ALBERT

Description: Introduction: As previously reported, the combination of brentuximab vedotin with doxorubicin, vinblastine and dacarbazine (A+AVD) demonstrated a statistically significant improvement in modified progression free survival (modified PFS) compared with doxorubicin, bleomycin, vinblastine and dacarbazine (ABVD) in patients with newly diagnosed Stage III or IV classical HL in the phase 3 ECHELON-1 trial (NCT01712490). The benefit of A+AVD in the ITT population observed in the primary analysis was maintained at 3-years median follow-up [3-year PFS: A+AVD: 83.1% (79.9-85.9), ABVD: 76% (72.4-79.2)] and appears independent of interim PET status, disease stage, and prognostic risk factors. Here we present the efficacy and safety results of longer follow-up at a median 43.3 months. Methods: Newly diagnosed patients with Stage III or IV cHL were randomized 1:1 to receive A+AVD (n=664) or ABVD (n=670) intravenously on days 1 and 15 of each 28-day cycle for up to 6 cycles. The primary endpoint of the study was modified PFS per independent central review. The present follow-up PFS analysis is exploratory and per investigator assessment, with a cutoff date of June 17th, 2019. Patients with ongoing peripheral neuropathy (PN) at end of treatment were followed for resolution or improvement (defined as improved by ≥1 grade from worst grade as of the latest assessment) during the post-treatment follow-up period. Results: With a median follow-up of 43.3 months, the 42-month PFS per investigator for all patients was 82.4% (95% CI, 79.1-85.2) on the A+AVD arm and 76.2% (95% CI, 72.6-79.4) on the ABVD arm [overall HR 0.697 (95% CI, 0.547-0.890)]. Exploratory subgroup analyses of PET2(+) and PET2(-) patients showed a treatment effect in favor of A+AVD. The 42-month PFS in PET2(-) patients was 85.0% (95% CI, 81.6-87.7) for A+AVD and 79.6% (95% CI, 75.9-82.8) for ABVD [overall HR 0.695 (95% CI, 0.526-0.919)]; in PET2(+) patients 42-month PFS was 68.3% (95% CI, 54.5-78.7) for A+AVD and 51.5% (95% CI, 38.2-63.4) for ABVD [overall HR 0.552 (95% CI, 0.308-0.989)]. Upon continued follow-up, 81% (356/442) of patients with PN in the A+AVD arm had either complete resolution (64%, 283/442) or improvement (17%, 73/442) of their PN events compared with 83% (236/286) with either complete resolution (74%, 212/286) or improvement (8%, 24/286) in the ABVD arm. Among patients with ongoing PN after continued follow-up, the majority were Grade 1/2 events, with 89% (141/159; 59% Grade 1) and 95% (70/74; 65% Grade 1) on the A+AVD and ABVD arms, respectively. Overall survival data are not yet mature; per protocol, the final analysis will be performed after 112 deaths have occurred. Additional follow-up at an estimated median of ~4 years, including data from prespecified subgroups, will be presented. Conclusions: With a median follow-up of 43.3 months, A+AVD continues to provide a robust, durable benefit for patients with previously untreated Stage III or IV cHL compared with ABVD; the benefit is evident regardless of patient status at interim PET [PET2(+) or PET2(-)] and without the need for treatment intensification. PN continued to completely resolve or improve in patients on the A+AVD and ABVD arms. Together, these data further support the clinical advantages of A+AVD versus ABVD as treatment for patients with previously untreated Stage III or IV cHL. Disclosures Bartlett: Affimed Therapeutics: Research Funding; Bristol-Myers Squibb: Research Funding; Celgene: Research Funding; Dynavax: Research Funding; Forty-Seven: Research Funding; Genentech: Research Funding; Gilead: Research Funding; Immune Design: Research Funding; Janssen: Research Funding; Kite Pharma: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Medimmune: Research Funding; Merck: Research Funding; Millennium: Research Funding; Novartis: Research Funding; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Pharmacyclics: Research Funding; Seattle Genetics, Inc.: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding. Straus:Elsevier (PracticeUpdate): Consultancy, Honoraria; Hope Funds for Cancer