Search

Adult T-cell leukemia-lymphoma (ATL) has been divided into 4 clinical subtypes: acute, lymphoma, chronic, and smoldering. The aim of this study is to develop a novel prognostic index (PI) for chronic and smoldering ATL. We conducted a nationwide retrospective survey on ATL patients, and 248 fully eligible individuals were used in this analysis. In the univariate analysis, sex, performance status, log10 (soluble interleukin-2 receptor [sIL-2R]), neutrophils count, and lymphadenopathy showed values of P < .05 in training samples. A multivariate analysis was performed on these factors, and only log10 (sIL-2R) was identified as an independent prognostic factor in training samples. Using a regression coefficient of this variable, a prognostic model was formulated to identify different levels of risk: indolent ATL-PI (iATL-PI) = 1.51 × log10 (sIL-2R [U/mL]). The values calculated by iATL-PI were divided into 3 groups using a quartile point. In the validation sample, median survival times (MSTs) were 1.6 years, 5.5 years, and not reached for patients in the high-, intermediate-, and low-risk groups, respectively (P < .0001). To make the scoring system clinically practicable, we simplified iATL-PI according to trichotomizing sIL-2R at 1000 and 6000 U/mL, using a quartile point. Patients with more than 6000 U/mL sIL-2R were categorized into the high-risk group, less than and equal to 1000 U/mL into the low-risk group, and the others into the intermediate-risk group, and MSTs were 1.6 years, not reached, and 5.5 years, respectively (P < .0001). iATL-PI has potential as a novel tool for a risk-adapted therapeutic approach.

