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The mutational status of the variable region of the immunoglobulin heavy chain (IGHV) genes remains the most significant prognostic factor in chronic lymphocytic leukemia (CLL) patients. However, the groups of mutated (M) and unmutated (UM) patients are also heterogeneous, and additional markers are used for a more accurate prognosis. The aim of our work was to determine the prognostic value of the signs of antigen selection determined by BASELINe statistics in M IGHV sequences of CLL patients. Clinical data, IGHV gene configuration, TP53, NOTCH1, SF3B1 mutations were analyzed in 127 CLL patients with M IGHV sequences. The median OS of patients with negative selection in the framework regions (FWRs) of IGHV genes was 120 months compared to 202 month in other CLL patients (P = 0.016). In multivariate Cox regression analysis Binet stage C vs A + B (P 0.0001), SF3B1 mutations (P 0.0001), negative selection in the FWRs (HR P = 0.007), and age ≥65 years (P = 0.034) were powerful adverse prognostic factors for OS in CLL patients with M IGHV genes. These preliminary data suggest that the signs of antigen-driven selection may be used as a prognostic factor in CLL patients with M IGHV genes in combination with other markers.

Summary Background In the LYM-3002 study, the efficacy and safety of frontline bortezomib plus rituximab, cyclophosphamide, doxorubicin, and prednisone (VR-CAP) and rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) were compared in transplant-ineligible patients with untreated, newly diagnosed, mantle cell lymphoma. We report the final overall survival and safety outcomes for patients in the long-term follow-up phase after the primary progression-free-survival endpoint was met. Methods LYM-3002 was a randomised, open-label, phase 3 study done at 128 clinical centres in 28 countries in Asia, Europe, North America, and South America. Adult patients with confirmed stage II–IV previously untreated mantle cell lymphoma, Eastern Cooperative Oncology Group performance status score of 2 or less, who were ineligible for bone marrow transplantation, were randomly assigned (1:1) to receive six or eight 21-day cycles of VR-CAP (intravenous rituximab 375 mg/m2, cyclophosphamide 750 mg/m2, doxorubicin 50 mg/m2, and bortezomib 1·3 mg/m2, plus oral prednisone 100 mg/m2) or R-CHOP (intravenous vincristine 1·4 mg/m2 [2 mg maximum], rituximab 375 mg/m2, cyclophosphamide 750 mg/m2, and doxorubicin 50 mg/m2, plus oral prednisone 100 mg/m2). Randomisation was done according to a computer-generated randomisation schedule prepared by the sponsor; permuted blocks central randomisation was used (block size of 4), and was stratified by International Prognostic Index score and disease stage at diagnosis. The primary endpoint of this final analysis was overall survival, which was analysed in the intention-to-treat population. This study is registered with ClinicalTrials.gov , number NCT00722137 , and is closed to new participants with follow-up completed. Findings Between May 22, 2008, and Dec 5, 2011, 487 patients were enrolled and randomly assigned. 268 patients (140 in the VR-CAP group and 128 in the R-CHOP group) were included in the follow-up analysis, which included patients with data available after the primary analysis clinical cutoff date of Dec 2, 2013. After median follow-up of 82·0 months (IQR 74·1–94·2), median overall survival was significantly longer in the VR-CAP group than in the R-CHOP group (90·7 months [95% CI 71·4 to not estimable] vs 55·7 months [47·2 to 68·9]; hazard ratio 0·66 [95% CI 0·51–0·85]; p=0·001). Three new adverse events were reported since the primary analysis cutoff (one each of grade 4 lung adenocarcinoma and grade 4 gastric cancer in the VR-CAP group, and one case of grade 2 pneumonia in the R-CHOP group). 103 (42%) of 243 patients in the VR-CAP group, and 138 (57%) of 244 in the R-CHOP group died; the most common cause of death was progressive disease. Interpretations Compared with R-CHOP, VR-CAP was associated with significantly longer survival, and had a manageable and expected safety profile. Our results support further assessment of VR-CAP in patients with previously untreated mantle cell lymphoma. Funding Janssen Research & Development.