Research: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Consultancy, Honoraria. Dlugosz-Danecka:Servier: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Macrogenomics: Research Funding; Roche: Honoraria, Speakers Bureau; Janssen: Consultancy, Honoraria, Speakers Bureau. Illes:Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Seattle Genetics: Research Funding; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees; Roche: Honoraria, Membership on an entity's Board of Directors or advisory committees; Pfizer: Honoraria, Membership on an entity's Board of Directors or advisory committees. Feldman:Takeda: Honoraria, Speakers Bureau; Celgene: Honoraria, Research Funding, Speakers Bureau; Cell Medica: Research Funding; Amgen: Research Funding; Viracta: Research Funding; Bayer: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Kite Pharma: Honoraria, Other: Travel expenses, Speakers Bureau; Janssen: Honoraria, Speakers Bureau; Pharmacyclics: Honoraria, Other: Travel expenses, Speakers Bureau; Pfizer: Research Funding; Portola Pharma: Research Funding; Roche: Research Funding; Eisai: Research Funding; Corvus: Research Funding; Roche: Research Funding; AbbVie: Honoraria, Other: Travel expenses, Speakers Bureau; Seattle Genetics: Consultancy, Honoraria, Other: Travel expenses, Speakers Bureau; Kyowa Hakko Kirin: Research Funding; Trillium: Research Funding. Smolewski:Roche: Other: Travel Expenses. Savage:Seattle Genetics, Inc.: Consultancy, Honoraria, Research Funding; BMS, Merck, Novartis, Verastem, Abbvie, Servier, and Seattle Genetics: Consultancy, Honoraria. Walewski:Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel Expenses, Research Funding; Takeda: Honoraria, Research Funding; Incyte: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Servier: Honoraria; Gilead: Other: Travel Expenses; Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Consultancy, Membership on an entity's Board of Directors or advisory committees; GlaxoSmithKline: Research Funding; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Zinzani:Portola: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Immune Design: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Sandoz: Membership on an entity's Board of Directors or advisory committees; Servier: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; BMS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen-Cilag: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Gilead: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celltrion: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Sanofi: Consultancy; Eusapharma: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; MSD: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Verastem: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Roche: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Kyowa Kirin: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; TG Therapeutics: Honoraria, Speakers Bureau. Hutchings:Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel Expenses, Research Funding; Genmab: Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees, Other: Travel Expenses, Research Funding; Novartis: Research Funding; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel Expenses, Research Funding. Radford:AstraZeneca: Equity Ownership, Research Funding; Novartis: Consultancy, Honoraria; Seattle Genetics: Consultancy, Honoraria; ADC Therapeutics: Consultancy, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; BMS: Consultancy, Honoraria; GSK: Equity Ownership. Munoz:AstraZeneca: Speakers Bureau; Kite Pharma: Consultancy, Research Funding, Speakers Bureau; Pharmacyclics LLC an AbbVie Company: Consultancy, Research Funding, Speakers Bureau; Gilead: Consultancy, Speakers Bureau; Fosunkite: Speakers Bureau; Kyowa: Consultancy, Honoraria, Speakers Bureau; Bayer: Consultancy, Speakers Bureau; Seattle Genetics: Consultancy, Honoraria, Research Funding; Celgene: Research Funding; Portola: Research Funding; Incyte: Research Funding. Kim:Roche: Research Funding; Kyowa-Kirin: Research Funding; Novartis: Research Funding; J&J: Research Funding; Mundipharma: