DYSFUNCTION OF BONE MARROW ENDOTHELIAL PROGENITOR CELLS FROM SUBJECTS WITH POOR GRAFT FUNCTION FOLLOWING ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION CAN BE IMPROVED BY ATORVASTATIN
(Abstract release date: 05/19/16)
EHA Library. Shi M. 06/09/16; 132681; E1132
Mrs. Minmin Shi
Contributions
Contributions
Abstract
Abstract: E1132
Type: Eposter Presentation
Background
Poor graft function （PGF) is a serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the mechanisms are poorly understood. Murine studies suggest that endothelial progenitor cells (EPCs) are preferential supporting cells for hematopoietic stem cells (HSCs) in the bone marrow (BM) microenvironment. Our previous prospective nested case-control study suggested that a lower frequency of EPCs, which represent a key cellular component of the BM microenvironment, was an independent risk factor for the occurrence of poor graft function after allo-HSCT. However, the functional role of bone marrow EPCs in the subjects with poor graft function has never been reported. Moreover, approaches for improving the dysfunction of bone marrow EPCs in subjects with PGF are lacking. Atorvastatin has been reported to improve the mobilization and function of circulating EPCs in a number of diseases, such as heart disease, diabetes, ischemic stroke and others. Nevertheless, no previous studies have focused on the roles of atorvastatin on bone marrow-derived EPCs in subjects with poor graft function following allo-HSCT.
Aims
In this study, we evaluated the function of bone marrow EPCs in subjects with poor graft function post-allotransplant. Moreover, we investigated whether atorvastatin could enhance the number and function of bone marrow EPCs derived from subjects with poor graft function in vitro.
Methods
Three cohorts were included: subjects with poor graft function (N=15), subjects with good graft function (N=15), defined as persistent successful engraftment after allotransplant, and transplant donors as normal controls (N=15). Atorvastatin (0.5nM,5nM,500nM) was administrated to the bone marrow EPCs in subjects with poor graft function cultured for 7 days. The number and functions ofCD34(+)/CD133(+)/KDR(+)EPCs were evaluated by flow cytometry, cell counting, DiI-Ac-LDL and FITC-lectin-UEA-1 double staining，migration, tube formation test and apoptosis. Reactive Oxygen Species (ROS) level was evaluated by flow cytometry and DCFH-DA staining. Cell proliferation was determined by cell counting kit-8 assay. Protein expression for p-p38,p38, p-akt, akt was measured by flow cytometry and western blots.
Results
Dysfunctional bone marrow EPCs, which were characterized by decreased migration and angiogenesis and higher levels of reactive oxygen species and apoptosis, were found in subjects with poor graft function following allo-HSCT. The phospho-p38 expression was significantly elevated in subject with poor graft function but all groups showed similar intracellular levels of phospho-Akt. Furthermore, bone morrow EPCs derived from subjects with poor graft function were enhanced both quantitatively and functionally by 500nM atorvastatin treatment in culture on day 7 through down-regulation of the p38 MAPK pathway.
Conclusion
In summary, the current study is, to our knowledge, the first to demonstrate the dysfunctions of bone marrow EPCs in the bone marrow microenvironment of subjects with poor graft function following allo-HSCT. Moreover, atorvastatin treatment in vitro quantitatively and functionally improved the bone morrow EPCs derived from subjects with poor graft function through down-regulation of the p38 MAPK pathway. Our results indicate that atorvastatin represents a promising therapeutic approach for repairing the impaired EPCs in subjects with poor graft function post-allotransplant.
Session topic: E-poster
Keyword(s): Endothelial progenitor cell
Type: Eposter Presentation
Background
Poor graft function （PGF) is a serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the mechanisms are poorly understood. Murine studies suggest that endothelial progenitor cells (EPCs) are preferential supporting cells for hematopoietic stem cells (HSCs) in the bone marrow (BM) microenvironment. Our previous prospective nested case-control study suggested that a lower frequency of EPCs, which represent a key cellular component of the BM microenvironment, was an independent risk factor for the occurrence of poor graft function after allo-HSCT. However, the functional role of bone marrow EPCs in the subjects with poor graft function has never been reported. Moreover, approaches for improving the dysfunction of bone marrow EPCs in subjects with PGF are lacking. Atorvastatin has been reported to improve the mobilization and function of circulating EPCs in a number of diseases, such as heart disease, diabetes, ischemic stroke and others. Nevertheless, no previous studies have focused on the roles of atorvastatin on bone marrow-derived EPCs in subjects with poor graft function following allo-HSCT.
