AML ORCHESTRATES MITOCHONDRIAL METABOLISM IN BONE MARROW MESENCHYMAL STROMAL CELLS THROUGH AN INCREASE OF PGC-1Α.
(Abstract release date: 05/19/16)
EHA Library. Marlein C. 06/09/16; 132431; E882
Mr. Christopher Marlein
Contributions
Contributions
Abstract
Abstract: E882
Type: Eposter Presentation
Background
Acute Myeloid Leukaemia (AML) is biologically a heterogeneous disease which results from defects in haematopoiesis, characterised by the accumulation of haematopoietic myeloid cells in the bone marrow. Although there have been advances in the treatment of the disease, AML remains incurable for at least 80% of patients. It has been shown that AML cells have an increased mitochondrial mass and higher levels of oxidative phosphorylation (OXPHOS) compared to normal CD34+ cells, suggesting a dependency of OXPHOS for AML survival. Little is known about the mitochondrial activity of bone marrow mesenchymal stromal cells (BM-MSC), which provide a supportive environment for AML cells in the bone marrow.
Aims
Therefore the aim of this study is to determine if there is an increase in mitochondrial respiration in BM-MSCs from AML patients (AML BM-MSCs) compared to non-AML patients (normal BM-MSCs) and to understand how the difference is regulated.
Methods
Primary AML blasts were obtained from patient bone marrow. Primary AML and normal BM-MSCs were isolated from patients bone marrow, with informed consent and under approval from the UK National Research Ethics Service (LRCEref07/H0310/146), using adherence. BM-MSC were characterised using flow cytometry for expression of CD90+, CD73+, CD105+ and CD45-. Mitochondrial respiration and glycolysis rates were measured using the Seahorse XF24 extracellular flux analyser. RT-qPCR and western blotting were used to determine peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) expression and regulation. Lentiviral mediated knockdown was also used.
Results
Our results show that AML BM-MSCs have an increased ATP production after 3 days compared to normal BM-MSCs. AML BM-MSCs also show an increased oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) compared to normal BM-MSCs. This demonstrates not only an increase in mitochondrial respiration but also increased glycolysis, which correlates with the increased ATP production of the AML BM-MSCs compared to normal BM-MSCs. Mitochondrial respiration and glycolysis increase in AML BM-MSCs after co-culture with primary AML cells. This suggests that exposure to AML cells cause the increased mitochondrial respiration and glycolysis observed in the AML BM-MSCs. There is differential expression of PGC-1α mRNA and protein in AML BM-MSCs after co-culture with AML cells, suggesting regulation of mitochondrial respiration through PGC-1α.
Conclusion
These results provide evidence of an increased mitochondrial respiration and glycolysis in the AML BM-MSCs compared to the normal BM-MSCs and that the difference is orchestrated by the AML cells. In addition this difference is regulated through PGC-1α, the master regulator of mitochondrial biogenesis. We hypothesise that therapeutic intervention targeting mitochondrial activity in the AML micro-environment would make AML more susceptible to current treatment regimes.
Session topic: E-poster
Keyword(s): Acute myeloid leukemia, Mitochondria
Type: Eposter Presentation
Background
Acute Myeloid Leukaemia (AML) is biologically a heterogeneous disease which results from defects in haematopoiesis, characterised by the accumulation of haematopoietic myeloid cells in the bone marrow. Although there have been advances in the treatment of the disease, AML remains incurable for at least 80% of patients. It has been shown that AML cells have an increased mitochondrial mass and higher levels of oxidative phosphorylation (OXPHOS) compared to normal CD34+ cells, suggesting a dependency of OXPHOS for AML survival. Little is known about the mitochondrial activity of bone marrow mesenchymal stromal cells (BM-MSC), which provide a supportive environment for AML cells in the bone marrow.
Aims
Therefore the aim of this study is to determine if there is an increase in mitochondrial respiration in BM-MSCs from AML patients (AML BM-MSCs) compared to non-AML patients (normal BM-MSCs) and to understand how the difference is regulated.
Methods
Primary AML blasts were obtained from patient bone marrow. Primary AML and normal BM-MSCs were isolated from patients bone marrow, with informed consent and under approval from the UK National Research Ethics Service (LRCEref07/H0310/146), using adherence. BM-MSC were characterised using flow cytometry for expression of CD90+, CD73+, CD105+ and CD45-. Mitochondrial respiration and glycolysis rates were measured using the Seahorse XF24 extracellular flux analyser. RT-qPCR and western blotting were used to determine peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) expression and regulation. Lentiviral mediated knockdown was also used.
Results
Our results show that AML BM-MSCs have an increased ATP production after 3 days compared to normal BM-MSCs. AML BM-MSCs also show an increased oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) compared to normal BM-MSCs. This demonstrates not only an increase in mitochondrial respiration but also increased glycolysis, which correlates with the increased ATP production of the AML BM-MSCs compared to normal BM-MSCs. Mitochondrial respiration and glycolysis increase in AML BM-MSCs after co-culture with primary AML cells. This suggests that exposure to AML cells cause the increased mitochondrial respiration and glycolysis observed in the AML BM-MSCs. There is differential expression of PGC-1α mRNA and protein in AML BM-MSCs after co-culture with AML cells, suggesting regulation of mitochondrial respiration through PGC-1α.
Conclusion
These results provide evidence of an increased mitochondrial respiration and glycolysis in the AML BM-MSCs compared to the normal BM-MSCs and that the difference is orchestrated by the AML cells. In addition this difference is regulated through PGC-1α, the master regulator of mitochondrial biogenesis. We hypothesise that therapeutic intervention targeting mitochondrial activity in the AML micro-environment would make AML more susceptible to current treatment regimes.