Research Funding; Celltrion: Research Funding; Donga: Research Funding. Advani:Cell Medica, Ltd: Consultancy; Pharmacyclics: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Merck: Research Funding; Kura: Research Funding; Infinity Pharma: Research Funding; Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees, Research Funding; Bayer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene: Research Funding; Celmed: Consultancy, Membership on an entity's Board of Directors or advisory committees; Autolus: Consultancy, Membership on an entity's Board of Directors or advisory committees; Agensys: Research Funding; Stanford University: Employment, Equity Ownership; Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Consultancy, Research Funding; Kyowa Kirin Pharmaceutical Developments, Inc.: Consultancy; Regeneron: Research Funding; Millennium: Research Funding; Janssen: Research Funding; Roche/Genentech: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Gilead Sciences, Inc./Kite Pharma, Inc.: Consultancy, Membership on an entity's Board of Directors or advisory committees; Forty-Seven: Research Funding; AstraZeneca: Consultancy, Membership on an entity's Board of Directors or advisory committees. Ansell:Mayo Clinic Rochester: Employment; Affimed: Research Funding; Mayo Clinic Rochester: Employment; Trillium: Research Funding; Affimed: Research Funding; Seattle Genetics: Research Funding; Trillium: Research Funding; Trillium: Research Funding; Affimed: Research Funding; Bristol-Myers Squibb: Research Funding; Bristol-Myers Squibb: Research Funding; LAM Therapeutics: Research Funding; Regeneron: Research Funding; Affimed: Research Funding; Trillium: Research Funding; Mayo Clinic Rochester: Employment; Regeneron: Research Funding; Bristol-Myers Squibb: Research Funding; Bristol-Myers Squibb: Research Funding; Mayo Clinic Rochester: Employment; LAM Therapeutics: Research Funding; LAM Therapeutics: Research Funding; Seattle Genetics: Research Funding; Mayo Clinic Rochester: Employment; Mayo Clinic Rochester: Employment; Trillium: Research Funding; LAM Therapeutics: Research Funding; Seattle Genetics: Research Funding; Seattle Genetics: Research Funding; Seattle Genetics: Research Funding; Trillium: Research Funding; Affimed: Research Funding; Bristol-Myers Squibb: Research Funding; Seattle Genetics: Research Funding; LAM Therapeutics: Research Funding; Bristol-Myers Squibb: Research Funding; LAM Therapeutics: Research Funding; Regeneron: Research Funding; Regeneron: Research Funding; Seattle Genetics: Research Funding; Regeneron: Research Funding; Trillium: Research Funding; Affimed: Research Funding; Regeneron: Research Funding; LAM Therapeutics: Research Funding; Affimed: Research Funding; Trillium: Research Funding; Affimed: Research Funding; Mayo Clinic Rochester: Employment; Regeneron: Research Funding; Bristol-Myers Squibb: Research Funding; Bristol-Myers Squibb: Research Funding; Mayo Clinic Rochester: Employment; Regeneron: Research Funding; LAM Therapeutics: Research Funding; Affimed: Research Funding; Regeneron: Research Funding; LAM Therapeutics: Research Funding; Seattle Genetics: Research Funding; Seattle Genetics: Research Funding; Bristol-Myers Squibb: Research Funding; Trillium: Research Funding; Mayo Clinic Rochester: Employment. Younes:Roche: Consultancy, Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Curis: Honoraria, Research Funding; Merck: Honoraria, Research Funding; Abbvie: Honoraria; Takeda: Honoraria; Pharmacyclics: Research Funding; AstraZeneca: Research Funding; Genentech: Research Funding; Biopath: Consultancy; Xynomics: Consultancy; Epizyme: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; HCM: Consultancy; BMS: Research Funding; Syndax: Research Funding. Gallamini:Takeda: Consultancy; Roche: Consultancy. Miao:Millennium Pharmaceuticals, Inc., Cambridge, MA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Employment, Equity Ownership. Liu:Takeda: Employment. Fenton:Seattle Genetics, Inc.: Employment, Equity Ownership. Forero-Torres:Seattle Genetics: Employment, Equity Ownership, Honoraria, Research Funding.

