BONE MARROW MESENCHYMAL STROMAL PRECURSOR CELLS FROM ACUTE MYELOID LEUKEMIA PATIENTS ARE DAMAGED BY LEUKEMIC CELLS AND THE PROCEDURE OF ALLOGENEIC BONE MARROW TRANSPLANTATION
(Abstract release date: 05/21/15)
EHA Library. Sorokina T. 06/12/15; 102689; PB1622
Dr. Tamara Sorokina
Contributions
Contributions
Abstract
Abstract: PB1622
Type: Publication Only
Background
Aims
Methods
18 newly diagnosed and 20 patients who underwent allogeneic bone marrow transplantation (alloBMT) were included in the study after informed consent. BM was aspirated prior to any treatment in the newly diagnosed group, before the conditioning and at 6 time points during 1st year after alloBMT in the corresponding group. MSC were cultured in aMEM with 10% fetal calf serum. Cumulative MSC production was counted after 3 passages. CFU-F concentration was analyzed in standard conditions. The relative expression level (REL) of different genes was measured by TaqMan RQ-PCR. As a control MSC and CFU-F from 88 healthy donors were used.
Results
The CFU-F concentration in the BM of newly diagnosed AML patients was 1/3 of the donor’s (8.7±3.7 versus 25.8±3 per 106 nucleated cells, p=0.0008). There were no correlation between CFU-F concentration and blast count in the patients BM samples. So the decrease in CFU-F concentration could not be explained by mere substitution of stromal cells by blasts, but rather it reflects the action of leukemic cells on the stromal microenvironment. In the remission BM CFU-F concentration restored (26.7±7.2 per 106 BM nucleated cells). After the alloBMT CFU-F concentration in patients’ BM decreased 3-9 fold during the next year of observation. The decrease at each time point was highly significant comparing to donors. Similar picture was observed in MSC features. Cumulative cell production in cultures of newly diagnosed AML patients was half to donor’s (5.03±0.8 x106 versus 7.6±0.8x106, p=0.07). MSC from the BM of patients before alloBMT were also slightly and insignificantly lower than from healthy donors (5.9±1 x106). In patients after alloBMT cumulative MSC production decreased 1.3-5.2 fold during the next year. The decrease at almost each time point was significant comparing to donors. Gene expression analysis revealed that in MSC of new diagnosed patients the expression of FGF2 and VEGFA significantly decreased, while REL of CSF1, FGFR1, JAG1, PPARG, PDGFRA and PDGFRB - increased. In remission REL of FGF2 and LIF were twice as high as donor’s, after alloBMT the expression of FGF2 was mainly elevated. However, REL of CSF1, PDGFB, VEGF and VCAM1 was significantly decreased at the most time points after alloBMT. It seems that expression profile of MSC at manifestation of AML reverses in the remission but still does not achieve normal levels.
Summary
During the AML development leukemic cells alter the stromal precursor cells leading to the decrease in their proliferative ability, in the level of expression of some regulatory genes and in the number of CFU-F in the BM. Chemotherapy used for induction the remission restores the stromal precursor cells incompletely. Conditioning regiments used for the alloBMT significantly damage both types of studied stromal precursors, and the effect lasted at least for 1 year. Thus, both AML cells and chemotherapeutic treatment affect BM hematopoietic microenvironment.
Keyword(s): Acute myeloid leukemia, Allogeneic bone marrow transplant, Mesenchymal stem cell, Stromal cell
Session topic: Publication Only
Type: Publication Only
Background
Leukemic cells and high dose chemotherapy affect both hematopoietic and stromal precursor cells. Changes in the hematopoiesis occurring in acute myeloid leukemia (AML) are well characterized, however, the mechanisms of alterations in stromal microenvironment during the debut of AML and pre-transplantational conditioning are still obscure.
Aims
This study aimed to analyze the alterations in the characteristics of human multipotent mesenchymal stromal cells (MSC) and their more differentiated progeny – fibroblastic colony forming units (CFU-F) derived from the bone marrow (BM) of AML patients.
Methods
18 newly diagnosed and 20 patients who underwent allogeneic bone marrow transplantation (alloBMT) were included in the study after informed consent. BM was aspirated prior to any treatment in the newly diagnosed group, before the conditioning and at 6 time points during 1st year after alloBMT in the corresponding group. MSC were cultured in aMEM with 10% fetal calf serum. Cumulative MSC production was counted after 3 passages. CFU-F concentration was analyzed in standard conditions. The relative expression level (REL) of different genes was measured by TaqMan RQ-PCR. As a control MSC and CFU-F from 88 healthy donors were used.