Aims
In this study, we evaluated the function of bone marrow EPCs in subjects with poor graft function post-allotransplant. Moreover, we investigated whether atorvastatin could enhance the number and function of bone marrow EPCs derived from subjects with poor graft function in vitro.
Methods
Three cohorts were included: subjects with poor graft function (N=15), subjects with good graft function (N=15), defined as persistent successful engraftment after allotransplant, and transplant donors as normal controls (N=15). Atorvastatin (0.5nM,5nM,500nM) was administrated to the bone marrow EPCs in subjects with poor graft function cultured for 7 days. The number and functions ofCD34(+)/CD133(+)/KDR(+)EPCs were evaluated by flow cytometry, cell counting, DiI-Ac-LDL and FITC-lectin-UEA-1 double staining，migration, tube formation test and apoptosis. Reactive Oxygen Species (ROS) level was evaluated by flow cytometry and DCFH-DA staining. Cell proliferation was determined by cell counting kit-8 assay. Protein expression for p-p38,p38, p-akt, akt was measured by flow cytometry and western blots.
Results
Dysfunctional bone marrow EPCs, which were characterized by decreased migration and angiogenesis and higher levels of reactive oxygen species and apoptosis, were found in subjects with poor graft function following allo-HSCT. The phospho-p38 expression was significantly elevated in subject with poor graft function but all groups showed similar intracellular levels of phospho-Akt. Furthermore, bone morrow EPCs derived from subjects with poor graft function were enhanced both quantitatively and functionally by 500nM atorvastatin treatment in culture on day 7 through down-regulation of the p38 MAPK pathway.
Conclusion
In summary, the current study is, to our knowledge, the first to demonstrate the dysfunctions of bone marrow EPCs in the bone marrow microenvironment of subjects with poor graft function following allo-HSCT. Moreover, atorvastatin treatment in vitro quantitatively and functionally improved the bone morrow EPCs derived from subjects with poor graft function through down-regulation of the p38 MAPK pathway. Our results indicate that atorvastatin represents a promising therapeutic approach for repairing the impaired EPCs in subjects with poor graft function post-allotransplant.
Session topic: E-poster
Keyword(s): Endothelial progenitor cell
Abstract: E1132
Type: Eposter Presentation
Background
Poor graft function （PGF) is a serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the mechanisms are poorly understood. Murine studies suggest that endothelial progenitor cells (EPCs) are preferential supporting cells for hematopoietic stem cells (HSCs) in the bone marrow (BM) microenvironment. Our previous prospective nested case-control study suggested that a lower frequency of EPCs, which represent a key cellular component of the BM microenvironment, was an independent risk factor for the occurrence of poor graft function after allo-HSCT. However, the functional role of bone marrow EPCs in the subjects with poor graft function has never been reported. Moreover, approaches for improving the dysfunction of bone marrow EPCs in subjects with PGF are lacking. Atorvastatin has been reported to improve the mobilization and function of circulating EPCs in a number of diseases, such as heart disease, diabetes, ischemic stroke and others. Nevertheless, no previous studies have focused on the roles of atorvastatin on bone marrow-derived EPCs in subjects with poor graft function following allo-HSCT.
Aims
In this study, we evaluated the function of bone marrow EPCs in subjects with poor graft function post-allotransplant. Moreover, we investigated whether atorvastatin could enhance the number and function of bone marrow EPCs derived from subjects with poor graft function in vitro.
Methods
Three cohorts were included: subjects with poor graft function (N=15), subjects with good graft function (N=15), defined as persistent successful engraftment after allotransplant, and transplant donors as normal controls (N=15). Atorvastatin (0.5nM,5nM,500nM) was administrated to the bone marrow EPCs in subjects with poor graft function cultured for 7 days. The number and functions ofCD34(+)/CD133(+)/KDR(+)EPCs were evaluated by flow cytometry, cell counting, DiI-Ac-LDL and FITC-lectin-UEA-1 double staining，migration, tube formation test and apoptosis. Reactive Oxygen Species (ROS) level was evaluated by flow cytometry and DCFH-DA staining. Cell proliferation was determined by cell counting kit-8 assay. Protein expression for p-p38,p38, p-akt, akt was measured by flow cytometry and western blots.