Description: Background: The mTOR kinase inhibitor, rapamycin (RAPA), inhibits cell growth by G1 phase cell cycle arrest. RAPA-induced cytotoxicity against acute lymphoblastic leukemia (ALL) cells was recently reported. Aim: We investigated cytotoxicity of RAPA alone or in combination with fludarabine (FA), a cytostatic active in G1 phase, in ALL cells as well as in healthy lymphocytes. Methods: NALM-6 cells (pre-B ALL-derived cell line), and phytohemaglutynin (PHA)-stimulated normal lymphocytes were treated for 24–48h with different concentrations of RAPA (0,05–500ng/ml), alone or in combination with 0.1–50.0mM FA. Some samples were initially incubated with RAPA for 24h and then FA was added for the next 24h. Untreated cultures and treated with RAPA, FA or PHA alone served as respective controls. The pro-apoptotic effect was assessed by both Annexin-V and TUNEL assays and presented as an apoptotic index (AI). Cell cycle was analyzed by DNA content distribution in propydium iodide/RN-ase stained cells. Additionally, intracellular expression of cyclin A, D3 and E was evaluated. All fluorescence measurements were performed by flow cytometry. Overall cytotoxicity was evaluated by MTT assay. Results: RAPA was found to exert dual effect on NALM-6 cells. In lower concentrations (0.05–5ng/ml) RAPA exclusively inhibited proliferation, arresting NALM-6 cells in G1 phase of cell cycle. An increasing evidence of apoptosis, along to enhancing cytotoxicity in MTT assay was observed higher RAPA doses (10–500ng/ml). When NALM-6 were treated with RAPA+FA, the highest AIs were found for the combination of 0.5 or 1.0ng/ml RAPA with 1nM FA. Median AI induced at 24h by 1.0ng/ml RAPA+FA was 12.4%, comparing to with RAPA or FA alone (AIs 1.9% and 4.8%, respectively; both p

Description: Background: Bortezomib (Velcade, PS-341), a selective inhibitor of the 26S proteasome which was recently approved for the treatment of patients with refractory multiple myeloma, showed also anti-leukemic activity in several studies. Bortezomid alone was found to induce apoptosis of chronic lymphocytic leukemia (CLL) cells in vitro. So far, there was no data on effects of its combination with other drugs routinely used in this disease. Aim: Cytotoxic activity of bortezomib in combination with purine nucleoside analogues, fludarabine (FA) and cladribine (2-CdA) was tested in CLL cells in vitro. Methods: The study was performed on CLL lymphocytes derived from 26 patients. Cell viability was assessed by propidium iodide staining. Apoptosis was evaluated by both Annexin-V and caspase activation flow cytometry assays. Additionally, expression of the apoptosis-regulating proteins Bax, Bak, Bid, Bcl-w, Bcl-2, XIAP and Mcl-1 was evaluated in CLL cells. Results: Bortezomib alone induced significant, dose-dependent cytotoxicity, inducing apoptosis of CLL cells. The viability of CLL cells treated with bortezomib decreased from 73.4%±14.9 (at 1.25nM) to 39.1%±15.4% (at 10nM). The mean apoptotic index estimated after 24h of treatment was in a range from 29.9%±18.0% at the dose of 1.25nM to 66.1%±21.8% at 10nM of bortezomib. Combination of this agent with FA or 2-CdA resulted in an increased cytotoxicity, comparing single drugs. It was especially evident when 5nM of bortezomib was combined with suboptimal doses of FA or 2-CdA. Cytotoxic effect of 5nM bortezomib +FA was significantly higher than 5nM bortezomib (p=0.0004) or FA (p

Description: Combination of vincristine, doxorubicin and dexamethasone (VAD regimen) is a widely-used, effective and economic initial treatment of multiple myeloma (MM). However, between 30 and 50% of MM patients do not respond to VAD, and exposure to VAD may induce multi-drug resistance. Therefore, in the era of emerging novel therapies for MM, there is a need for pre-treatment identification of potential responders to VAD. Importantly, all three VAD components have common steps in their pathways of transport and metabolism, thus even modest inherited alteration caused by single nucleotide polymorphisms (SNP) may influence VAD efficacy an toxicity. The aim of this study was to investigate whether analysis of common functional SNP in drug transporter P-glycoprotein gene (MDR1 or ABCB1), phase I cytochrome P450A metabolizing enzymes genes (CYP3A4 and CYP3A5), phase II glutathione S-transferase P metabolizing enzyme gene (GSTP1) as well as in glucocorticoid receptor gene NR3C1 can be useful to predict response to VAD in MM. Fifty-two uniformly treated MM patients were investigated for 8 common SNPs including MDR1 exon 12 C1236T, MDR1 exon 21 G2677T/A, MDR1 exon 26 C3435T, CYP3A4*1B, CYP3A5*3, GSTP1 A313G as well as NR3C1 bcl I and NR3C1 N363S polymorphisms. Genotypes were identified using direct sequencing or RFLP methods. The response to VAD was estimated after administration of 3 courses of VAD, and patients who achieved complete or partial remission of MM were assigned as responders to VAD. In the cohort of included MM patients there were 39/52 (75%) responders to VAD. The analysis of the prognostic impact of the genotyped SNPs showed that carriers of P-glycoprotein gene MDR1 2677G allele had a significantly greater probability of achievement of the response to VAD as compared to the non-carriers, odds ratio(OR)=8.0, 95% confidence interval (95%CI)= 1.2–52.9, Fisher exact test p=0.031). Moreover, the predictive effect of MDR1 2677G allele was further increased by the presence of glucocorticoid receptor NR3C1 gene 363N allele. The carriers of both MDR1 2677G and NR3C1 363N alleles had 8.4-fold greater probability to achieve complete or partial remission with 3 courses of VAD (OR=8.40, 95% CI=1.8–40.0, p=0.008). Taken together, these data support the hypothesis that analysis of common functional polymorphisms in drug transporters, metabolizing enzymes and receptors may be useful for individualization of the initial therapy of MM. Particularly MDR1 G2677T/A and NR3C1 N363S SNPs can be considered as determinants of response to VAD therapy.

