- Email: [email protected]
Indirubins: A Potential Therapeutic Target in Multiple Myeloma
Abstracts we analyzed the characteristics of patients in whom serum sSLAMF7 was detected and the correlation with SLAMF7 expression levels in CD138+ myeloma cells. Finally, NK cell-mediated antibody-mediated antibody-dependent cytotoxic (ADCC) activity against the SLAM7-expressing MM cell line U266 was assessed in the lactate dehydrogenase (LDH)-based cytotoxicity assay using NK cell line NK-92MI. Results: 33% of MM patients (n¼34), but no MGUS patients or controls, had detectable sSLAMF7 in serum (range: 0.011e14.7 ng/mL). sSLAMF7 levels in symptomatic MM patients were significantly higher than those in asymptomatic ones (P¼0.0316). Furthermore, sSLAMF7 levels in patients with International Staging System (ISS) and revised ISS (R-ISS) stage 2/3 were markedly increased in comparison with ISS and R-ISS stage 1 (P¼0.0007 and 0.0177, respectively). Serum sSLAMF7 concentrations were not correlated with the percentage of plasma cells or SLAMF7 mRNA expression levels in CD138+ plasma cells in BM obtained from patients. When comparing the clinical characteristics between sSLAMF7-positive (n¼34) and -negative groups (n¼69), plasma cell percentage and LDH levels, and corrected calcium, creatinine, C-reactive protein, b2-microglobulin, and interleukin-6 levels were significantly higher but levels of hemoglobin and albumin and estimated glomerular filtration rate were markedly lower in the sSLAMF7-positive group. In the in vitro study, NK cellmediated ADCC activity using anti-SLAMF7 antibody was inhibited by recombinant SLAMF7. Conclusion: Our data suggest that high levels of serum sSLAMF7 in MM patients are correlated with advanced disease, and therefore sSLAMF7 levels may be a useful indicator of disease progression in MM. Moreover, sSLAMF7 may suppress the therapeutic effects of elotuzumab in MM. Further studies are in progress to clarify the mechanism of sSLAMF7 production in MM.
PS-068 (d) Growth Differentiation Factor 15 (GDF-15) Is a New Biomarker for Overall Survival and Renal Outcomes in Patients with Light Chain (AL) Amyloidosis 1
PS-069 2
Efstathios Kastritis, Ioannis Papassotiriou, Giampaolo Merlini,3 Paolo Milani,4 Evangelos Terpos5 1
University of Athens, Athens; 2Aghia Sofia Chldren’s Hospital,
Athens; 3University Hospital Policlinico San Matteo, Pavia, Italy; 4
Amyloidosis Research and Treatment Center, Fondazione IRCCS
Policlinico San Matteo and University o, Pavia, Italy; 5National and Kapodistrian University of Athens, School of Medicine, Greece
Growth differentiation factor-15 (GDF-15), a member of TGFbeta family, is involved in several pathological conditions, including inflammation, cancer, cardiovascular and renal diseases. Serum GDF-15 levels adds prognostic information to conventional prognostic factors, such as NT-proBNP and troponins, in cardiovascular disorders and has been associated with risk of end stage renal disease in diabetics. We evaluated the prognostic significance of serum GDF-15 in two independent cohorts from the Pavia Amyloidosis
e40
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16th International Myeloma Workshop March 1-4, 2017
Center and the Department of Clinical Therapeutics, Athens. Serum levels of GDF-15 were measured by a novel pre-commercial immunoassay (Roche Diagnostics) in stored serum. The Pavia cohort included 202 and the Athens cohort included 107 patients with AL amyloidosis. Median age and involved FLC levels were similar between the two cohorts but there were differences in other baseline characteristics including heart involvement (77% vs 62% for Pavia vs Athens, p¼0.007), Mayo stage 3(42% in Pavia vs 31% in Athens cohort, p¼0.04), but stage 3B was similar (22% vs 24%) and peripheral nerve involvement (11% in Pavia vs 25% in Athens cohort, p¼0.001). Renal involvement (67% vs 72%, p¼0.415), median eGFR and renal stage distribution were similar (p¼0.544). Two year survival was 59% for Pavia and 56% for Athens cohort. Median GDF-15 levels was 3027 pg/ml in Pavia and 3854 pg/ml in Athens cohort (p¼0.09). The upper quartile of GDF-15 levels, however, was 5658 pg/ml in the Pavia and 7553 pg/ml in Athens cohort, and 90% and 94% of patients in the two cohorts had GDF-15 levels >1200 pg/ml (the upper limit of normal for individuals without cardiovascular disease). We evaluated the prognostic significance of GDF-15 levels for survival in the two cohorts by applying the previously identified cutoff of 7575 pg/ml. GDF-15 above cutoff was associated with shorter