Objectives To analyze the distribution of single nucleotide polymorphisms (SNPs) in the TP53 gene in chronic lymphocytic leukemia (CLL) patients and to evaluate their associations with clinical behavior of the disease. Methods SNPs in exons and parts of adjacent introns of the TP53 gene were analyzed in 235 CLL patients observed during 2005-2012 years. Data on individuals of European descent from the 1000 Genomes Project data set were used as a reference. Results In the recessive model of inheritance, we found borderline associations between CLL risk and C/C genotype of rs1642785 (p = 0.048); G/G genotype of rs2909430 (in men only; p = 0.036) and Pro72Pro genotype of rs1042522 (in men only; p = 0.045). Risk of CLL was increased also in carriers of rare haplotypes (p = 0.0049). Besides, genotypes Pro72Pro of rs1042522, C/C of rs1642785, and G/G of rs2909430 were associated with an increased incidence of TP53 mutations. Median of overall survival in rs1800372 carriers was comparable to that of patients with TP53 mutations. Other evaluated SNPs were not associated with survival. Conclusion Our data suggest that some TP53 variants may affect the risk of CLL. rs1800372 polymorphism might be the marker of unfavorable prognosis of the disease.

Background: Acalabrutinib is a highly selective, potent inhibitor of Bruton tyrosine kinase, an essential part of the B-cell signalling pathway. Methods: ASCEND is a global, multicentre, randomised, open-label, Phase 3 study. Eligible patients, aged ≥18 years with relapsed/refractory chronic lymphocytic leukemia (CLL), were randomised 1:1 centrally via an interactive voice/web response system, and stratified by del(17p) status, Eastern Cooperative Oncology Group performance status, and prior lines of therapy, to receive oral acalabrutinib 100 mg twice daily (BID) or investigator's choice (ie, idelalisib BID plus rituximab ["I-R"] or bendamustine plus rituximab ["B-R"]). The primary endpoint was progression-free survival (PFS) assessed by an independent review committee in the intention-to-treat population. Safety was assessed in all patients who received any treatment. Findings: From February 21, 2017 to January 17, 2018, 310 patients were randomised to acalabrutinib (n=155) or investigator's choice therapy (n=155 [I-R, n=119; B-R, n=36). Patients had received a median of two prior therapies (range 1-10). After a median follow-up of 16·1 months (interquartile range 14·0-18·2), median PFS was significantly longer with acalabrutinib (not reached [95% CI 14·9-not reached]) compared with investigator's choice (16·5 months [95% CI 14·0-17·1]; HR 0·31 [95% CI 0·20-0·49]; p

8501 Background: Selinexor is an oral, selective inhibitor of XPO1-mediated nuclear export, leading to the reactivation of tumor suppressor proteins. In a phase 1b/2 study, the combination of once weekly (QW) selinexor with bortezomib and dexamethasone (SVd) was well tolerated with anti-MM activity in patients (pts) with PI-sensitive and PI-refractory disease. While twice weekly (BIW) bortezomib in combination therapy is efficacious, prolonged use is limited due to peripheral neuropathy (PN, 50-60%). The BOSTON study was designed to determine if SVd improves progression free survival (PFS), overall response rates (ORR) and reduces the rate of PN vs Vd. Methods: BOSTON is a global, phase 3, randomized study of QW SVd vs BIW Vd after 1-3 prior anti-MM regimens. The primary endpoint is PFS. Secondary endpoints include ORR, overall survival (OS) and PN (rates and EORTC QLQ-CIPN20 outcomes). Randomization is stratified by treatment with prior PI therapies, number of prior anti-MM regimens (1 vs > 1), and Revised International Staging System (R-ISS; Stage III vs I or II). Following confirmation of progressive disease, pts on Vd could cross over to either: 1) SVd for pts able to tolerate continued bortezomib or 2) selinexor and dexamethasone for pts with bortezomib intolerance. Results:402 pts were enrolled; 195 and 207 to SVd and Vd, respectively. Median age was 67 (range: 38-90). Most (59.6%) pts were > 65 years and 57.1% were male. R-ISS stage at the time of MM diagnosis was III for 18.5% of pts. Baseline characteristics were balanced across the 2 arms. SVd significantly prolonged PFS vs Vd (median 13.93 vs 9.46 months, HR = 0.70, P = 0.0066). SVd was associated with a significantly higher ORR (76.4% vs 62.3%, P = 0.0012). Median OS was not reached on SVd vs 25 months on Vd (P = 0.28). Most frequent treatment-related adverse events (grade ≥3) for SVd vs Vd were thrombocytopenia (35.9% vs 15.2%), fatigue (11.3% vs 0.5%) and nausea (7.7% vs 0%). Clinically important differences were reported on the motor, autonomic and sensory scales on CIPN20. PN rates (grade ≥2) were significantly lower with SVd vs Vd (21.0% vs 34.3%, P = 0.0013). Conclusions: BOSTON is the first phase 3 study to evaluate the clinical benefit of SVd for relapsed/refractory MM. The study met the primary endpoint: once weekly SVd significantly improved PFS and ORR compared to twice weekly Vd. Rates of PN were significantly reduced with numerically fewer deaths on SVd vs Vd. Full dataset will be presented at the meeting. Clinical trial information: NCT03110562 .

