both. viSNE visualization and SPADE tree analysis revealed that C82 and the combined regimen more significantly decreased various stem/progenitor cells, especially CD34+CD38+CD123+Tim-3+ cells, which represents for the candidate for CML-BC LSC. Next, NSG mice engrafted with CML-BCT315I cells were treated with PRI-724, nilotinib, or the combination. At the end of treatment, cells were collected from the mice and subjected to CyTOF analysis. viSNE visualization showed that PRI-724 and combination strikingly reduced CD34+b-cateninhigh, CD34+CD38-, and CD34+CD38+ different CML-BC stem/progenitor subsets. Furthermore, SPADE tree analysis revealed the combination more profoundly than PRI-724 alone reduced CD34+CD38-, CD34+CD38+ and Tim-3+GMP progenitor cells in NSG mice, which was confirmed by flow cytometry.Furthermore, PRI-724 and the combination induced CD11b expression, suggesting PRI-724 and the combination promoted CML-BC stem/progenitor cell differentiation. Notably, PRI-724 and nilotinib combination significantly inhibited CD44 in CD34+CD38+CD123high and CD34+CD38+CD123highTim-3high subsets, and Tim-3 in CD34+CD38+CD123highTim-3high subset, respectively. Finally, PRI-724 and the combination significantly improved the overall survival of CML-BC-loaded NSG mice, as compared to control and the nilotinib treatment.
Conclusion
Collectively, by identifying, tracking and visualization CML-BC LSC based on CyTOF analysis, our results show that inhibition of β-catenin effectively targets CML-BC stem/progenitor cells and prolongs survival of CML-BC-loaded NSG mice, which is further enhanced by combination with inhibition of Bcr-Abl.
Session topic: Chronic myeloid leukemia - Biology
Keyword(s): CML blast crisis, Stem and progenitor cell
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:15 - 16:30
Location: Hall C11
Background
Tyrosine kinase inhibitor (TKI) has dramatically improved the outcome of CML, but it could not eliminate leukemia stem cell (LSC). Self-renewal is a key characteristic for LSC. Emerging evidence indicates that β-catenin, the Wnt pathway's central effector molecule, is required for development and maintenance of LSC in CML.
Aims
We hypothesize that the combination of β-catenin and tyrosine kinase inhibition could represent a promising strategy for preventing the development of TKI resistance as well as a novel approach in the therapy of BC CML by targeting CML-BC LSC.
Methods
Aided with single-cell mass cytometry analysis and data visualization with SPADE and viSNE, we investigated the activity of the combination of TKI with a Wnt/b-catenin signaling modulator C82 in vitro and with the C82 pro-drug PRI-724 in vivo in a xenograft murine model.
Results
Primary CML-BC cells were stained with multiple cell surface markers and intracellular survival signaling molecules and subjected to CyTOF analysis. It revealed that β-catenin expression was high in various CML stem/progenitor cells and particularly highest in CD34+CD38+CD123+Tim3+ subset. viSNE visualization demonstrated that β-catenin overexpression was associated with high levels of pCRKL, c-Myc, pAKT, pTyr, pSTAT3, and pSTAT5. Primary CML-BC cells from were then treated with C82, nilotinib, and both. viSNE visualization and SPADE tree analysis revealed that C82 and the combined regimen more significantly decreased various stem/progenitor cells, especially CD34+CD38+CD123+Tim-3+ cells, which represents for the candidate for CML-BC LSC. Next, NSG mice engrafted with CML-BCT315I cells were treated with PRI-724, nilotinib, or the combination. At the end of treatment, cells were collected from the mice and subjected to CyTOF analysis. viSNE visualization showed that PRI-724 and combination strikingly reduced CD34+b-cateninhigh, CD34+CD38-, and CD34+CD38+ different CML-BC stem/progenitor subsets. Furthermore, SPADE tree analysis revealed the combination more profoundly than PRI-724 alone reduced CD34+CD38-, CD34+CD38+ and Tim-3+GMP progenitor cells in NSG mice, which was confirmed by flow cytometry.Furthermore, PRI-724 and the combination induced CD11b expression, suggesting PRI-724 and the combination promoted CML-BC stem/progenitor cell differentiation. Notably, PRI-724 and nilotinib combination significantly inhibited CD44 in CD34+CD38+CD123high and CD34+CD38+CD123highTim-3high subsets, and Tim-3 in CD34+CD38+CD123highTim-3high subset, respectively. Finally, PRI-724 and the combination significantly improved the overall survival of CML-BC-loaded NSG mice, as compared to control and the nilotinib treatment.
Conclusion
Collectively, by identifying, tracking and visualization CML-BC LSC based on CyTOF analysis, our results show that inhibition of β-catenin effectively targets CML-BC stem/progenitor cells and prolongs survival of CML-BC-loaded NSG mice, which is further enhanced by combination with inhibition of Bcr-Abl.
Session topic: Chronic myeloid leukemia - Biology
Keyword(s): CML blast crisis, Stem and progenitor cell
Abstract: S492
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:15 - 16:30
Location: Hall C11
Background
Tyrosine kinase inhibitor (TKI) has dramatically improved the outcome of CML, but it could not eliminate leukemia stem cell (LSC). Self-renewal is a key characteristic for LSC. Emerging evidence indicates that β-catenin, the Wnt pathway's central effector molecule, is required for development and maintenance of LSC in CML.
Aims
We hypothesize that the combination of β-catenin and tyrosine kinase inhibition could represent a promising strategy for preventing the development of TKI resistance as well as a novel approach in the therapy of BC CML by targeting CML-BC LSC.