survival both in Pavia (17 months vs not reached, p¼0.003) and Athens cohort (13 vs 47 months, p¼0.03). In separate multivariate models for each cohort, that included Mayo stage, GDF-15 remained of independent prognostic significance over Mayo stage, with a hazard ratio (HR) of 1.9 (95% CI 1.2-3.3, p¼0.01) in Pavia cohort and a HR of 1.8 (95% CI 1.2-3.6, p¼0.03) in the Athens cohort. We then evaluated GDF-15 levels for prognosis of renal outcomes (dialysis): GDF-15 >4000 pg/ml was associated with a HR of 7 (95% CI 2015.6, p¼0.001) in Athens cohort (progression to dialysis within 2 years in 7% vs 47%); and HR of 4.9 in Pavia cohort. In both cohorts GDF-15 levels were prognostic for dialysis independent renal stage. Thus, we validated and confirmed in two independent cohorts, with differences in their characteristics, the prognostic value of GDF-15, as a biomarker with prognostic implications for different outcomes in patients with AL amyloidosis. Importantly, GDF-15 emerges also as new biomarkers for renal outcomes in patients with AL amyloidosis.
Indirubins: A Potential Therapeutic Target in Multiple Myeloma Tina Bagratuni,1 Nicolas Gaboriad-Kolar,2 Roubini Zakopoulou,2 Vassilios Myrianthopoulos,2 Efstathios Kastritis,3 Evangelos Terpos,3 Emmanuel Mikros,2 Alexios-Leandros Skaltsounis,2 Meletios Athanasios Dimopoulos4 1 National and Kapodistrian University of Athens, School of Medicine, Athens, Greece; 2National and Kapodistrian University of Athens; 3
national and Kapodistrian University of Athens, School of Medicine,
Greece; 4National and Kapodistrian University of Athens, School of Medicine, Athens, Greece
Current drugs in the treatment of Multiple Myeloma (MM) result in cell death via a number of mechanisms including a direct effect on plasma cells as well as alteration in the BM microenvironment.
Abstracts Among the FDA approved kinase inhibitors, few are based on natural scaffolds. 6-bromoindirubin-3’-oxime (6BIO) is a potent kinase inhibitor based on the natural 6-bromoindirubin scaffold. Indirubin and 6-bromoindirubin are two natural products that have found a particular interest in dye chemistry as the main constituent of indigo and Tyrian purple dyes. Recent findings discovered that 6BIO was a promising anti-cancer agent acting on the JAK/STAT signaling pathway mediating cell proliferation. A library containing 2000 natural molecules was constructed using several data platforms. Each molecule was processed through different filters such as tautomeric studies, protonation and steroisomerism status. Two approaches were followed: the structure-based virtual screening and the ligand-based virtual screening. The results of both approaches were combined, the molecules ranked, and 100 out of 2000 were identified as strong potential bioactive hits for the b5 subunit. Out of these 100 molecules, the chemical structures of high interest were the following: indole alkaloids derivatives (indirubins), flavonoids, secoiridoids, simple phenolic acids and acetophenone. Fifty indirubins derivatives were selected based on different criteria: structure, known/unknown targets, chemodiversity in substitution patterns. To explore the inhibitory effects of indirubins in MM, we performed the WST1 proliferation assay in three MM cell lines (H929, JJN3, L363). Initially, all the selected indirubins(w50 indirubins) were tested at 7.5mM in L363 cell line and proliferation results from the WST1 assay extracted after 24 hours of treatment. More than half of the indirubins tested displayed more than 50% reduction of the proliferation at 7.5mM. Interestingly, 10 out of the 50 indirubinstested reduced more than 80% proliferation after 24 hours. The most active indirubins were tested in H929 and JJN3 cell lines, where similar effects were seen after 24 hours of treatment. All tested indirubins acted in a dose-dependent manner. Among the most active indirubins, two molecules namely 805 and 673 emerged as attractive for further development. Compound 805 is an analog of MLS-2384 while compound 673 is an analog of MLS-2438. The latter derivative represents a promising candidate displaying an IC50 below the micromolar range on H929 and JJN3 cells. To determine the kind of cell death caused by one of the most active indirubins, 673, cell cycle analysis was performed before and after treatment in H929 cell line. In particular changes in RNA expression of 84 genes key to cell cycle regulation were analyzed in H929 cell line. Our results show that among other genes, the ones which have a dramatic increase in their expression.