Results
The CFU-F concentration in the BM of newly diagnosed AML patients was 1/3 of the donor’s (8.7±3.7 versus 25.8±3 per 106 nucleated cells, p=0.0008). There were no correlation between CFU-F concentration and blast count in the patients BM samples. So the decrease in CFU-F concentration could not be explained by mere substitution of stromal cells by blasts, but rather it reflects the action of leukemic cells on the stromal microenvironment. In the remission BM CFU-F concentration restored (26.7±7.2 per 106 BM nucleated cells). After the alloBMT CFU-F concentration in patients’ BM decreased 3-9 fold during the next year of observation. The decrease at each time point was highly significant comparing to donors. Similar picture was observed in MSC features. Cumulative cell production in cultures of newly diagnosed AML patients was half to donor’s (5.03±0.8 x106 versus 7.6±0.8x106, p=0.07). MSC from the BM of patients before alloBMT were also slightly and insignificantly lower than from healthy donors (5.9±1 x106). In patients after alloBMT cumulative MSC production decreased 1.3-5.2 fold during the next year. The decrease at almost each time point was significant comparing to donors. Gene expression analysis revealed that in MSC of new diagnosed patients the expression of FGF2 and VEGFA significantly decreased, while REL of CSF1, FGFR1, JAG1, PPARG, PDGFRA and PDGFRB - increased. In remission REL of FGF2 and LIF were twice as high as donor’s, after alloBMT the expression of FGF2 was mainly elevated. However, REL of CSF1, PDGFB, VEGF and VCAM1 was significantly decreased at the most time points after alloBMT. It seems that expression profile of MSC at manifestation of AML reverses in the remission but still does not achieve normal levels.
Summary
During the AML development leukemic cells alter the stromal precursor cells leading to the decrease in their proliferative ability, in the level of expression of some regulatory genes and in the number of CFU-F in the BM. Chemotherapy used for induction the remission restores the stromal precursor cells incompletely. Conditioning regiments used for the alloBMT significantly damage both types of studied stromal precursors, and the effect lasted at least for 1 year. Thus, both AML cells and chemotherapeutic treatment affect BM hematopoietic microenvironment.
Keyword(s): Acute myeloid leukemia, Allogeneic bone marrow transplant, Mesenchymal stem cell, Stromal cell
Session topic: Publication Only
Abstract: PB1622
Type: Publication Only
Background
Aims
Methods
18 newly diagnosed and 20 patients who underwent allogeneic bone marrow transplantation (alloBMT) were included in the study after informed consent. BM was aspirated prior to any treatment in the newly diagnosed group, before the conditioning and at 6 time points during 1st year after alloBMT in the corresponding group. MSC were cultured in aMEM with 10% fetal calf serum. Cumulative MSC production was counted after 3 passages. CFU-F concentration was analyzed in standard conditions. The relative expression level (REL) of different genes was measured by TaqMan RQ-PCR. As a control MSC and CFU-F from 88 healthy donors were used.
Results
The CFU-F concentration in the BM of newly diagnosed AML patients was 1/3 of the donor’s (8.7±3.7 versus 25.8±3 per 106 nucleated cells, p=0.0008). There were no correlation between CFU-F concentration and blast count in the patients BM samples. So the decrease in CFU-F concentration could not be explained by mere substitution of stromal cells by blasts, but rather it reflects the action of leukemic cells on the stromal microenvironment. In the remission BM CFU-F concentration restored (26.7±7.2 per 106 BM nucleated cells). After the alloBMT CFU-F concentration in patients’ BM decreased 3-9 fold during the next year of observation. The decrease at each time point was highly significant comparing to donors. Similar picture was observed in MSC features. Cumulative cell production in cultures of newly diagnosed AML patients was half to donor’s (5.03±0.8 x106 versus 7.6±0.8x106, p=0.07). MSC from the BM of patients before alloBMT were also slightly and insignificantly lower than from healthy donors (5.9±1 x106). In patients after alloBMT cumulative MSC production decreased 1.3-5.2 fold during the next year. The decrease at almost each time point was significant comparing to donors. Gene expression analysis revealed that in MSC of new diagnosed patients the expression of FGF2 and VEGFA significantly decreased, while REL of CSF1, FGFR1, JAG1, PPARG, PDGFRA and PDGFRB - increased. In remission REL of FGF2 and LIF were twice as high as donor’s, after alloBMT the expression of FGF2 was mainly elevated. However, REL of CSF1, PDGFB, VEGF and VCAM1 was significantly decreased at the most time points after alloBMT. It seems that expression profile of MSC at manifestation of AML reverses in the remission but still does not achieve normal levels.
