BONE MARROW ADIPOCYTES PROMOTE MYELOMA CELL PROLIFERATION THROUGH INTERLEUKIN-6 SIGNALLING
(Abstract release date: 05/19/16)
EHA Library. Katumba A. 06/09/16; 132799; E1250
Mr. Alvin Katumba
Contributions
Contributions
Abstract
Abstract: E1250
Type: Eposter Presentation
Background
Multiple myeloma (MM) is a malignancy characterised by the proliferation of monoclonal plasma cells within the bone marrow (BM) and its progression crucially involves support from the BM microenvironment. The BM microenvironment consists of many cell types not directly involved in haematopoiesis including the adipocyte, which accounts for over 50% of the older adults BM by volume. Adipocytes are known to promote the growth and survival of breast, ovarian and prostate cancers. Subsequently, we hypothesised that BM adipocytes could play a role in mediating MM cell survival and proliferation.
Aims
To investigate the role of BM adipocytes in regulating the survival and proliferation of MM cells.
Methods
Primary myeloma cells and MM cell lines were cultured on patient derived adipocytes which were obtained following informed consent under approval from the UK National Research Ethics Service (LRCEref07/H0310/146). MM cell proliferation was determined using flow cytometry cell count and BrdU uptake while cell survival was determined by annexin V/PI staining. Immunocytochemistry using a lipid specific dye and nuclear staining was performed to determine free fatty acid (FFA) transfer between adipocytes and MM cells with and without the inhibitor acipimox. ß-oxidation rates in MM cells were determined by Seahorse XF analysis using palmitate and etomoxir. PCR and cytokine analysis was used to determine the levels of interleukin-6 (IL-6) and other factors.
Results
We report that BM adipocytes promote MM cell survival and proliferation. BrdU analysis and annexin V/PI staining revealed that adipocyte/MM cell co-culture led to increased MM cell proliferation and decreased MM cell apoptosis compared to MM cells cultured alone. Furthermore, MM cells cultured on adipocytes induced adipocyte lipolysis and initiated lipid transfer from adipocytes to MM cells. This process was inhibited by acipimox. In addition, MM cell/adipocyte co-culture also led to increased MM cell β-oxidation rates indicating that MM cells use BM adipocyte derived FFAs to produce ATP. Furthermore, pharmacological inhibition of ß-oxidation using etomoxir led to a significant decrease in survival and proliferation of MM cells. PCR and cytokine analysis identified an increased expression of IL-6 in MM cells co-cultured with adipocytes. Further analysis revealed that IL-6 derived from myeloma cells induced the breakdown of triglycerides into FFAs (lipolysis) in BM adipocytes.
Conclusion
Results indicate that adipocytes provide energy to MM cells in the form of FFAs for survival and growth in vitro. Moreover, we found that the signal which induces adipocyte lipolysis is myeloma derived IL-6.
Session topic: E-poster
Keyword(s): Myeloma, Progression, Proliferation
Type: Eposter Presentation
Background
Multiple myeloma (MM) is a malignancy characterised by the proliferation of monoclonal plasma cells within the bone marrow (BM) and its progression crucially involves support from the BM microenvironment. The BM microenvironment consists of many cell types not directly involved in haematopoiesis including the adipocyte, which accounts for over 50% of the older adults BM by volume. Adipocytes are known to promote the growth and survival of breast, ovarian and prostate cancers. Subsequently, we hypothesised that BM adipocytes could play a role in mediating MM cell survival and proliferation.
Aims
To investigate the role of BM adipocytes in regulating the survival and proliferation of MM cells.
Methods
Primary myeloma cells and MM cell lines were cultured on patient derived adipocytes which were obtained following informed consent under approval from the UK National Research Ethics Service (LRCEref07/H0310/146). MM cell proliferation was determined using flow cytometry cell count and BrdU uptake while cell survival was determined by annexin V/PI staining. Immunocytochemistry using a lipid specific dye and nuclear staining was performed to determine free fatty acid (FFA) transfer between adipocytes and MM cells with and without the inhibitor acipimox. ß-oxidation rates in MM cells were determined by Seahorse XF analysis using palmitate and etomoxir. PCR and cytokine analysis was used to determine the levels of interleukin-6 (IL-6) and other factors.
Results
We report that BM adipocytes promote MM cell survival and proliferation. BrdU analysis and annexin V/PI staining revealed that adipocyte/MM cell co-culture led to increased MM cell proliferation and decreased MM cell apoptosis compared to MM cells cultured alone. Furthermore, MM cells cultured on adipocytes induced adipocyte lipolysis and initiated lipid transfer from adipocytes to MM cells. This process was inhibited by acipimox. In addition, MM cell/adipocyte co-culture also led to increased MM cell β-oxidation rates indicating that MM cells use BM adipocyte derived FFAs to produce ATP. Furthermore, pharmacological inhibition of ß-oxidation using etomoxir led to a significant decrease in survival and proliferation of MM cells. PCR and cytokine analysis identified an increased expression of IL-6 in MM cells co-cultured with adipocytes. Further analysis revealed that IL-6 derived from myeloma cells induced the breakdown of triglycerides into FFAs (lipolysis) in BM adipocytes.
Conclusion
Results indicate that adipocytes provide energy to MM cells in the form of FFAs for survival and growth in vitro. Moreover, we found that the signal which induces adipocyte lipolysis is myeloma derived IL-6.