Primary plasma cell leukemia (pPCL) exhibits poor outcome. We and others have previously demonstrated that novel agents and mainly bortezomib-based regimens (BBR) improve response rates and survival, however the prognostic impact of current treatments and biological markers on the outcome of pPCL has not been sufficiently explored, outside clinical trials. We analyzed the medical records of 50 patients with pPCL (M/F: 25:25; median age 65.5 years, range: 32-86 years; IgG: 19, IgA: 9, light-chain: 14, IgD: 2, non-secretory: 6; ISS1: 5, ISS2: 16, ISS3: 29) out of 2711 myeloma patients (1.8%), registered in the Greek Myeloma study group database, between 2000-2015; 52% of patients had Eastern Cooperative Group (ECOG) performance status 2; 53% had abnormal lactate dehydrogenase (LDH), 28% had hypercalcemia, 68% had hemoglobin <10 g/dL and 65% had high risk cytogenetics; 49/50 patients received therapy: 38 patients received BBR, one patient received melphalan-prednisone-thalidomide and 10 patients received conventional chemotherapy (C/T); 15 patients underwent ASCT consolidation; 48 patients were evaluated for response: 79% achieved objective response (PR) and 35% had at least very good partial response (vgPR), including 17% complete responses. Achievement of vgPR correlated with BBR+ASCT (p¼0.02). After a median follow up of 61 months (95% CI: 34.587.4), 38 patients have died (disease progression: 18, infection: 16, other causes: 4). Early mortality (1 month) occurred in 3/38 deceased patients; 31/38 patients who responded to treatment progressed and 27/31 received 2nd line treatment (lenalidomidebased: 7, BBR: 16, C/T: 4). Progression-free survival was 12 months (95% CI: 8.5-15.4) and it was marginally longer in patients treated with BBR+ASCT vs. others (18 vs. 10 months, p¼0.07). Median OS was 17 months (95% CI: 13-21 months) and it was double in patients treated with BBR+ASCT compared to others (33 months vs. 16 months); median survival after PCL progression was 7 months (95% CI: 3-11 months).
