- Email: [email protected]
New Targeted Drugs in Multiple Myeloma Therapy - in Vitro Studies
Abstracts the patients (24%) were obese (body mass index >30 Kg/M2) and 43/82 (52%) were overweight (body mass index > 26 Kg/M2). 8/ 82 patients 9.7%) had a raised HbA1c (> 48mmol/mol) consistent with diabetes mellitus and a further 4 patients (5%) had HbA1c suggestive of insulin resistance (>42 mmol/mol). In this patient cohort 58.5% of patients had at least 1 feature consistent with metabolic syndrome (as defined by: diabetes or insulin resistance, triglycerides >1.7mmol/l, HDL-cholesterol<1mmol/l, BMI>30Kg/m2 or BP>140/90). Conclusion: We conclude that a notable proportion of our patients with myeloma had features of metabolic syndrome including obesity. Whether these features of metabolic syndrome are causative of myeloma or are associated with survival merits further study.
PO-308 New Targeted Drugs in Multiple Myeloma Therapy - in Vitro Studies C. Geraldes, A.C. Gonçalves, R. Alves, A. Roque, J. Manuel, N. Costa, A.B. Sarmento-Ribeiro Clinical Haematology Department, Coimbra University Hospital Center, Coimbra, Portugal; University Clinic of Hematology, Coimbra,
the IC50 was 17,5 M for EVE, 150 M for SLB and 12,5 M for PTN. We also observed that each one of these drugs increased the cytotoxic effect of BTZ when combined with low doses of this drug. These compounds induced cell death through apoptosis, which may be mediated by the observed increase in the expression of activated caspases. Besides the cytotoxic effect, EVE showed also an antiproliferative effect once this drug induced a G0/G1 arrest in MM cells. Conclusions: Our results suggest that novel drugs with innovative mechanisms of action might potentiate the activity of BTZ and increase the efficacy of MM treatment.
PO-309 The relationship of the parameters of myeloma bone disease signalling pathways to the extent of bone involvement and extramedullary disease in monoclonal gammopathies J. Minarik,1 Z. Hermanova,2 P. Petrova,3 J. Zapletalova,4 J. Hrbek,5 P. Pusciznova,1 J. Bacovsky,1 T. Pika,1 M. Herman,5 V. Scudla1
Portugal; Centre for Research on Environment, Genetics and
1
Oncobiology, Coimbra, Portugal; Applied Molecular Biology and University Clinic of Hematology, Coimbra, Portugal; Faculty of Medi-
Palacky University Olomouc and University Hospital Olomouc; 2 Department of Immunology, Faculty of Medicine and Dentistry, Pal-
cine, University of Coimbra, Coimbra, Portugal
acky University Olomouc and University Hospital Olomouc; 3Depart-
Department of Hemato-Oncology, Faculty of Medicine and Dentistry,
ment of Clinical Biochemistry, Faculty of Medicine and Dentistry,
Background: Multiple Myeloma (MM) remains an incurable disease, despite all recent therapeutic advances and new challenging strategies are needed. Bortezomib (BTZ), a first generation proteasome inhibitor, is a milestone in MM treatment. Parthenolide (PTN), a sesquiterpene lactone, has been demonstrated to induce apoptosis in cancer cells from several tumors. PTN inhibits NF B signaling pathway, constitutively activated in MM cells, and whose activity is also inhibited by BTZ. Silibinin (SLB) induces both caspase-dependent and -independent apoptosis and downregulates survivin. Everolimus (EVE), a mammalian target of rapamycin (mTOR) inhibitor, blocks protein translation related to cell survival and cell proliferation. These three drugs have already shown an anticancer activity in various tumor cell types; however in MM studies are scarce. Aims: Our aim was to investigate the cytotoxic and antiproliferative effects of PTN, SLB and EVE either alone or in combination with BTZ, in a MM cell line, in order to identify new therapeutic options in MM. Methods: For this purpose, NCI H929 cells (an in vitro model of MM) were incubated in the absence and in the presence of increasing concentrations of PTN, SLB and EVE, in monotherapy and in combination with BTZ, at different time points, and in several combination schedules. Cell viability was analysed by Resazurin assays. Cell death was determined by optical microscopy, after staining the smears with MayGrünwald-Giemsa, and by flow cytometry using annexin-V and propidium-iodide double staining. By flow cytometry we also evaluated caspases activation, using the apostat probe, and cell cycle analysis, using propidium-iodide. Results: Our results showed that EVE, PTN and SLB induce a decrease in cell viability in a dose- and time-dependent manner. Indeed, after 72 hours of cell treatment,
e254
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15th International Myeloma Workshop, September 23-26, 2015
Palacky University Olomouc and University Hospital Olomouc; 4
Department of Medical Biophysics, Faculty of Medicine and
Dentistry, Palacky University Olomouc and University Hospital Olomouc; 5Department of Radiology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc
Introduction: We carried out prospective assessment of the relationship of selected parameters of signalling pathways in myeloma bone disease (MBD) to the extent of skeletal involvement and to the presence of extramedullary disease in patients with monoclonal gammopathies. Patients and Methods: We assessed 77 patients with monoclonal gammopathies: 46 patients with active multiple myeloma, 12 with smoldering myeloma and 19 individuals with monoclonal gammopathy of undetermined significance. In the patients we assessed selected parameters of MBD signalling with regard to the extent of myeloma bone disease evaluated using whole body magnetic resonance and low-dose computed tomography. In addition, we addressed potential relationship to the presence of extramedullary involvement. We assessed following parameters: HGF, macrophage inflammatory factor alpha (MIP-1 ), syndecan-1, osteoprotegerin (OPG), Activin A, DKK1, Annexin A2 and NF- B. For statistical estimation we used Kruskal-Wallis test and Mann-Whitney U post hoc test with Bonferroni correction at p < 0,05. Results: Due to uneven distribution of bone involvement we assessed the extreme situations e individuals without skeletal involvement and patients with extensive bone involvement. Between these two groups we found statistically significant difference inserum levels of MIP-1 (median ¼ M 21,8 vs 25,8pg/ml, p ¼ 0,003), syndecan-1 (M 29,7
PO-308 New Targeted Drugs in Multiple Myeloma Therapy - in Vitro Studies C. Geraldes, A.C. Gonçalves, R. Alves, A. Roque, J. Manuel, N. Costa, A.B. Sarmento-Ribeiro Clinical Haematology Department, Coimbra University Hospital Center, Coimbra, Portugal; University Clinic of Hematology, Coimbra,
the IC50 was 17,5 M for EVE, 150 M for SLB and 12,5 M for PTN. We also observed that each one of these drugs increased the cytotoxic effect of BTZ when combined with low doses of this drug. These compounds induced cell death through apoptosis, which may be mediated by the observed increase in the expression of activated caspases. Besides the cytotoxic effect, EVE showed also an antiproliferative effect once this drug induced a G0/G1 arrest in MM cells. Conclusions: Our results suggest that novel drugs with innovative mechanisms of action might potentiate the activity of BTZ and increase the efficacy of MM treatment.