Summary
During the AML development leukemic cells alter the stromal precursor cells leading to the decrease in their proliferative ability, in the level of expression of some regulatory genes and in the number of CFU-F in the BM. Chemotherapy used for induction the remission restores the stromal precursor cells incompletely. Conditioning regiments used for the alloBMT significantly damage both types of studied stromal precursors, and the effect lasted at least for 1 year. Thus, both AML cells and chemotherapeutic treatment affect BM hematopoietic microenvironment.
Keyword(s): Acute myeloid leukemia, Allogeneic bone marrow transplant, Mesenchymal stem cell, Stromal cell
Session topic: Publication Only
Type: Publication Only
Background
Leukemic cells and high dose chemotherapy affect both hematopoietic and stromal precursor cells. Changes in the hematopoiesis occurring in acute myeloid leukemia (AML) are well characterized, however, the mechanisms of alterations in stromal microenvironment during the debut of AML and pre-transplantational conditioning are still obscure.
Aims
This study aimed to analyze the alterations in the characteristics of human multipotent mesenchymal stromal cells (MSC) and their more differentiated progeny – fibroblastic colony forming units (CFU-F) derived from the bone marrow (BM) of AML patients.
Methods
18 newly diagnosed and 20 patients who underwent allogeneic bone marrow transplantation (alloBMT) were included in the study after informed consent. BM was aspirated prior to any treatment in the newly diagnosed group, before the conditioning and at 6 time points during 1st year after alloBMT in the corresponding group. MSC were cultured in aMEM with 10% fetal calf serum. Cumulative MSC production was counted after 3 passages. CFU-F concentration was analyzed in standard conditions. The relative expression level (REL) of different genes was measured by TaqMan RQ-PCR. As a control MSC and CFU-F from 88 healthy donors were used.
Results
The CFU-F concentration in the BM of newly diagnosed AML patients was 1/3 of the donor’s (8.7±3.7 versus 25.8±3 per 106 nucleated cells, p=0.0008). There were no correlation between CFU-F concentration and blast count in the patients BM samples. So the decrease in CFU-F concentration could not be explained by mere substitution of stromal cells by blasts, but rather it reflects the action of leukemic cells on the stromal microenvironment. In the remission BM CFU-F concentration restored (26.7±7.2 per 106 BM nucleated cells). After the alloBMT CFU-F concentration in patients’ BM decreased 3-9 fold during the next year of observation. The decrease at each time point was highly significant comparing to donors. Similar picture was observed in MSC features. Cumulative cell production in cultures of newly diagnosed AML patients was half to donor’s (5.03±0.8 x106 versus 7.6±0.8x106, p=0.07). MSC from the BM of patients before alloBMT were also slightly and insignificantly lower than from healthy donors (5.9±1 x106). In patients after alloBMT cumulative MSC production decreased 1.3-5.2 fold during the next year. The decrease at almost each time point was significant comparing to donors. Gene expression analysis revealed that in MSC of new diagnosed patients the expression of FGF2 and VEGFA significantly decreased, while REL of CSF1, FGFR1, JAG1, PPARG, PDGFRA and PDGFRB - increased. In remission REL of FGF2 and LIF were twice as high as donor’s, after alloBMT the expression of FGF2 was mainly elevated. However, REL of CSF1, PDGFB, VEGF and VCAM1 was significantly decreased at the most time points after alloBMT. It seems that expression profile of MSC at manifestation of AML reverses in the remission but still does not achieve normal levels.
Summary
During the AML development leukemic cells alter the stromal precursor cells leading to the decrease in their proliferative ability, in the level of expression of some regulatory genes and in the number of CFU-F in the BM. Chemotherapy used for induction the remission restores the stromal precursor cells incompletely. Conditioning regiments used for the alloBMT significantly damage both types of studied stromal precursors, and the effect lasted at least for 1 year. Thus, both AML cells and chemotherapeutic treatment affect BM hematopoietic microenvironment.
Keyword(s): Acute myeloid leukemia, Allogeneic bone marrow transplant, Mesenchymal stem cell, Stromal cell
Session topic: Publication Only
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