PS-071 Exome Sequencing of AL Amyloidosis Reveals Recurrently Mutated Genes Zuzana Kufova,1 Tereza Sevcikova,2 Petr Vojta,3 Jana Filipova,4 Katerina Growkova,5 Sebastian Grosicki,6 Fedor Kryukov,5 Roman Hajek7 1
Univesity Hospital Ostrava, Ostrava-Poruba, Czech Republic;
PS-070
2
Real-World Data on Clinical Characteristics, Prognosis and Outcome of Primary Plasma Cell Leukemia: A Study of the Greek Myeloma Study Group in the Era of Novel Agents 1
2
Faculty of Medicine, University of Ostrava, Ostrava, Ostrava Zábr eh, Czech Republic; 3Institute of Molecular and Translational Medicine, Palacky University in Olomouc, Olomouc; 4Department of Haemato-Oncology, Univesity Hospital Ostrava, Ostrava-Poruba, Czech Republic; 5Faculty of Medicine, University of Ostrava, Ostrava, Ostrava - Zábr eh; 6Silesian Medical University, Katowice, Poland; 7
University Hospital Ostrava and Faculty of Medicine Ostrava,
Eirini Katodritou, Evangelos Terpos, Sossana Delimpasi,3 Maria Kotsopoulou4
Ostrava, Czech Republic
1
Background: Immunoglobulin light chain amyloidosis (ALA) is a plasma cell dyscrasia characterized by deposition of amyloid fibrils in various organs and tissues, leading to organ dysfunction. Until now, only one systematic study of genomic profile of ALA on exome
Theagenion Cancer Hospital, Thessaloniki, Greece; 2National and
Kapodistrian University of Athens, School of Medicine, Greece; 3
Evangelismos Hospital, Athens, Greece; 4Metaxa Cancer Hospital,
PIRAEUS, Greece
16th International Myeloma Workshop March 1-4, 2017
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PS-068 (d) Growth Differentiation Factor 15 (GDF-15) Is a New Biomarker for Overall Survival and Renal Outcomes in Patients with Light Chain (AL) Amyloidosis 1
PS-069 2
Efstathios Kastritis, Ioannis Papassotiriou, Giampaolo Merlini,3 Paolo Milani,4 Evangelos Terpos5 1
University of Athens, Athens; 2Aghia Sofia Chldren’s Hospital,
Athens; 3University Hospital Policlinico San Matteo, Pavia, Italy; 4
Amyloidosis Research and Treatment Center, Fondazione IRCCS
Policlinico San Matteo and University o, Pavia, Italy; 5National and Kapodistrian University of Athens, School of Medicine, Greece
Growth differentiation factor-15 (GDF-15), a member of TGFbeta family, is involved in several pathological conditions, including inflammation, cancer, cardiovascular and renal diseases. Serum GDF-15 levels adds prognostic information to conventional prognostic factors, such as NT-proBNP and troponins, in cardiovascular disorders and has been associated with risk of end stage renal disease in diabetics. We evaluated the prognostic significance of serum GDF-15 in two independent cohorts from the Pavia Amyloidosis
e40
-
16th International Myeloma Workshop March 1-4, 2017
Center and the Department of Clinical Therapeutics, Athens. Serum levels of GDF-15 were measured by a novel pre-commercial immunoassay (Roche Diagnostics) in stored serum. The Pavia cohort included 202 and the Athens cohort included 107 patients with AL amyloidosis. Median age and involved FLC levels were similar between the two cohorts but there were differences in other baseline characteristics including heart involvement (77% vs 62% for Pavia vs Athens, p¼0.007), Mayo stage 3(42% in Pavia vs 31% in Athens cohort, p¼0.04), but stage 3B was similar (22% vs 24%) and peripheral nerve involvement (11% in Pavia vs 25% in Athens cohort, p¼0.001). Renal involvement (67% vs 72%, p¼0.415), median eGFR and renal stage distribution were similar (p¼0.544). Two year survival was 59% for Pavia and 56% for Athens cohort. Median GDF-15 levels was 3027 pg/ml in Pavia and 3854 pg/ml in Athens cohort (p¼0.09). The upper quartile of GDF-15 levels, however, was 5658 pg/ml in the Pavia and 7553 pg/ml in Athens cohort, and 90% and 94% of patients in the two cohorts had GDF-15 levels >1200 pg/ml (the upper limit of normal for individuals without cardiovascular disease). We evaluated the prognostic significance of GDF-15 levels for survival in the two cohorts by applying the previously identified cutoff of 7575 pg/ml. GDF-15 above cutoff was associated with shorter survival both in Pavia (17 months vs not reached, p¼0.003) and Athens cohort (13 vs 47 months, p¼0.03). In separate multivariate models for each cohort, that included Mayo stage, GDF-15 remained of independent prognostic significance over Mayo stage, with a hazard ratio (HR) of 1.9 (95% CI 1.2-3.3, p¼0.01) in Pavia cohort and a HR of 1.8 (95% CI 1.2-3.6, p¼0.03) in the Athens cohort. We then evaluated GDF-15 levels for prognosis of renal outcomes (dialysis): GDF-15 >4000 pg/ml was associated with a HR of 7 (95% CI 2015.6, p¼0.001) in Athens cohort (progression to dialysis within 2 years in 7% vs 47%); and HR of 4.9 in Pavia cohort. In both cohorts GDF-15 levels were prognostic for dialysis independent renal stage. Thus, we validated and confirmed in two independent cohorts, with differences in their characteristics, the prognostic value of GDF-15, as a biomarker with prognostic implications for different outcomes in patients with AL amyloidosis. Importantly, GDF-15 emerges also as new biomarkers for renal outcomes in patients with AL amyloidosis.
