EXPLOITING OXIDATIVE PHOSPHORYLATION TO IMPROVE ANTIBODY THERAPY OF LYMPHOMA
(Abstract release date: 05/19/16)
EHA Library. Vilventhraraja E. 06/09/16; 132938; E1389
Ms. Emma Vilventhraraja
Contributions
Contributions
Abstract
Abstract: E1389
Type: Eposter Presentation
Background
Diffuse Large B Cell Lymphoma (DLBCL) is the most prevalent non-Hodgkin’s lymphoma (NHL) in adults. Since the addition of the Type I anti-CD20 antibody Rituximab to chemotherapy, overall survival of patients has improved dramatically compared to the pre-Rituximab era. However DLBCL has the worst survival rates out of all NHLs with an average 5-year survival of 55%. Unfortunately 40% of all DLBCL patients relapse within 2 years, and those that relapse or have refractory disease tend not to respond well to antibody-based salvage therapies. This has prompted many to try and enhance the efficacy of anti-CD20 antibodies in order to improve first-line treatment of DLBCL, resulting in the evolution of Type II humanised anti-CD20 antibodies. Metabolism has been reported to be involved in Type II anti-CD20 antibody-mediated cell death, however the mechanism and extent of metabolic involvement remains unknown.
Aims
The aim of this investigation is to see whether oxidative phosphorylation can be exploited to improve the efficacy of Type II anti-CD20 therapy of DLBCL.
Methods
Here, we established a panel of DLBCL cells that consisted of two groups of cell lines; one group harbouring a gene expression profile (GEP) with an enrichment of genes associated with an oxidative phosphorylation (OxPhos) phenotype, and the other group without this gene signature. We used the XF Seahorse Mito Stress Test to reveal bioenergetic profiles of the cell lines before and after treatment with a panel of anti-CD20 antibodies that included Type I and Type II anti-CD20 antibodies. This assay utilises the differential activity of four mitochondrial inhibitors to calculate basal respiration, ATP production, and maximal and spare respiratory capacity of the cell. Using the same method we then assessed whether chemically manipulating oxidative phosphorylation added to the effect on the bioenergetic profile observed following treatment with anti-CD20 antibodies. Finally, we performed clonogenic survival assays to assess whether cytotoxicity of anti-CD20 antibodies was enhanced by simultaneous treatment with Metformin, a well-established inhibitor of oxidative phosphorylation.
Results
We have observed that treatment with anti-CD20 antibodies has a significant effect on the bioenergetic profile of all DLBCL cells in our panel. Each of the antibodies in our panel had a differential ability to increase or decrease bioenergetic activity, in a cell-line specific manner. Further, we have shown that treatment with Metformin causes a significant reduction in the amount of energy produced by oxidative phosphorylation in both groups of cells in our panel. Finally, when analysing the clonogenic survival of cell lines we have found that the cytotoxicity of Type II anti-CD20 antibodies, was enhanced by simultaneously treating cell lines with an oxidative phosphorylation inhibitors.
Conclusion
With regard to clonogenicity of DLBCL cells, our data suggests that compounds that inhibit oxidative phosphorylation enhance the cytotoxicity of Type II CD20 antibodies. We believe that understanding the mechanism of loss of clonogenic survival will allow us to establish effective treatment combinations to significantly improve the efficacy of anti-CD20 antibody therapy in DLBCL.
Session topic: E-poster
Keyword(s): CD20, Cytotoxicity, DLBCL, Monoclonal antibody
Type: Eposter Presentation
Background
Diffuse Large B Cell Lymphoma (DLBCL) is the most prevalent non-Hodgkin’s lymphoma (NHL) in adults. Since the addition of the Type I anti-CD20 antibody Rituximab to chemotherapy, overall survival of patients has improved dramatically compared to the pre-Rituximab era. However DLBCL has the worst survival rates out of all NHLs with an average 5-year survival of 55%. Unfortunately 40% of all DLBCL patients relapse within 2 years, and those that relapse or have refractory disease tend not to respond well to antibody-based salvage therapies. This has prompted many to try and enhance the efficacy of anti-CD20 antibodies in order to improve first-line treatment of DLBCL, resulting in the evolution of Type II humanised anti-CD20 antibodies. Metabolism has been reported to be involved in Type II anti-CD20 antibody-mediated cell death, however the mechanism and extent of metabolic involvement remains unknown.
