Contributions
Abstract: PB2327
Type: Publication Only
Background
Lymphomas are a heterogeneous group of lymphoproliferative disorders of B and T cell origin that are treated with chemotherapy drugs with variable success rate that has virtually not changed over decades due to their more complex nature than solid malignancies. Till date new classes of chemotherapy and metabolic drugs have emerged, however durable responses to these conventional and new therapies are achieved in a limited number of cancer patients, with many individuals experiencing resistance to the drugs.
Aims
The paucity in our understanding of what regulates the drug resistance phenotype and establishing a predictive indicator is, in great part, due to the lack of adequate ex vivo lymphoma organoids models to accurately study the effect of microenvironmental signaling in which Non Hodgkin lymphomas cells arise and properly studied. Therefore, is a need to develop 3D tissues models that mimic the NHL microenvironment model for NHL biology and drug responses.
Methods
3D cells culture methods including hydrogel, viability assay like WST based analysis, Annexin/PI staining, RT-PCRs for genes analysis, confocal microscopy and western blot analysis for protein expression analysis, Flow cytometric analysis for single cells sorting and cancer stem cell analysis, knock down effects by using specific inhibitions or designing siRNA of specific signaling molecule.
Results
we explored strategies to enhance chemosensitivity to doxorubicin, an important chemotherapeutic drug widely used for the treatment of hematological malignancies. Lymphoma cells grown in this model exhibited excellent biomimetic properties compared to conventional 2D culture including (1) enhanced chemotherapy resistance, (2) suppressed rate of apoptosis, (3) upregulated expression of drug resistance genes (MDR1, MRP1, BCRP and HIF- 1α), (4) elevated levels of tumor aggressiveness factors including Notch (Notch-1, -2, -3, and -4) and its downstream molecules (Hes-1 and Hey-1), VEGF and MMPs (MMP-2 and MMP-9), and (5) enrichment of a lymphoma stem cell population. Tiam1, a potential biomarker of tumor progression, metastasis, and chemoresistance, was activated in our 3D lymphoma model. Remarkably, we identified two synergistic therapeutic oncotargets, Tiam1 and Notch, as a strategy to combat resistance against doxorubicin in EL4 T and A20 B lymphoma.
Conclusion
Therefore, our data suggest that our 3D lymphoma model is a promising in vitro research platform for studying lymphoma biology and therapeutic approaches.
Session topic: 19. Non-Hodgkin lymphoma Biology & Translational Research
Keyword(s): Doxorubicin
Abstract: PB2327
Type: Publication Only
Background
Lymphomas are a heterogeneous group of lymphoproliferative disorders of B and T cell origin that are treated with chemotherapy drugs with variable success rate that has virtually not changed over decades due to their more complex nature than solid malignancies. Till date new classes of chemotherapy and metabolic drugs have emerged, however durable responses to these conventional and new therapies are achieved in a limited number of cancer patients, with many individuals experiencing resistance to the drugs.
Aims
The paucity in our understanding of what regulates the drug resistance phenotype and establishing a predictive indicator is, in great part, due to the lack of adequate ex vivo lymphoma organoids models to accurately study the effect of microenvironmental signaling in which Non Hodgkin lymphomas cells arise and properly studied. Therefore, is a need to develop 3D tissues models that mimic the NHL microenvironment model for NHL biology and drug responses.
Methods
3D cells culture methods including hydrogel, viability assay like WST based analysis, Annexin/PI staining, RT-PCRs for genes analysis, confocal microscopy and western blot analysis for protein expression analysis, Flow cytometric analysis for single cells sorting and cancer stem cell analysis, knock down effects by using specific inhibitions or designing siRNA of specific signaling molecule.
Results
we explored strategies to enhance chemosensitivity to doxorubicin, an important chemotherapeutic drug widely used for the treatment of hematological malignancies. Lymphoma cells grown in this model exhibited excellent biomimetic properties compared to conventional 2D culture including (1) enhanced chemotherapy resistance, (2) suppressed rate of apoptosis, (3) upregulated expression of drug resistance genes (MDR1, MRP1, BCRP and HIF- 1α), (4) elevated levels of tumor aggressiveness factors including Notch (Notch-1, -2, -3, and -4) and its downstream molecules (Hes-1 and Hey-1), VEGF and MMPs (MMP-2 and MMP-9), and (5) enrichment of a lymphoma stem cell population. Tiam1, a potential biomarker of tumor progression, metastasis, and chemoresistance, was activated in our 3D lymphoma model. Remarkably, we identified two synergistic therapeutic oncotargets, Tiam1 and Notch, as a strategy to combat resistance against doxorubicin in EL4 T and A20 B lymphoma.
Conclusion
Therefore, our data suggest that our 3D lymphoma model is a promising in vitro research platform for studying lymphoma biology and therapeutic approaches.
Session topic: 19. Non-Hodgkin lymphoma Biology & Translational Research
Keyword(s): Doxorubicin