8005 Background: PD-1 blockade via pembro monotherapy showed antitumor activity in R/R cHL. KEYNOTE-204 (NCT02684292) was a randomized, international, open-label, phase III study of pembro vs BV in R/R cHL. Methods: Patients (pts) were aged ≥18 y, were post−autologous stem cell transplant (auto-SCT) or ineligible for auto-SCT, and had measurable disease and ECOG PS 0 or 1. BV-naive and BV-exposed pts were eligible. Pts were randomized 1:1 to pembro 200 mg IV Q3W or BV 1.8 mg/kg IV Q3W and stratified by prior auto-SCT (yes vs no) and status after 1L therapy (primary refractory vs relapsed

© 2018 The Authors., [Background]: The present prospective, multinational, noninterventional study aimed to document and describe real-world treatment regimens and disease progression in multiple myeloma (MM) patients. [Patients and Methods]: Adult patients initiating any new MM therapy from October 2010 to October 2012 were eligible. A multistage patient/site recruitment model was applied to minimize the selection bias; enrollment was stratified by country, region, and practice type. The patient medical and disease features, treatment history, and remission status were recorded at baseline, and prospective data on treatment, efficacy, and safety were collected electronically every 3 months., [Results]: A total of 2358 patients were enrolled. Of these patients, 775 and 1583 did and did not undergo stem cell transplantation (SCT) at any time during treatment, respectively. Of the patients in the SCT and non-SCT groups, 49%, 21%, 14%, and 15% and 57%, 20%, 12% and 10% were enrolled at treatment line 1, 2, 3, and ≥ 4, respectively. In the SCT and non-SCT groups, 45% and 54% of the patients had received bortezomib-based therapy without thalidomide/lenalidomide, 12% and 18% had received thalidomide/lenalidomide-based therapy without bortezomib, and 30% and 4% had received bortezomib plus thalidomide/lenalidomide-based therapy as frontline treatment, respectively. The corresponding proportions of SCT and non-SCT patients in lines 2, 3, and ≥ 4 were 45% and 37%, 30% and 37%, and 12% and 3%, 33% and 27%, 35% and 32%, and 8% and 2%, and 27% and 27%, 27% and 23%, and 6% and 4%, respectively. In the SCT and non-SCT patients, the overall response rate was 86% to 97% and 64% to 85% in line 1, 74% to 78% and 59% to 68% in line 2, 55% to 83% and 48% to 60% in line 3, and 49% to 65% and 36% and 45% in line 4, respectively, for regimens that included bortezomib and/or thalidomide/lenalidomide., [Conclusion]: The results of our prospective study have revealed great diversity in the treatment regimens used to manage MM in real-life practice. This diversity was linked to factors such as novel agent accessibility and evolving treatment recommendations. Our results provide insight into associated clinical benefits., Writing support during the development of our report was provided by Laura Mulcahy and Catherine Crookes of FireKite, an Ashfield company, a part of UDG Healthcare plc, which was funded by Millennium Pharmaceuticals, Inc, and Janssen Global Services, LLC. The EMMOS study was supported by research funding from Janssen Pharmaceutical NV and Millennium Pharmaceuticals, Inc.

Chronic lymphocytic leukemia (CLL) is an incurable disease. Quality of life during treatment and periods of subsequent remission is therefore vital. Health-related quality of life (HRQoL) was compared in relapsed CLL during and after treatment with ofatumumab combined with fludarabine and cyclophosphamide versus fludarabine and cyclophosphamide alone. The European Organisation for Research and Treatment of Cancer (EORTC) QLQ-C30 v3 and QLQ-CLL16 were used to assess HRQoL in this open-label, phase 3 study. Improvements in prespecified domains of patient-reported outcomes (Global Health Status [GHS]/HRQoL and B symptom scores) were recorded in both treatment arms after three cycles and were sustained after 18 months of follow-up. The two treatment arms were not significantly different at the nominal 0.05 level for GHS/HRQoL (p = .7278) or B symptoms (p = .5968). Small improvements in quality of life were maintained after therapy. The addition of ofatumumab was without any adverse impact on HRQoL (NC...