Session topic: E-poster
Keyword(s): Myeloma, Progression, Proliferation
Abstract: E1250
Type: Eposter Presentation
Background
Multiple myeloma (MM) is a malignancy characterised by the proliferation of monoclonal plasma cells within the bone marrow (BM) and its progression crucially involves support from the BM microenvironment. The BM microenvironment consists of many cell types not directly involved in haematopoiesis including the adipocyte, which accounts for over 50% of the older adults BM by volume. Adipocytes are known to promote the growth and survival of breast, ovarian and prostate cancers. Subsequently, we hypothesised that BM adipocytes could play a role in mediating MM cell survival and proliferation.
Aims
To investigate the role of BM adipocytes in regulating the survival and proliferation of MM cells.
Methods
Primary myeloma cells and MM cell lines were cultured on patient derived adipocytes which were obtained following informed consent under approval from the UK National Research Ethics Service (LRCEref07/H0310/146). MM cell proliferation was determined using flow cytometry cell count and BrdU uptake while cell survival was determined by annexin V/PI staining. Immunocytochemistry using a lipid specific dye and nuclear staining was performed to determine free fatty acid (FFA) transfer between adipocytes and MM cells with and without the inhibitor acipimox. ß-oxidation rates in MM cells were determined by Seahorse XF analysis using palmitate and etomoxir. PCR and cytokine analysis was used to determine the levels of interleukin-6 (IL-6) and other factors.
Results
We report that BM adipocytes promote MM cell survival and proliferation. BrdU analysis and annexin V/PI staining revealed that adipocyte/MM cell co-culture led to increased MM cell proliferation and decreased MM cell apoptosis compared to MM cells cultured alone. Furthermore, MM cells cultured on adipocytes induced adipocyte lipolysis and initiated lipid transfer from adipocytes to MM cells. This process was inhibited by acipimox. In addition, MM cell/adipocyte co-culture also led to increased MM cell β-oxidation rates indicating that MM cells use BM adipocyte derived FFAs to produce ATP. Furthermore, pharmacological inhibition of ß-oxidation using etomoxir led to a significant decrease in survival and proliferation of MM cells. PCR and cytokine analysis identified an increased expression of IL-6 in MM cells co-cultured with adipocytes. Further analysis revealed that IL-6 derived from myeloma cells induced the breakdown of triglycerides into FFAs (lipolysis) in BM adipocytes.
Conclusion
Results indicate that adipocytes provide energy to MM cells in the form of FFAs for survival and growth in vitro. Moreover, we found that the signal which induces adipocyte lipolysis is myeloma derived IL-6.
Session topic: E-poster
Keyword(s): Myeloma, Progression, Proliferation
Type: Eposter Presentation
Background
Multiple myeloma (MM) is a malignancy characterised by the proliferation of monoclonal plasma cells within the bone marrow (BM) and its progression crucially involves support from the BM microenvironment. The BM microenvironment consists of many cell types not directly involved in haematopoiesis including the adipocyte, which accounts for over 50% of the older adults BM by volume. Adipocytes are known to promote the growth and survival of breast, ovarian and prostate cancers. Subsequently, we hypothesised that BM adipocytes could play a role in mediating MM cell survival and proliferation.
Aims
To investigate the role of BM adipocytes in regulating the survival and proliferation of MM cells.
Methods
Primary myeloma cells and MM cell lines were cultured on patient derived adipocytes which were obtained following informed consent under approval from the UK National Research Ethics Service (LRCEref07/H0310/146). MM cell proliferation was determined using flow cytometry cell count and BrdU uptake while cell survival was determined by annexin V/PI staining. Immunocytochemistry using a lipid specific dye and nuclear staining was performed to determine free fatty acid (FFA) transfer between adipocytes and MM cells with and without the inhibitor acipimox. ß-oxidation rates in MM cells were determined by Seahorse XF analysis using palmitate and etomoxir. PCR and cytokine analysis was used to determine the levels of interleukin-6 (IL-6) and other factors.
Results
We report that BM adipocytes promote MM cell survival and proliferation. BrdU analysis and annexin V/PI staining revealed that adipocyte/MM cell co-culture led to increased MM cell proliferation and decreased MM cell apoptosis compared to MM cells cultured alone. Furthermore, MM cells cultured on adipocytes induced adipocyte lipolysis and initiated lipid transfer from adipocytes to MM cells. This process was inhibited by acipimox. In addition, MM cell/adipocyte co-culture also led to increased MM cell β-oxidation rates indicating that MM cells use BM adipocyte derived FFAs to produce ATP. Furthermore, pharmacological inhibition of ß-oxidation using etomoxir led to a significant decrease in survival and proliferation of MM cells. PCR and cytokine analysis identified an increased expression of IL-6 in MM cells co-cultured with adipocytes. Further analysis revealed that IL-6 derived from myeloma cells induced the breakdown of triglycerides into FFAs (lipolysis) in BM adipocytes.
Conclusion
Results indicate that adipocytes provide energy to MM cells in the form of FFAs for survival and growth in vitro. Moreover, we found that the signal which induces adipocyte lipolysis is myeloma derived IL-6.
Session topic: E-poster
Keyword(s): Myeloma, Progression, Proliferation
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