Session topic: E-poster
Keyword(s): Acute myeloid leukemia, Mitochondria
Abstract: E882
Type: Eposter Presentation
Background
Acute Myeloid Leukaemia (AML) is biologically a heterogeneous disease which results from defects in haematopoiesis, characterised by the accumulation of haematopoietic myeloid cells in the bone marrow. Although there have been advances in the treatment of the disease, AML remains incurable for at least 80% of patients. It has been shown that AML cells have an increased mitochondrial mass and higher levels of oxidative phosphorylation (OXPHOS) compared to normal CD34+ cells, suggesting a dependency of OXPHOS for AML survival. Little is known about the mitochondrial activity of bone marrow mesenchymal stromal cells (BM-MSC), which provide a supportive environment for AML cells in the bone marrow.
Aims
Therefore the aim of this study is to determine if there is an increase in mitochondrial respiration in BM-MSCs from AML patients (AML BM-MSCs) compared to non-AML patients (normal BM-MSCs) and to understand how the difference is regulated.
Methods
Primary AML blasts were obtained from patient bone marrow. Primary AML and normal BM-MSCs were isolated from patients bone marrow, with informed consent and under approval from the UK National Research Ethics Service (LRCEref07/H0310/146), using adherence. BM-MSC were characterised using flow cytometry for expression of CD90+, CD73+, CD105+ and CD45-. Mitochondrial respiration and glycolysis rates were measured using the Seahorse XF24 extracellular flux analyser. RT-qPCR and western blotting were used to determine peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) expression and regulation. Lentiviral mediated knockdown was also used.
Results
Our results show that AML BM-MSCs have an increased ATP production after 3 days compared to normal BM-MSCs. AML BM-MSCs also show an increased oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) compared to normal BM-MSCs. This demonstrates not only an increase in mitochondrial respiration but also increased glycolysis, which correlates with the increased ATP production of the AML BM-MSCs compared to normal BM-MSCs. Mitochondrial respiration and glycolysis increase in AML BM-MSCs after co-culture with primary AML cells. This suggests that exposure to AML cells cause the increased mitochondrial respiration and glycolysis observed in the AML BM-MSCs. There is differential expression of PGC-1α mRNA and protein in AML BM-MSCs after co-culture with AML cells, suggesting regulation of mitochondrial respiration through PGC-1α.
Conclusion
These results provide evidence of an increased mitochondrial respiration and glycolysis in the AML BM-MSCs compared to the normal BM-MSCs and that the difference is orchestrated by the AML cells. In addition this difference is regulated through PGC-1α, the master regulator of mitochondrial biogenesis. We hypothesise that therapeutic intervention targeting mitochondrial activity in the AML micro-environment would make AML more susceptible to current treatment regimes.
Session topic: E-poster
Keyword(s): Acute myeloid leukemia, Mitochondria
Type: Eposter Presentation
Background
Acute Myeloid Leukaemia (AML) is biologically a heterogeneous disease which results from defects in haematopoiesis, characterised by the accumulation of haematopoietic myeloid cells in the bone marrow. Although there have been advances in the treatment of the disease, AML remains incurable for at least 80% of patients. It has been shown that AML cells have an increased mitochondrial mass and higher levels of oxidative phosphorylation (OXPHOS) compared to normal CD34+ cells, suggesting a dependency of OXPHOS for AML survival. Little is known about the mitochondrial activity of bone marrow mesenchymal stromal cells (BM-MSC), which provide a supportive environment for AML cells in the bone marrow.
Aims
Therefore the aim of this study is to determine if there is an increase in mitochondrial respiration in BM-MSCs from AML patients (AML BM-MSCs) compared to non-AML patients (normal BM-MSCs) and to understand how the difference is regulated.
Methods
Primary AML blasts were obtained from patient bone marrow. Primary AML and normal BM-MSCs were isolated from patients bone marrow, with informed consent and under approval from the UK National Research Ethics Service (LRCEref07/H0310/146), using adherence. BM-MSC were characterised using flow cytometry for expression of CD90+, CD73+, CD105+ and CD45-. Mitochondrial respiration and glycolysis rates were measured using the Seahorse XF24 extracellular flux analyser. RT-qPCR and western blotting were used to determine peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) expression and regulation. Lentiviral mediated knockdown was also used.
Results
Our results show that AML BM-MSCs have an increased ATP production after 3 days compared to normal BM-MSCs. AML BM-MSCs also show an increased oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) compared to normal BM-MSCs. This demonstrates not only an increase in mitochondrial respiration but also increased glycolysis, which correlates with the increased ATP production of the AML BM-MSCs compared to normal BM-MSCs. Mitochondrial respiration and glycolysis increase in AML BM-MSCs after co-culture with primary AML cells. This suggests that exposure to AML cells cause the increased mitochondrial respiration and glycolysis observed in the AML BM-MSCs. There is differential expression of PGC-1α mRNA and protein in AML BM-MSCs after co-culture with AML cells, suggesting regulation of mitochondrial respiration through PGC-1α.
Conclusion
These results provide evidence of an increased mitochondrial respiration and glycolysis in the AML BM-MSCs compared to the normal BM-MSCs and that the difference is orchestrated by the AML cells. In addition this difference is regulated through PGC-1α, the master regulator of mitochondrial biogenesis. We hypothesise that therapeutic intervention targeting mitochondrial activity in the AML micro-environment would make AML more susceptible to current treatment regimes.
Session topic: E-poster
Keyword(s): Acute myeloid leukemia, Mitochondria
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