In this multicenter, open-label, phase III study, patients with relapsed chronic lymphocytic leukemia (CLL) were randomized (1:1) to receive ofatumumab plus fludarabine and cyclophosphamide (OFA + FC) or FC alone; the primary endpoint being progression-free survival (PFS) assessed by an independent review committee (IRC). Between March 2009 and January 2012, 365 patients were randomized (OFA + FC: n = 183; FC: n = 182). Median IRC-assessed PFS was 28.9 months with OFA + FC versus 18.8 months with FC (hazard ratio = 0.67; 95% confidence interval, 0.51–0.88; p = .0032). Grade ≥3 adverse events (≤60 days after last dose) were reported in 134 (74%) OFA + FC-treated patients compared with 123 (69%) FC-treated patients. Of these, neutropenia was the most common (89 [49%] vs. 64 [36%]). OFA + FC improved PFS with manageable safety for patients with relapsed CLL compared with FC alone, thus providing an alternative treatment option for patients with relapsed CLL. Trial registration: www.clinicaltrials.gov...

We report results of a randomized, phase III study of ofatumumab versus physicians' choice treatment in patients with bulky fludarabine-refractory chronic lymphocytic leukemia and explore extended versus standard-length ofatumumab treatment. Patients (79 ofatumumab, 43 physicians' choice) completed a median 6 (ofatumumab) or 3 (physicians' choice) months' therapy. Ofatumumab-treated patients with stable disease or better were randomized (2:1) to 6 months' extended ofatumumab treatment or observation. Although the study did not meet the primary endpoint of progression-free survival (PFS) by independent review committee (ofatumumab: 5.4 months, physicians' choice: 3.6 months; p = 0.27), median PFS by investigators was significantly longer for ofatumumab versus physicians' choice (7.0 versus 4.5 months; p = 0.003) as was time to next therapy (median 11.5 versus 6.5 months; p = 0.0004). PFS and time to next therapy were significantly longer with ofatumumab extended treatment than observation (p = 0.026 and p = 0.002, respectively; n = 37). The adverse-event profile of long-term ofatumumab administration showed no unexpected findings (Clinicaltrials.gov identifier: NCT01313689).

Clinical data and immunoglobulin variable heavy chain (IgVH) gene configuration were analyzed in 47 CLL patients, exposed to ionizing radiation (IR) due to Chernobyl NPP accident, and 141 non-exposed patients. Clean-up workers of the second quarter of 1986 (n=19) were picked out as separate group with the highest number of unmutated cases (94.4%), increased usage of IgVH1-69 (33.3%) and IgVH3-21 (16.7%) genes, high frequency of secondary solid tumors (6 cases) and Richter transformation (4 cases). These preliminary data suggest that CLL in the most suffered contingent due to Chernobyl NPP accident might have some specific features.

Summary Background Remarkable advance in the knowledge of the biology and treatment of chronic lymphocytic leukaemia (CLL) has been noted in recent years. CLL resembles several heterogeneous diseases with variable morphological features. New agents with antileukemic activity have been introduced recently for CLL patients , which improved the treatment results. Fludarabin has shown its efficacy in CLL both as a treatment of choice and in combination with conventional cytostatic agents. Clinical and haematological features of B-CLL in persons affected by the Chernobyl accident in the last period are presented. Aim The aim of this study was the comparison of two chemotherapy different schemes for B-CLL treatment. Materials/Methods The study was based on 32 patients with diagnosis of B-CLL who were treated at the Hematology Department. Depending upon the protocol all patients were classified in two subgroups: I – fludarabine monotherapy treatment, II – fludarabine with combination with other chemotherapy preparations. Results Of the treatment are analyzed for 16 patients after different chemotherapy regimens. A combined fludarabine and cyclophosphamide regimen has been shown to improve long-term complete and partial remission in the majority of patients as compared with a more conservative approach.

rs6449182 CD38 gene polymorphism was determined by polymerase chain reaction with restriction of products in 328 chronic lymphocytic leukemia (CLL) patients and 271 age- and sex-matched controls. An association between GG genotype and CLL risk was found in the whole group of patients (OR=2.12; p=0.009) and in patients with unmutated immunoglobulin heavy chain variable genes (OR=2.17; p=0.011) comparing to the controls. In the subgroup of 174 controls with evaluated lipids the genotype distributions in CLL patients and dyslipidemic controls were similar. An association between GG genotype and CLL risk was significant compared to controls without lipids' abnormalities (OR=3.92; p=0.006).