Description: Despite of evident progress in treatment of chronic lymphocytic leukemia (CLL), the disease still remains incurable. Several attempts have been made therefore to develop most effective and selective therapeutical approaches. A promising approach that involves targeting RNA either by the use of specific antisense oligonucleotides or cytostatic/cytotoxic ribonucleases is recently being promoted. Two such ribonucleases, onconase (ONC; ranpirnase) and R-amphinase (R-Amph), derived from Rana pipiens oocytes, have been developed. ONCdemonstrated preferential toxicity to tumor cells and was shown to be effective in vivo in animal tests as well as in clinical trials in treatment of malignant mesothelioma. Moreover, ONC is synergistic when used in combination with a variety of antitumor modalities including anthracyclines. R-Amph was developed only recently and thus far there is only a single report demonstrating its cytostatic and cytotoxic activity against human promyelocytic HL-60-, Jurkat T-cell- and U-937 histiomonocytic leukemic cells in vitro. In the present study we aimed to assess potential cytotoxicity of ONCand R-Amph against CLL cells. Toward this aim, leukemic cells were isolated from 36 untreated patients with CLL and were cultured for 24–72 h with either ONC or R-Amph alone and in combination with purine analogues, cladribine (2-CdA) and fludarabine (FA), two drugs routinely used in treatment of CLL, as well as with doxorubicin (DOX), the drug reported to show synergy with ONC in solid tumors. Cytotoxicity of the study drugs was assessed by the propidium iodide exclusion assay using flow cytometry. Their pro-apoptotic activity was examined by the Annexin-V (Ann-V) binding test, detection of caspase-3, -8, and -9 activation, a decrease of mitochondrial potential and the expression of apoptosis–regulating proteins from the Bcl-2 family. Compensated apoptotic index (CAI) has been calculated based on Ann-V assay as a difference in the percentage of apoptotic cells between the drug-treated sample and spontaneous apoptosis in the parallel untreated control. After preliminary experiments the optimal concentrations of both ONC and R-Amph were found to be 20 μg/ml; these were the lowest doses that induced significant cytotoxicity during 24–72 h of incubation, in comparison with parallel controls. The significant effect of ONC was evident after 48 h of treatment (median CAI=11.5%; p=0.035 versus control). After 72 h of incubation the median CAI for ONC was 17.1% (p=0.009). The significant cytotoxicity of R-Amph was seen after 72 h incubation (median CAI =19.9%; p=0.007, respectively). The mechanism of this cytotoxicity involved the induction of apoptosis along its mitochondrial pathway, with the drop of mitochondrial potential and activation of caspase-9 and caspase-3, concurrent with an increase in expression of pro-apoptotic Bax protein (p=0.035 versus control; after 72h) and a decrease of anti-apoptotic Bcl-2 expression (p=0.006; after 72 h). No significant changes in expression of Bak and Mcl-1 were observed. Synergistic effect was found for both, ONC plus 2-CDA and ONC plus FA (combination indices, CI;

Description: Abstract 2520 Objectives: Notch-1 signaling pathway is considered important for acute myeloid leukemia (AML) pathogenesis. Delta-like 1 (DLL-1) and Jagged-1 are functional opposite members of Notch-1 ligand family. There is evidence that Jagged-1 has oncogenic potential whereas DLL-1 is tumor suppressive in AML biology. Moreover, it has been shown that Jagged-1 can promote AML development by stimulation of Notch-1 receptor and in independent manner. The mechanism of DLL-1 function in AML remains unknown. This study was conducted to elucidate the clinical role of Notch-1 and its ligands in AML patients. The expression of Jagged-1, DLL-1 and