Results
Dysfunctional bone marrow EPCs, which were characterized by decreased migration and angiogenesis and higher levels of reactive oxygen species and apoptosis, were found in subjects with poor graft function following allo-HSCT. The phospho-p38 expression was significantly elevated in subject with poor graft function but all groups showed similar intracellular levels of phospho-Akt. Furthermore, bone morrow EPCs derived from subjects with poor graft function were enhanced both quantitatively and functionally by 500nM atorvastatin treatment in culture on day 7 through down-regulation of the p38 MAPK pathway.
Conclusion
In summary, the current study is, to our knowledge, the first to demonstrate the dysfunctions of bone marrow EPCs in the bone marrow microenvironment of subjects with poor graft function following allo-HSCT. Moreover, atorvastatin treatment in vitro quantitatively and functionally improved the bone morrow EPCs derived from subjects with poor graft function through down-regulation of the p38 MAPK pathway. Our results indicate that atorvastatin represents a promising therapeutic approach for repairing the impaired EPCs in subjects with poor graft function post-allotransplant.
Session topic: E-poster
Keyword(s): Endothelial progenitor cell
Type: Eposter Presentation
Background
Poor graft function （PGF) is a serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the mechanisms are poorly understood. Murine studies suggest that endothelial progenitor cells (EPCs) are preferential supporting cells for hematopoietic stem cells (HSCs) in the bone marrow (BM) microenvironment. Our previous prospective nested case-control study suggested that a lower frequency of EPCs, which represent a key cellular component of the BM microenvironment, was an independent risk factor for the occurrence of poor graft function after allo-HSCT. However, the functional role of bone marrow EPCs in the subjects with poor graft function has never been reported. Moreover, approaches for improving the dysfunction of bone marrow EPCs in subjects with PGF are lacking. Atorvastatin has been reported to improve the mobilization and function of circulating EPCs in a number of diseases, such as heart disease, diabetes, ischemic stroke and others. Nevertheless, no previous studies have focused on the roles of atorvastatin on bone marrow-derived EPCs in subjects with poor graft function following allo-HSCT.
Aims
In this study, we evaluated the function of bone marrow EPCs in subjects with poor graft function post-allotransplant. Moreover, we investigated whether atorvastatin could enhance the number and function of bone marrow EPCs derived from subjects with poor graft function in vitro.
Methods
Three cohorts were included: subjects with poor graft function (N=15), subjects with good graft function (N=15), defined as persistent successful engraftment after allotransplant, and transplant donors as normal controls (N=15). Atorvastatin (0.5nM,5nM,500nM) was administrated to the bone marrow EPCs in subjects with poor graft function cultured for 7 days. The number and functions ofCD34(+)/CD133(+)/KDR(+)EPCs were evaluated by flow cytometry, cell counting, DiI-Ac-LDL and FITC-lectin-UEA-1 double staining，migration, tube formation test and apoptosis. Reactive Oxygen Species (ROS) level was evaluated by flow cytometry and DCFH-DA staining. Cell proliferation was determined by cell counting kit-8 assay. Protein expression for p-p38,p38, p-akt, akt was measured by flow cytometry and western blots.
Results
Dysfunctional bone marrow EPCs, which were characterized by decreased migration and angiogenesis and higher levels of reactive oxygen species and apoptosis, were found in subjects with poor graft function following allo-HSCT. The phospho-p38 expression was significantly elevated in subject with poor graft function but all groups showed similar intracellular levels of phospho-Akt. Furthermore, bone morrow EPCs derived from subjects with poor graft function were enhanced both quantitatively and functionally by 500nM atorvastatin treatment in culture on day 7 through down-regulation of the p38 MAPK pathway.
Conclusion
In summary, the current study is, to our knowledge, the first to demonstrate the dysfunctions of bone marrow EPCs in the bone marrow microenvironment of subjects with poor graft function following allo-HSCT. Moreover, atorvastatin treatment in vitro quantitatively and functionally improved the bone morrow EPCs derived from subjects with poor graft function through down-regulation of the p38 MAPK pathway. Our results indicate that atorvastatin represents a promising therapeutic approach for repairing the impaired EPCs in subjects with poor graft function post-allotransplant.
Session topic: E-poster
Keyword(s): Endothelial progenitor cell
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