BCD-020 (Acellbia, rituximab biosimilar candidate) was shown to be highly similar to innovator rituximab (MabThera®/Rituxan®) in terms of its quality characteristics, in vitro biological activity, as well as toxicology and PK/PD characteristics in non-human primates. International multicenter comparative randomized open-label clinical study was carried out in a period from 2011 to 2013 and involved over 30 centers in Russia, Ukraine and India. Its methodology and design complies with current EMA guidelines on similar biological products containing monoclonal antibodies (EMA/CHMP/BMWP/403543/2010). 92 patients with follicular non-Hodgkin’s lymphoma, stage I-IV by Ann Arbor, or marginal zone lymphoma, stage I-IV by Ann Arbor, ECOG 0-2, who had at least 1 measurable lesion were enrolled. According to study protocol patients with secondary transformed B-cell lymphomas or with highly aggressive types of tumor, bulky disease, severe concomitant somatic disorders and some other conditions were excluded. If a patient had previous story of chemotherapy or radiation he could be included after at least 3 weeks post-treatment. Participation of patients who were previously treated with any kind of monoclonal antibodies was prohibited. After signing standard informed consent form and completion of 28-days screening period eligible patients underwent stratification in accordance to their prognostic risk (FLIPI or IPI) and previous treatment (naïve or pretreated). Subsequently patients were randomized (1:1) into 2 groups: 46 patients were included in the main group where Acellbia (rituximab biosimilar) was administered at a dose of 375 mg/m2 as a slow IV infusion on day 1, 8, 15 and 22; 46 patients were included in the reference group where MabThera was used at the same regimen. Use of any other medicines for the treatment of lymphoma was strictly prohibited. Efficacy was assessed on the basis of computed tomography and bone marrow evaluation which were performed 1 month after the completion of treatment. Median age of patients in each group was 57.5 years (main group [50.0-64.0], reference group [47.0-65.0]). Manageable comorbidities were reported in 50% of patients in the main group and 34.78% of patients in the reference group, p=0.2053. Comparative analysis of the prognostic risk factors confirmed the equivalence of study groups. The number of pretreated patients in both groups was equal – 8 individuals per group. Statistical analysis didn’t find any difference in overall response rate in general population of patients (39.52% patients in the main group vs. 36.57% patients in the reference group, p=0.8250), as well as in population of pretreated patients (28.6% vs 37.5% respectively, p=1.00) and in population of naïve patients (42.8% vs 39.4% respectively, p=1.00). The lower limit of the two-tailed 95% CI for difference in proportions of ORR was equal to -0.17 and exceeded the predefined non-inferiority margin -0.2, which confirmed non-inferiority of Acellbia to MabThera in terms of efficacy. Treatment-associated AE of any grade were reported in 21.74% patients in both arms, in the absence of statistically or clinically significant difference (p = 0.8005). There were 2 cases of CTCAE 4.03 grade 3-4 AEs in each group. PK and PD parameters were shown to be equivalent in both study groups. Thus, study results suggest that Acellbia has same efficacy and safety in patients with B-cell non-Hodgkin’s lymphoma. Disclosures Chernyaeva: JCS BIOCAD: Employment. Ivanov:JCS BIOCAD: Employment. Isaev:JCS BIOCAD: Employment.