Notch-1 was estimated in leukemic blasts of newly diagnosed AML patients. Additionally, the level of examined proteins was correlated with known prognostic factors, response to treatment, overall survival (OS) and relapse free survival (RFS). Methods: The Jagged-1, DLL-1 and Notch-1 expression was assessed in leukemic blasts obtained from bone marrow or peripheral blood of 75 AML patients with median age 58 years (21–82y). The expression of proteins was estimated by using multi-color flow cytometry. The isotype controls were performed for all measurements. The percentage of Notch-1, Jagged-1 and DLL-1 positive cells served as protein expression. Based on the median cut-off value patients were subdivided into low-expressers and high-expressers group. Results: The median (Me) expression of Notch-1 and its ligands Jagged-1 and DLL-1 was 2,2% (range 0,1–24,8%); 20,6% (range 0,9–89,7%) and 3% (range 0,1–35,7%); respectively. Significant correlation between Jagged-1 and Notch-1 protein expression in AML blasts (r=0,24; p=0.048) as well as a trend toward a positive correlation between Jagged-1 and DLL-1 expression (r=0,36; p=0.059) were observed. The highest expression of Notch-1 and DLL-1 but not Jagged-1 was observed in patients with poor-risk karyotype according to SWOG classification (Me 7,9%; Me 7,65%) as compared to good-risk (Me 1,1%, p

Description: Abstract 2842 Background: Several B-cell-derived malignancies still remains incurable. A promising approach that involves targeting RNA either by the use of specific antisense oligonucleotides or cytostatic/cytotoxic ribonucleases (RNases) is being promoted. Two amphibian RNases, onconase (ONC; ranpirnase) and, more recently, R-Amphinase (r-Amph), have already been developed, but studied so far mainly in solid tumors. In this report we demonstrate ex vivo and in vivo antitumor activity of ONC and, for the first time, R-Amph against tumor cells from different B-cell lymphoid malignancies. Material and methods: Pro-apoptotic effects of ONC or r-Amph alone and in combinations with doxorubicin (DOX) or purine analoques, fludarabine (FA) and cladribine (2-chlodeoxyadenozine; 2-CDA) were assessed ex vivo in peripheral blood samples of untreated patients with acute lymphoblastic leukemia; ALL, N=15) and chronic lymphocytic leukemia (CLL, N=45). Lymphocytes obtained from 31 healthy volunteers was also treated. Additionally, we tested in vitro activity of both agents alone or in combination with doxorubicin (DOX) on cell lines derived from Burkitt lymphoma (Raji) and with DOX, vincristin (VCR), dexamethasone (DEX), mafosfamide (MAF) or anti-CD20 antibody, rituximab (RIT) on diffuse large B-cell lymphoma (DLBCL)-derived cells (Toledo). Moreover, multiple myeloma (MM) cells were treated in vitro (RPMI 8226) and ex vivo (bone marrow cells). Drug-induced apoptosis was assessed after 48–72 hr incubation by the flow cytometry Annexin-V assay. Additionally, a drop of mitochondrial potential, activation of caspases -8, -9 and -3, and expression of several apoptosis-regulating proteins were investigated. Compensated apoptotic index (CAI) was calculated as a difference in % of apoptotic cells between the drug-treated sample and the parallel untreated culture. Results: Based on preliminary studies on different ex vivo and in vitro experimental models, concentrations 10–20 mg/ml of ONC and 20–60 mg/ml of r-Amph were chosen for final studies as the lowest doses inducing significant pro-apoptotic effect. In similar manner concentrations 50 ng/ml of 2-CdA, 1mg/ml of FA, 10–20 mg/ml of DOX, 20 mg/ml of RIT, 1ng/ml of VCR, 20 mg/ml of DEX and 1 mg/ml of MAF were selected. In CLL cells, significant effect of both ONC and r-Amph was evident after 72 hrs of treatment (median CAI 17.1% and 13.9%; vs untreated control cells p=0.009 and 0.012, respectively). Importantly, the same doses did not affect survival of healthy lymphocytes. Combination of ONC, but not r-Amph, with 2-CdA or FA exerted significantly stronger effect than single drugs (p