Indirubins: A Potential Therapeutic Target in Multiple Myeloma Tina Bagratuni,1 Nicolas Gaboriad-Kolar,2 Roubini Zakopoulou,2 Vassilios Myrianthopoulos,2 Efstathios Kastritis,3 Evangelos Terpos,3 Emmanuel Mikros,2 Alexios-Leandros Skaltsounis,2 Meletios Athanasios Dimopoulos4 1 National and Kapodistrian University of Athens, School of Medicine, Athens, Greece; 2National and Kapodistrian University of Athens; 3
national and Kapodistrian University of Athens, School of Medicine,
Greece; 4National and Kapodistrian University of Athens, School of Medicine, Athens, Greece
Current drugs in the treatment of Multiple Myeloma (MM) result in cell death via a number of mechanisms including a direct effect on plasma cells as well as alteration in the BM microenvironment.
Abstracts Among the FDA approved kinase inhibitors, few are based on natural scaffolds. 6-bromoindirubin-3’-oxime (6BIO) is a potent kinase inhibitor based on the natural 6-bromoindirubin scaffold. Indirubin and 6-bromoindirubin are two natural products that have found a particular interest in dye chemistry as the main constituent of indigo and Tyrian purple dyes. Recent findings discovered that 6BIO was a promising anti-cancer agent acting on the JAK/STAT signaling pathway mediating cell proliferation. A library containing 2000 natural molecules was constructed using several data platforms. Each molecule was processed through different filters such as tautomeric studies, protonation and steroisomerism status. Two approaches were followed: the structure-based virtual screening and the ligand-based virtual screening. The results of both approaches were combined, the molecules ranked, and 100 out of 2000 were identified as strong potential bioactive hits for the b5 subunit. Out of these 100 molecules, the chemical structures of high interest were the following: indole alkaloids derivatives (indirubins), flavonoids, secoiridoids, simple phenolic acids and acetophenone. Fifty indirubins derivatives were selected based on different criteria: structure, known/unknown targets, chemodiversity in substitution patterns. To explore the inhibitory effects of indirubins in MM, we performed the WST1 proliferation assay in three MM cell lines (H929, JJN3, L363). Initially, all the selected indirubins(w50 indirubins) were tested at 7.5mM in L363 cell line and proliferation results from the WST1 assay extracted after 24 hours of treatment. More than half of the indirubins tested displayed more than 50% reduction of the proliferation at 7.5mM. Interestingly, 10 out of the 50 indirubinstested reduced more than 80% proliferation after 24 hours. The most active indirubins were tested in H929 and JJN3 cell lines, where similar effects were seen after 24 hours of treatment. All tested indirubins acted in a dose-dependent manner. Among the most active indirubins, two molecules namely 805 and 673 emerged as attractive for further development. Compound 805 is an analog of MLS-2384 while compound 673 is an analog of MLS-2438. The latter derivative represents a promising candidate displaying an IC50 below the micromolar range on H929 and JJN3 cells. To determine the kind of cell death caused by one of the most active indirubins, 673, cell cycle analysis was performed before and after treatment in H929 cell line. In particular changes in RNA expression of 84 genes key to cell cycle regulation were analyzed in H929 cell line. Our results show that among other genes, the ones which have a dramatic increase in their expression.