OFA was approved for the treatment of fludarabine and alemtuzumab refractory CLL on the basis of a single-arm study which also included pts with BFR CLL. This open label, randomized, Phase III study of OFA or PC treatment in pts with CLL refractory to fludarabine and with bulky lymphadenopathy (at least one lymph node > 5 cm) was conducted to confirm the results of OFA in BFR pts and also through a 2nd randomization to compare extended OFA (12 months) vs. approved regimen of OFA (6 months). PC was considered an appropriate comparator for these pts given the lack of a consensus around standard of care treatment for this difficult-to-treat population. Pts were randomized 2:1 to receive either the approved regimen of 8 weekly infusions of OFA followed by 4 monthly infusions (Dose 1, 300 mg; Doses 2–12, 2000 mg) or PC of non-OFA containing therapy, including treatments approved for CLL and well established standards of care for up to 6 months. Pts in the OFA arm who had an investigator-assessed CR, PR or SD underwent a 2nd 2:1 randomization to 6 additional OFA infusions 2000 mg every 4 weeks (OFA Ext), or observation (Obs). Pts in the PC arm who developed disease progression during the study could receive OFA salvage therapy for up to 12 months of treatment. Premedication for OFA infusions consisted of glucocorticoid, paracetamol, and antihistamine. The primary objective of this study was to evaluate progression-free survival (PFS) with OFA treatment versus PC as assessed by an Independent Review Committee (IRC). Response assessments were performed using the 2008 Update of the NCI-WG CLL Guidelines [Hallek, 2008]. Secondary endpoints included overall response rate (ORR), time to next therapy (TNT), overall survival (OS), and safety. 122 pts were randomized (79 OFA, 43 PC). Pts completed a median of 12 OFA infusions and 3 months PC therapy. The median PFS (mPFS) as measured by IRC was 5.4 months for OFA and 3.6 months for PC (hazard ratio [HR] 0.79, p=0.27, stratified log rank test; Fig 1). Median PFS as assessed by the investigator (INV) was 7.0 months for OFA and 4.5 months for PC (HR 0.56, p=0.003). The median time to start of next anti-CLL treatment was 11.5 months for OFA and 6.5 months for PC (p=0.0004, stratified log rank test). The ORR (95% CI) by IRC evaluation was 38% (27%, 50%) for OFA and 16% (7%, 31%) for PC (p=0.02). 37 pts underwent a 2nd randomization to either OFA Ext (n=24) or Obs (n=13). From 2nd randomization, mPFS (INV) was 5.6 months for OFA Ext and 3.5 months for Obs (HR: 0.49, p=0.026, stratified log rank test) (Fig 2). Grade ³3 infusion reactions occurred in 5% of pts in the OFA arm with no fatal reactions in either arm. Grade ≥3 cytopenias in the OFA arm included neutropenia (24%, no prolonged neutropenia and 1 patient with late onset neutropenia), thrombocytopenia (8%), anemia (8%). Of the 43 PC pts, 22 received OFA Salvage with an ORR of 50% and mPFS of 5.4 months. Table of Response and Time to event Endpoints OFA (N=79) PC (N=43) Characteristics Median (range) Age, years 61.5 (46 – 82) 63 (40 – 76) No. prior therapies 4 (2 – 16) 3 (2 – 11) No. of infusions 12 (1 – 18) 3 (1 – 6) IRC Investigator Efficacy endpoints OFA PC p-value OFA PC p-value Responders, % 38% 16% 0.019 49% 37% 0.415 mPFS, months 5.36 3.61 0.267 7 4.5 0.003 OFA PC TNT, months 11.5 6.5 p=0.0004 OS, months 19.2 14.5 p=0.13 Figure 1. Kaplan-Meier Estimates of PFS by IRC (OFA vs. PC) Figure 1. Kaplan-Meier Estimates of PFS by IRC (OFA vs. PC) Figure 2. Kaplan-Meier Estimates of PFS by INV (OFA Ext vs. Obs from 2nd randomization) Figure 2. Kaplan-Meier Estimates of PFS by INV (OFA Ext vs. Obs from 2nd randomization) Conclusions: Although the study did not meet its primary endpoint of demonstrating statistically significant superior PFS by IRC, the ORR, PFS by investigator, TNT, and OS favor OFA and were consistent with previously reported results in BFR pts. There was also a longer PFS in pts who underwent the 2nd randomization to receive OFA Ext vs. the approved OFA treatment regimen of 6 months treatment. Disclosures Österborg: GSK: Research Funding. Off Label Use: Discussion of extended ofatumumab dosing (12 months) in addition to the approved 6 month dosing of ofatumumab in refractory CLL. Zaritskey:Novartis: Consultancy, Honoraria. Kaplan:GSK: Honoraria. Steurer:GSK: Consultancy, Honoraria, Research Funding. Schuh:GSK: Honoraria; Roche: Honoraria; Gilead: Honoraria; Celgene: Honoraria; Napp: Honoraria. Montillo:GSK: Honoraria. Kulyaba:GSK: Honoraria. Gorczyca:GSK: Employment. Daly:GSK: Employment. Chai-Ni:GSK: Employment. Lisby:Genmab: Employment. Gupta:GSK: Employment.