Aims
The aim of this investigation is to see whether oxidative phosphorylation can be exploited to improve the efficacy of Type II anti-CD20 therapy of DLBCL.
Methods
Here, we established a panel of DLBCL cells that consisted of two groups of cell lines; one group harbouring a gene expression profile (GEP) with an enrichment of genes associated with an oxidative phosphorylation (OxPhos) phenotype, and the other group without this gene signature. We used the XF Seahorse Mito Stress Test to reveal bioenergetic profiles of the cell lines before and after treatment with a panel of anti-CD20 antibodies that included Type I and Type II anti-CD20 antibodies. This assay utilises the differential activity of four mitochondrial inhibitors to calculate basal respiration, ATP production, and maximal and spare respiratory capacity of the cell. Using the same method we then assessed whether chemically manipulating oxidative phosphorylation added to the effect on the bioenergetic profile observed following treatment with anti-CD20 antibodies. Finally, we performed clonogenic survival assays to assess whether cytotoxicity of anti-CD20 antibodies was enhanced by simultaneous treatment with Metformin, a well-established inhibitor of oxidative phosphorylation.
Results
We have observed that treatment with anti-CD20 antibodies has a significant effect on the bioenergetic profile of all DLBCL cells in our panel. Each of the antibodies in our panel had a differential ability to increase or decrease bioenergetic activity, in a cell-line specific manner. Further, we have shown that treatment with Metformin causes a significant reduction in the amount of energy produced by oxidative phosphorylation in both groups of cells in our panel. Finally, when analysing the clonogenic survival of cell lines we have found that the cytotoxicity of Type II anti-CD20 antibodies, was enhanced by simultaneously treating cell lines with an oxidative phosphorylation inhibitors.
Conclusion
With regard to clonogenicity of DLBCL cells, our data suggests that compounds that inhibit oxidative phosphorylation enhance the cytotoxicity of Type II CD20 antibodies. We believe that understanding the mechanism of loss of clonogenic survival will allow us to establish effective treatment combinations to significantly improve the efficacy of anti-CD20 antibody therapy in DLBCL.
Session topic: E-poster
Keyword(s): CD20, Cytotoxicity, DLBCL, Monoclonal antibody
Abstract: E1389
Type: Eposter Presentation
Background
Diffuse Large B Cell Lymphoma (DLBCL) is the most prevalent non-Hodgkin’s lymphoma (NHL) in adults. Since the addition of the Type I anti-CD20 antibody Rituximab to chemotherapy, overall survival of patients has improved dramatically compared to the pre-Rituximab era. However DLBCL has the worst survival rates out of all NHLs with an average 5-year survival of 55%. Unfortunately 40% of all DLBCL patients relapse within 2 years, and those that relapse or have refractory disease tend not to respond well to antibody-based salvage therapies. This has prompted many to try and enhance the efficacy of anti-CD20 antibodies in order to improve first-line treatment of DLBCL, resulting in the evolution of Type II humanised anti-CD20 antibodies. Metabolism has been reported to be involved in Type II anti-CD20 antibody-mediated cell death, however the mechanism and extent of metabolic involvement remains unknown.
Aims
The aim of this investigation is to see whether oxidative phosphorylation can be exploited to improve the efficacy of Type II anti-CD20 therapy of DLBCL.
Methods
Here, we established a panel of DLBCL cells that consisted of two groups of cell lines; one group harbouring a gene expression profile (GEP) with an enrichment of genes associated with an oxidative phosphorylation (OxPhos) phenotype, and the other group without this gene signature. We used the XF Seahorse Mito Stress Test to reveal bioenergetic profiles of the cell lines before and after treatment with a panel of anti-CD20 antibodies that included Type I and Type II anti-CD20 antibodies. This assay utilises the differential activity of four mitochondrial inhibitors to calculate basal respiration, ATP production, and maximal and spare respiratory capacity of the cell. Using the same method we then assessed whether chemically manipulating oxidative phosphorylation added to the effect on the bioenergetic profile observed following treatment with anti-CD20 antibodies. Finally, we performed clonogenic survival assays to assess whether cytotoxicity of anti-CD20 antibodies was enhanced by simultaneous treatment with Metformin, a well-established inhibitor of oxidative phosphorylation.