Description: An optimal treatment for patients with primary refractory or relapsed (R/R) Hodgkin lymphoma (HL) who did not respond to standard salvage therapy has not been established. Brentuximab vedotin (BV) and bendamustine (B) used as monotherapy were shown to be active in R/R HL and both are recommended as a therapeutic option. It has been proposed that the combination of BV with B (BVB) may improve the outcome of R/R HL patients. To test this hypothesis, we compared retrospectively the efficacy and safety of BV and BVB regimen in R/R HL patients. Methods: Since March 2014 all patients with R/R HL treated at PLRG centers were offered either BV in monotherapy every 21 days or in combination with B (B 90 mg/m² on days 1 and 2 of treatment cycle) according to the policy in each center. Results: BV or BVB therapy was administered to 94 patients (median age 33 years, range 18-68) with primary refractory (n=54) or relapsed HL (n=40), including 34 patients after autologous stem cell transplant (ASCT). The patients were treated with the median of 3 (range, 2-12) prior chemotherapy lines. Fifty-seven patients received BV and 37 patients BVB regimen. In 16 of them, BVB was de-escalated to BV after the median of 2 cycles (range, 2-7). The patients' and disease characteristics did not differ between two groups. In the whole study group, the median number of applied cycles per patient was 4 (range, 2-16). Dose-limiting toxicities were observed in 8% of patients. No difference was found between BV and BVB group, with similar rate of grade 3-4 neutropenia (16% vs 13%) and thrombocytopenia (4% vs 3%). Lung infection occurred in 1 patient treated with BV (2%) and 3 treated with BVB (2% vs 8%, p ns), with 2 treatment-related deaths in the BVB group. The response rate after 2 cycles of BV and BVB treatment defined as complete (CMR) or partial metabolic response (PMR) assessed by positron emission tomography was 69% (26% of CMR and 43% of PMR) and 89% (46% of CMR and 43% of PMR), respectively (p=0.036). After 4-6 cycles of treatment, CMR, PMR and stable metabolic disease was documented in 59%, 19% and 13% in the BV group vs. 71%, 17% and 3%, respectively, in the BVB group (p ns). Finally, 36 patients (22 from BV and 14 from BVB group) proceeded to ASCT, and 17 (9 from BV and 8 from BVB group) to allogeneic SCT. The overall and progression-free survival at 24 months were 80% and 51%. The survival curves did not differ between two study groups. In conclusion, our results suggest that BVB is a feasible regimen that provides higher response rate after 2 cycles compared to BV monotherapy. However, the overall response rate achieved after 4-6 cycles of treatment are comparable between BV monotherapy and combined BVB treatment. Infectious complications, including serious lung infections, were observed in both study groups, with two treatment-related deaths in the BVB group. Disclosures Dlugosz-Danecka: Roche: Consultancy; Servier: Consultancy. Jurczak:Nordic Nanovector: Research Funding; Epizyme: Research Funding; Beigene: Research Funding; Bayer: Research Funding; Celgene: Research Funding; Merck: Research Funding; Servier: Consultancy, Honoraria, Research Funding; European Medicines Agency: Consultancy; AstraZeneca: Consultancy; Gilead: Consultancy, Research Funding; Afimed: Research Funding; Morphosys: Research Funding; Janssen: Consultancy, Research Funding; Sandoz-Novartis: Consultancy; Roche: Research Funding; Pharmacyclics: Research Funding; Acerta: Consultancy, Research Funding; TG therapeutics: Research Funding. Zaucha:Roche: Honoraria; Takeda: Honoraria; Amgen: Honoraria; Celgene: Honoraria.