The leukemia cells of unrelated patients with chronic lymphocytic leukemia (CLL) display a restricted repertoire of immunoglobulin gene rearrangements and very similar structure of B-cell receptor (stereotyped BCR). Numerous different subsets of stereotyped BCRs were identified. Some associations between specific BCRs subsets, clinical course of CLL, and molecular abnormalities were found. Namely, short overall survival, accumulation of SF3B1 mutations in subset #2, high risk for Richter’s transformation, trisomy 12, NOTCH1 mutations in subset #8, increased frequency of NOTCH1 mutations in subset #1 were revealed (Rossi et al., 2009, 2013; Streffoed et al., 2013). The aim of this study was to evaluate distribution of different stereotyped BCR subsets in Russian and Ukrainian CLL cohort with special attention on prognostic unfavorable subsets. Methods Immunoglobulin heavy chain variable (IGHV) genes of 969 CLL patients (564 from Russia and 405 from Ukraine) were amplified according to BIOMED-2 rules, sequenced and analyzed with IMGT and IgBlast databases. Additionally, NOTCH1 c.7544_7545delCT mutations were analyzed in 301 patients (randomly collected and in the prognostic unfavorable subsets) by amplification refractory mutation system (ARMS) PCR as proposed by Rossi D. et al., 2012. In 19 cases NOTCH1 mutational status was confirmed by direct DNA Sanger sequencing. Results Joined Russian-Ukrainian CLL cohort was presented mainly by unmutated (UM) IGHV gene cases (64.2%). The distribution of major clusters of stereotyped BCR did not differ from those presented in the literature (comparison was made separately for cases with mutated (M) and UM IGHV genes, taking into account the differences in the mutational status of IGHV genes between our CLL cohort and CLL cohorts from European countries and USA), except for subset #8, which was found in one case only (0.16% of UM IGHV gene cases). At the same time, subset #8 (IGHV4-39/IGHD6-13/IGHJ5 rearrangement) belongs to the group of “major” subsets in CLL (Agathangelidis et al., 2012), and its frequency varies from 0.9% to 2% among cases with UM IGHV genes in the most of presented CLL cohorts (Murray et al., 2008; Messmer et al., 2009; Maura et al., 2011). Frequency of subset #2 had tendency to be lower among Russian CLL patients (1.4% of M CLL cases), but not among Ukrainian patients (4.5% of M CLL cases) comparing with European CLL cohorts. NOTCH1 c.7544_7545delCT was identified in 27 out of 301 patients (9.0%), which was in accordance with other CLL studies. Twenty five cases with NOTCH1 mutations had UM IGHV genes and 2 cases expressed M IGHV3-21 gene (both subset #2). As well, majority of NOTCH1 mutated cases (17/27, 63%) showed stereotyped BCRs, which belonged to the following subsets: #1 (2 of 12), #2 (2 of 7), #3 (1 of 6), #7 (2 of 10), #9 (2 of 10), #25 (1 of 2), #59 (1 of 3) (subset’s designation according Murray et al., 2008), UA/ref2 (1 of 2), UA/ref11 (2 of 2) (subset’s designation according Bilous et al., 2010), or having a homology with the CLL sequences of the public CLL database. No NOTCH1 mutations were found in 11 IGHV4-39-used cases, including one case from subset #8. In 10 of 27 (37.0%) NOTCH1 mutated cases IgHV1-69 gene was used, while among wild type NOTCH1 patients IgHV1-69 was presented in 65/274 (23.7%) cases (p=0.12). Conclusion The distribution of the major subsets of stereotyped BCRs in CLL patients from Russia and Ukraine (eastern slavic countries) was similar to the western European countries and USA, except for low frequency of subset #8, associated with unfavorable clinical course. Our preliminary data suggested that NOTCH1 c.7544_7545delCT mutation is typical for UM CLL cases with stereotyped BCRs, but no clear association with specific subsets was found in the absence of subset #8. Disclosures: No relevant conflicts of interest to declare.