Results
We have observed that treatment with anti-CD20 antibodies has a significant effect on the bioenergetic profile of all DLBCL cells in our panel. Each of the antibodies in our panel had a differential ability to increase or decrease bioenergetic activity, in a cell-line specific manner. Further, we have shown that treatment with Metformin causes a significant reduction in the amount of energy produced by oxidative phosphorylation in both groups of cells in our panel. Finally, when analysing the clonogenic survival of cell lines we have found that the cytotoxicity of Type II anti-CD20 antibodies, was enhanced by simultaneously treating cell lines with an oxidative phosphorylation inhibitors.
Conclusion
With regard to clonogenicity of DLBCL cells, our data suggests that compounds that inhibit oxidative phosphorylation enhance the cytotoxicity of Type II CD20 antibodies. We believe that understanding the mechanism of loss of clonogenic survival will allow us to establish effective treatment combinations to significantly improve the efficacy of anti-CD20 antibody therapy in DLBCL.
Session topic: E-poster
Keyword(s): CD20, Cytotoxicity, DLBCL, Monoclonal antibody
Type: Eposter Presentation
Background
Diffuse Large B Cell Lymphoma (DLBCL) is the most prevalent non-Hodgkin’s lymphoma (NHL) in adults. Since the addition of the Type I anti-CD20 antibody Rituximab to chemotherapy, overall survival of patients has improved dramatically compared to the pre-Rituximab era. However DLBCL has the worst survival rates out of all NHLs with an average 5-year survival of 55%. Unfortunately 40% of all DLBCL patients relapse within 2 years, and those that relapse or have refractory disease tend not to respond well to antibody-based salvage therapies. This has prompted many to try and enhance the efficacy of anti-CD20 antibodies in order to improve first-line treatment of DLBCL, resulting in the evolution of Type II humanised anti-CD20 antibodies. Metabolism has been reported to be involved in Type II anti-CD20 antibody-mediated cell death, however the mechanism and extent of metabolic involvement remains unknown.
Aims
The aim of this investigation is to see whether oxidative phosphorylation can be exploited to improve the efficacy of Type II anti-CD20 therapy of DLBCL.
Methods
Here, we established a panel of DLBCL cells that consisted of two groups of cell lines; one group harbouring a gene expression profile (GEP) with an enrichment of genes associated with an oxidative phosphorylation (OxPhos) phenotype, and the other group without this gene signature. We used the XF Seahorse Mito Stress Test to reveal bioenergetic profiles of the cell lines before and after treatment with a panel of anti-CD20 antibodies that included Type I and Type II anti-CD20 antibodies. This assay utilises the differential activity of four mitochondrial inhibitors to calculate basal respiration, ATP production, and maximal and spare respiratory capacity of the cell. Using the same method we then assessed whether chemically manipulating oxidative phosphorylation added to the effect on the bioenergetic profile observed following treatment with anti-CD20 antibodies. Finally, we performed clonogenic survival assays to assess whether cytotoxicity of anti-CD20 antibodies was enhanced by simultaneous treatment with Metformin, a well-established inhibitor of oxidative phosphorylation.
Results
We have observed that treatment with anti-CD20 antibodies has a significant effect on the bioenergetic profile of all DLBCL cells in our panel. Each of the antibodies in our panel had a differential ability to increase or decrease bioenergetic activity, in a cell-line specific manner. Further, we have shown that treatment with Metformin causes a significant reduction in the amount of energy produced by oxidative phosphorylation in both groups of cells in our panel. Finally, when analysing the clonogenic survival of cell lines we have found that the cytotoxicity of Type II anti-CD20 antibodies, was enhanced by simultaneously treating cell lines with an oxidative phosphorylation inhibitors.
Conclusion
With regard to clonogenicity of DLBCL cells, our data suggests that compounds that inhibit oxidative phosphorylation enhance the cytotoxicity of Type II CD20 antibodies. We believe that understanding the mechanism of loss of clonogenic survival will allow us to establish effective treatment combinations to significantly improve the efficacy of anti-CD20 antibody therapy in DLBCL.
Session topic: E-poster
Keyword(s): CD20, Cytotoxicity, DLBCL, Monoclonal antibody
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