PO-309 The relationship of the parameters of myeloma bone disease signalling pathways to the extent of bone involvement and extramedullary disease in monoclonal gammopathies J. Minarik,1 Z. Hermanova,2 P. Petrova,3 J. Zapletalova,4 J. Hrbek,5 P. Pusciznova,1 J. Bacovsky,1 T. Pika,1 M. Herman,5 V. Scudla1
Portugal; Centre for Research on Environment, Genetics and
1
Oncobiology, Coimbra, Portugal; Applied Molecular Biology and University Clinic of Hematology, Coimbra, Portugal; Faculty of Medi-
Palacky University Olomouc and University Hospital Olomouc; 2 Department of Immunology, Faculty of Medicine and Dentistry, Pal-
cine, University of Coimbra, Coimbra, Portugal
acky University Olomouc and University Hospital Olomouc; 3Depart-
Department of Hemato-Oncology, Faculty of Medicine and Dentistry,
ment of Clinical Biochemistry, Faculty of Medicine and Dentistry,
Background: Multiple Myeloma (MM) remains an incurable disease, despite all recent therapeutic advances and new challenging strategies are needed. Bortezomib (BTZ), a first generation proteasome inhibitor, is a milestone in MM treatment. Parthenolide (PTN), a sesquiterpene lactone, has been demonstrated to induce apoptosis in cancer cells from several tumors. PTN inhibits NF B signaling pathway, constitutively activated in MM cells, and whose activity is also inhibited by BTZ. Silibinin (SLB) induces both caspase-dependent and -independent apoptosis and downregulates survivin. Everolimus (EVE), a mammalian target of rapamycin (mTOR) inhibitor, blocks protein translation related to cell survival and cell proliferation. These three drugs have already shown an anticancer activity in various tumor cell types; however in MM studies are scarce. Aims: Our aim was to investigate the cytotoxic and antiproliferative effects of PTN, SLB and EVE either alone or in combination with BTZ, in a MM cell line, in order to identify new therapeutic options in MM. Methods: For this purpose, NCI H929 cells (an in vitro model of MM) were incubated in the absence and in the presence of increasing concentrations of PTN, SLB and EVE, in monotherapy and in combination with BTZ, at different time points, and in several combination schedules. Cell viability was analysed by Resazurin assays. Cell death was determined by optical microscopy, after staining the smears with MayGrünwald-Giemsa, and by flow cytometry using annexin-V and propidium-iodide double staining. By flow cytometry we also evaluated caspases activation, using the apostat probe, and cell cycle analysis, using propidium-iodide. Results: Our results showed that EVE, PTN and SLB induce a decrease in cell viability in a dose- and time-dependent manner. Indeed, after 72 hours of cell treatment,
e254
-
15th International Myeloma Workshop, September 23-26, 2015
Palacky University Olomouc and University Hospital Olomouc; 4
Department of Medical Biophysics, Faculty of Medicine and
Dentistry, Palacky University Olomouc and University Hospital Olomouc; 5Department of Radiology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc
Introduction: We carried out prospective assessment of the relationship of selected parameters of signalling pathways in myeloma bone disease (MBD) to the extent of skeletal involvement and to the presence of extramedullary disease in patients with monoclonal gammopathies. Patients and Methods: We assessed 77 patients with monoclonal gammopathies: 46 patients with active multiple myeloma, 12 with smoldering myeloma and 19 individuals with monoclonal gammopathy of undetermined significance. In the patients we assessed selected parameters of MBD signalling with regard to the extent of myeloma bone disease evaluated using whole body magnetic resonance and low-dose computed tomography. In addition, we addressed potential relationship to the presence of extramedullary involvement. We assessed following parameters: HGF, macrophage inflammatory factor alpha (MIP-1 ), syndecan-1, osteoprotegerin (OPG), Activin A, DKK1, Annexin A2 and NF- B. For statistical estimation we used Kruskal-Wallis test and Mann-Whitney U post hoc test with Bonferroni correction at p < 0,05. Results: Due to uneven distribution of bone involvement we assessed the extreme situations e individuals without skeletal involvement and patients with extensive bone involvement. Between these two groups we found statistically significant difference inserum levels of MIP-1 (median ¼ M 21,8 vs 25,8pg/ml, p ¼ 0,003), syndecan-1 (M 29,7