Methods
Aided with single-cell mass cytometry analysis and data visualization with SPADE and viSNE, we investigated the activity of the combination of TKI with a Wnt/b-catenin signaling modulator C82 in vitro and with the C82 pro-drug PRI-724 in vivo in a xenograft murine model.
Results
Primary CML-BC cells were stained with multiple cell surface markers and intracellular survival signaling molecules and subjected to CyTOF analysis. It revealed that β-catenin expression was high in various CML stem/progenitor cells and particularly highest in CD34+CD38+CD123+Tim3+ subset. viSNE visualization demonstrated that β-catenin overexpression was associated with high levels of pCRKL, c-Myc, pAKT, pTyr, pSTAT3, and pSTAT5. Primary CML-BC cells from were then treated with C82, nilotinib, and both. viSNE visualization and SPADE tree analysis revealed that C82 and the combined regimen more significantly decreased various stem/progenitor cells, especially CD34+CD38+CD123+Tim-3+ cells, which represents for the candidate for CML-BC LSC. Next, NSG mice engrafted with CML-BCT315I cells were treated with PRI-724, nilotinib, or the combination. At the end of treatment, cells were collected from the mice and subjected to CyTOF analysis. viSNE visualization showed that PRI-724 and combination strikingly reduced CD34+b-cateninhigh, CD34+CD38-, and CD34+CD38+ different CML-BC stem/progenitor subsets. Furthermore, SPADE tree analysis revealed the combination more profoundly than PRI-724 alone reduced CD34+CD38-, CD34+CD38+ and Tim-3+GMP progenitor cells in NSG mice, which was confirmed by flow cytometry.Furthermore, PRI-724 and the combination induced CD11b expression, suggesting PRI-724 and the combination promoted CML-BC stem/progenitor cell differentiation. Notably, PRI-724 and nilotinib combination significantly inhibited CD44 in CD34+CD38+CD123high and CD34+CD38+CD123highTim-3high subsets, and Tim-3 in CD34+CD38+CD123highTim-3high subset, respectively. Finally, PRI-724 and the combination significantly improved the overall survival of CML-BC-loaded NSG mice, as compared to control and the nilotinib treatment.
Conclusion
Collectively, by identifying, tracking and visualization CML-BC LSC based on CyTOF analysis, our results show that inhibition of β-catenin effectively targets CML-BC stem/progenitor cells and prolongs survival of CML-BC-loaded NSG mice, which is further enhanced by combination with inhibition of Bcr-Abl.
Session topic: Chronic myeloid leukemia - Biology
Keyword(s): CML blast crisis, Stem and progenitor cell
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 16:15 - 16:30
Location: Hall C11
Background
Tyrosine kinase inhibitor (TKI) has dramatically improved the outcome of CML, but it could not eliminate leukemia stem cell (LSC). Self-renewal is a key characteristic for LSC. Emerging evidence indicates that β-catenin, the Wnt pathway's central effector molecule, is required for development and maintenance of LSC in CML.
Aims
We hypothesize that the combination of β-catenin and tyrosine kinase inhibition could represent a promising strategy for preventing the development of TKI resistance as well as a novel approach in the therapy of BC CML by targeting CML-BC LSC.
Methods
Aided with single-cell mass cytometry analysis and data visualization with SPADE and viSNE, we investigated the activity of the combination of TKI with a Wnt/b-catenin signaling modulator C82 in vitro and with the C82 pro-drug PRI-724 in vivo in a xenograft murine model.
Results
Primary CML-BC cells were stained with multiple cell surface markers and intracellular survival signaling molecules and subjected to CyTOF analysis. It revealed that β-catenin expression was high in various CML stem/progenitor cells and particularly highest in CD34+CD38+CD123+Tim3+ subset. viSNE visualization demonstrated that β-catenin overexpression was associated with high levels of pCRKL, c-Myc, pAKT, pTyr, pSTAT3, and pSTAT5. Primary CML-BC cells from were then treated with C82, nilotinib, and both. viSNE visualization and SPADE tree analysis revealed that C82 and the combined regimen more significantly decreased various stem/progenitor cells, especially CD34+CD38+CD123+Tim-3+ cells, which represents for the candidate for CML-BC LSC. Next, NSG mice engrafted with CML-BCT315I cells were treated with PRI-724, nilotinib, or the combination. At the end of treatment, cells were collected from the mice and subjected to CyTOF analysis. viSNE visualization showed that PRI-724 and combination strikingly reduced CD34+b-cateninhigh, CD34+CD38-, and CD34+CD38+ different CML-BC stem/progenitor subsets. Furthermore, SPADE tree analysis revealed the combination more profoundly than PRI-724 alone reduced CD34+CD38-, CD34+CD38+ and Tim-3+GMP progenitor cells in NSG mice, which was confirmed by flow cytometry.Furthermore, PRI-724 and the combination induced CD11b expression, suggesting PRI-724 and the combination promoted CML-BC stem/progenitor cell differentiation. Notably, PRI-724 and nilotinib combination significantly inhibited CD44 in CD34+CD38+CD123high and CD34+CD38+CD123highTim-3high subsets, and Tim-3 in CD34+CD38+CD123highTim-3high subset, respectively. Finally, PRI-724 and the combination significantly improved the overall survival of CML-BC-loaded NSG mice, as compared to control and the nilotinib treatment.
Conclusion
Collectively, by identifying, tracking and visualization CML-BC LSC based on CyTOF analysis, our results show that inhibition of β-catenin effectively targets CML-BC stem/progenitor cells and prolongs survival of CML-BC-loaded NSG mice, which is further enhanced by combination with inhibition of Bcr-Abl.
Session topic: Chronic myeloid leukemia - Biology
Keyword(s): CML blast crisis, Stem and progenitor cell
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