Primary plasma cell leukemia (pPCL) exhibits poor outcome. We and others have previously demonstrated that novel agents and mainly bortezomib-based regimens (BBR) improve response rates and survival, however the prognostic impact of current treatments and biological markers on the outcome of pPCL has not been sufficiently explored, outside clinical trials. We analyzed the medical records of 50 patients with pPCL (M/F: 25:25; median age 65.5 years, range: 32-86 years; IgG: 19, IgA: 9, light-chain: 14, IgD: 2, non-secretory: 6; ISS1: 5, ISS2: 16, ISS3: 29) out of 2711 myeloma patients (1.8%), registered in the Greek Myeloma study group database, between 2000-2015; 52% of patients had Eastern Cooperative Group (ECOG) performance status 2; 53% had abnormal lactate dehydrogenase (LDH), 28% had hypercalcemia, 68% had hemoglobin <10 g/dL and 65% had high risk cytogenetics; 49/50 patients received therapy: 38 patients received BBR, one patient received melphalan-prednisone-thalidomide and 10 patients received conventional chemotherapy (C/T); 15 patients underwent ASCT consolidation; 48 patients were evaluated for response: 79% achieved objective response (PR) and 35% had at least very good partial response (vgPR), including 17% complete responses. Achievement of vgPR correlated with BBR+ASCT (p¼0.02). After a median follow up of 61 months (95% CI: 34.587.4), 38 patients have died (disease progression: 18, infection: 16, other causes: 4). Early mortality (1 month) occurred in 3/38 deceased patients; 31/38 patients who responded to treatment progressed and 27/31 received 2nd line treatment (lenalidomidebased: 7, BBR: 16, C/T: 4). Progression-free survival was 12 months (95% CI: 8.5-15.4) and it was marginally longer in patients treated with BBR+ASCT vs. others (18 vs. 10 months, p¼0.07). Median OS was 17 months (95% CI: 13-21 months) and it was double in patients treated with BBR+ASCT compared to others (33 months vs. 16 months); median survival after PCL progression was 7 months (95% CI: 3-11 months).
PS-071 Exome Sequencing of AL Amyloidosis Reveals Recurrently Mutated Genes Zuzana Kufova,1 Tereza Sevcikova,2 Petr Vojta,3 Jana Filipova,4 Katerina Growkova,5 Sebastian Grosicki,6 Fedor Kryukov,5 Roman Hajek7 1
Univesity Hospital Ostrava, Ostrava-Poruba, Czech Republic;
PS-070
2
Real-World Data on Clinical Characteristics, Prognosis and Outcome of Primary Plasma Cell Leukemia: A Study of the Greek Myeloma Study Group in the Era of Novel Agents 1
2
Faculty of Medicine, University of Ostrava, Ostrava, Ostrava Zábr eh, Czech Republic; 3Institute of Molecular and Translational Medicine, Palacky University in Olomouc, Olomouc; 4Department of Haemato-Oncology, Univesity Hospital Ostrava, Ostrava-Poruba, Czech Republic; 5Faculty of Medicine, University of Ostrava, Ostrava, Ostrava - Zábr eh; 6Silesian Medical University, Katowice, Poland; 7
University Hospital Ostrava and Faculty of Medicine Ostrava,
Eirini Katodritou, Evangelos Terpos, Sossana Delimpasi,3 Maria Kotsopoulou4
Ostrava, Czech Republic
1
Background: Immunoglobulin light chain amyloidosis (ALA) is a plasma cell dyscrasia characterized by deposition of amyloid fibrils in various organs and tissues, leading to organ dysfunction. Until now, only one systematic study of genomic profile of ALA on exome
Theagenion Cancer Hospital, Thessaloniki, Greece; 2National and
Kapodistrian University of Athens, School of Medicine, Greece; 3
Evangelismos Hospital, Athens, Greece; 4Metaxa Cancer Hospital,
PIRAEUS, Greece
16th International Myeloma Workshop March 1-4, 2017
- e41