Abstract 4580 Chronic lymphocytic leukemia (CLL) was shown to be the most common leukemia among nuclear workers and persons, who were exposed to ionizing radiation (IR) due to Chernobyl NPP accident (Romanenko A. et al., 2008). Some clinical features were also indicated, such as: younger age, more advanced symptoms at the onset of disease, more aggressive course with rapid progression, poor response to standard chemotherapy (Klymenko V. et al., 1999, Kryachok I. et al., 2005). The p53 signaling pathway plays an important role in the biology of CLL. Inactivating alterations of TP53 gene by mutation or deletion is strongly associated with adverse clinical outcome and drug resistance. Some single nucleotide polymorphisms (SNP), in particular SNP within TP53 gene at codon 72 (Arg/Pro) and p21 at codon 31 (Ser/Arg) were recently reported may affect p53 pathway function, and influence clinical outcome of CLL. In our study we focused on investigation of these functional SNPs. TP53 SNP at codon 72 and p21SNP at codon 31 were investigated in 79 IR exposed CLL patients, 76 IR non-exposed CLL patients and 72 controls. CLL patients and control group were compared by gender and age, and two of CLL groups were also compared by clinical data at diagnosis. Genotyping was done using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method according to Hiroshi H. et al., 2007. The digested products were separated on a 3% agarose gel and analyzed. The both of studied SNPs were in Hardy-Weinberg equilibrium in all groups. The distribution of TP53 genotypes in IR-exposed CLL patients (Arg/Arg 49.4%; Arg/Pro 38%; Pro/Pro 12.6%) did not differ significantly from the distribution in IR non-exposed CLL cohorts (Arg/Arg 44.7%; Arg/Pro 46.1%; Pro/Pro 9.2%; p=0.549) and control group (Arg/Arg 59.9%; Arg/Pro 33.3%; Pro/Pro 6.9%; p=0.201). There were higher frequency of p21 Ser/Arg genotype in IR-exposed CLL patients (Ser/Arg 21.1%; Ser/Ser 78.9%) compared to IR non-exposed CLL (Ser/Arg 8.1%; Ser/Ser 91.9%; p=0.025), but not to controls (Ser/Arg 13.9%; Ser/Ser 86.1%; p=0.453). We analyzed the association of studied genotypes with some characteristics of CLL patients such as: clinical data at the time of diagnosis (stage, initial white blood cell count, size of lymph nodes), time-to-treatment (TTT), response to treatment, overall survival (OS), progression-free survival (PFS), development of autoimmune complications, secondary tumors, Richter transformation, and immunoglobulin heavy chain variable (IGHV) gene mutation status. Some associations were revealed. Namely, patients with p21 Ser/Arg genotype compared to patients with Ser/Ser genotype had: significantly shorter TTT period (5 mo vs 18 mo; p=0.039), especially in IGHV mutated group (1 mo vs 12 mo; p=0.022), tendency to bulky disease (13.6% vs 5.5%; p=0.16), B-symptoms at diagnosis (22.7% vs 11.8%; p=0.116) and lower frequency of secondary tumors development (4.5% vs 20.5%; p=0.073). Among patients with unmutated IGHV genes TP53 Pro/Pro carriers had shorter PFS (35 mo vs 108 mo; p=0.05) compared to Arg/Arg and Arg/Pro carriers. In summary, we revealed increased number of p21 codon 31 Ser/Arg carriers among IR-exposed CLL patients in comparison with IR non-exposed CLL patients. Further studies are required to evaluate possible role of p21 SNP in development of CLL under IR influence. Disclosures: No relevant conflicts of interest to declare.

Aim The aim of our research was an evaluation of possibility to use PRAME gene expression as a marker of minimal residual disease (MRD) in multiple myeloma (MM) patients. Previously we have detected high frequency of PRAME gene expression among patients with advanced stages of MM (68.5%). Material and Methods Two pairs of primers complementary to PRAME exon 5 and 6 were designed. PRAME gene expression was analyzed in bone marrow cells by 2-step reverse-transcription-polymerase chain reaction (RT-PCR). 24 initially PRAME-posistive MM patients were studied on day +100 and one year after high dose therapy with autologous peripheral blood stem cell transplantation (PBSCT). Results On day +100 after PBSCT PRAME expression was found in 22 cases, in 5 patients in the absence of plasma cells in bone marrow. After 1 year among 5 patients with complete clinico-hematological remission PRAME positive cells persisted in 3 cases. Patients who were PRAME-negative on day +100 are still in remission. Conclusion Detection of PRAME gene seems to be used as a molecular marker for definition of MRD. Our results suggest that complete molecular remission after PBSCT may be achieved only in a few MM patients.