We assessed long-term safety and efficacy of CT-P10 and rituximab in patients with newly diagnosed low-tumour-burden follicular lymphoma (LTBFL), and following a single transition from rituximab to CT-P10. This double-blind, parallel-group, active-controlled phase 3 trial randomized patients with CD20+ LTBFL to receive CT-P10 or US-sourced rituximab (375 mg/m2 intravenous). Induction therapy (weekly for 4 cycles) was followed by a 2-year maintenance period for patients achieving disease control (CR, CRu, PR and SD). During the maintenance, CT-P10 or rituximab were administered every 8 weeks (6 cycles) in the first year and additional CT-P10 was administered every 8 weeks (6 cycles) in the second year. Secondary endpoints (reported here) were overall response rate during the study period, progression-free survival, time-to-progression, and overall survival. Safety and immunogenicity were also evaluated over the study period. Between Nov 9, 2015 and Jan 4, 2018, 258 patients were randomised (130 CT-P10; 128 rituximab). Over the study period, 115 (88%; CT-P10) and 111 (87%; rituximab) patients achieved overall response. At a median follow-up of 29·2 months (IQR: 26·1-33·7), median progression-free survival, time-to-progression, and overall survival were not estimable. The KM estimates (95% CI) for OS at 36 months were 98% (93-99) and 97% (89-99) in the CT-P10 and rituximab groups, respectively. Corresponding values for PFS were 80% (70-87) and 68% (54-79), while results for TTP were 82% (72-88) and 68% (54-79) in the CT-P10 and rituximab groups, respectively. (Figure A. OS; Figure B. PFS and Figure C. TTP) Over the study period, 114 (88%) and 104 (81%) patients in the CT-P10 and rituximab groups, respectively, experienced at least one treatment-emergent adverse event (TEAE) and 14 (11%) patients in each group experienced TE-serious adverse events (TESAEs). There were no unexpected safety findings observed during the second year of the maintenance period after single transition from rituximab to CT-P10. Figure 1 Disclosures Kwak: Celltrion Healthcare: Membership on an entity's Board of Directors or advisory committees; Xeme Biopharma/Theratest: Other: equity; CJ Healthcare: Consultancy; Sellas Life Sciences Grp: Consultancy; Enzychem Life Sciences: Membership on an entity's Board of Directors or advisory committees; Antigenics: Other: equity; InnoLifes, Inc: Consultancy, Membership on an entity's Board of Directors or advisory committees; Pepromene Bio: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celltrion, Inc.: Consultancy. Sancho:Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees; Gilead: Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Honoraria; Gelgene: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: 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. Kim:Alexion Pharmaceuticals Inc.: Honoraria, Research Funding. Menne:Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Kite/Gilead: Honoraria, Speakers Bureau; Novartis: Honoraria, Other: Travel costs, Speakers Bureau; Pfizer: Honoraria, Other: Travel costs, Speakers Bureau; Celgene: Honoraria, Other: Travel grants; Roche: Speakers Bureau; Janssen: Honoraria, Research Funding, Speakers Bureau; Astra Zeneca: Research Funding; Takeda: Honoraria, Speakers Bureau. Jurczak:Maria Sklodowska-Curie National Research Institute of Oncology, Krakow, Poland: Current Employment; Jagiellonian University, Krakow, Poland: Ended employment in the past 24 months; Acerta: Research Funding; Bayer: Research Funding; Janssen: Research Funding; MeiPharma: Research Funding; Pharmacyclics: Research Funding; Roche: Research Funding; Takeda: Research Funding; TG Therapeutics: Research Funding. Trneny:Gilead: Consultancy, Honoraria, Other: Travel Expenses; Janssen: Consultancy, Honoraria, Other: Travel Expenses; Roche: Consultancy, Honoraria, Other: Travel Expenses; MorphoSys: Consultancy, Honoraria; Celgene: Consultancy; Incyte: Consultancy, Honoraria; Takeda: Consultancy, Honoraria, Other: Travel Expenses; Bristol-Myers Squibb Company: Consultancy, Honoraria, Other: Travel Expenses; Amgen: Honoraria; Abbvie: Consultancy, Honoraria, Other: Travel Expenses. Ogura:Cellgene: Honoraria; Chugai: Honoraria; Denovo Biopharma: Membership on an entity's Board of Directors or advisory committees; MejiSeika Pharma: Membership on an entity's Board of Directors or advisory committees; Mundi Pharma: Membership on an entity's Board of Directors or advisory committees; SymBio: Membership on an entity's Board of Directors or advisory committees; TevaTakeda: Membership on an entity's Board of Directors or advisory committees; Verastem: Membership on an entity's Board of Directors or advisory committees; Daiichi Sankyo: Membership on an entity's Board of Directors or advisory committees; Celltrion, Inc.: Honoraria, Membership on an entity's Board of Directors or advisory committees; Eisai: Membership on an entity's Board of Directors or advisory committees. Kim:Pfizer: Research Funding; Donga: Research Funding; Mundipharma: Research Funding; F. Hoffmann-La Roche: Research Funding; Kyowa Kirn: Research Funding; Celltrion: Research Funding; JJ: Research Funding. Lee:Celltrion, Inc.: Current Employment. Kim:Celltrion, Inc.: Current Employment. Ahn:Celltrion, Inc.: Current Employment. Buske:Roche, Janssen, Bayer, MSD: Research Funding; Morphosys: Membership on an entity's Board of Directors or advisory committees; Roche, Janssen, AbbVie, Pfizer, Celltrion: Honoraria, Speakers Bureau. OffLabel Disclosure: Rituximab monotherapy to LTBFL patients

Background: Regulatory T cells (Treg) play important roles in onset of graft-versus-host disease (GVHD) and chronic GVHD (cGVHD) in elderly people is more frequent than in younger people after allogeneic hematopoietic stem cell transplantation (HCT). However, little is known about the differences in the number and function of Treg and natural killer (NK) cells in elderly and young patients. In this study, we examined the cause of the difference in incidences of cGVHD in these patient populations by measuring Treg and NK cells over time after HCT. Patients and Methods: The subjects were 65 consecutive patients aged >18 years who underwent HCT for different hematologic diseases at our hospital between December 2008 and December 2018 and survived for >100 days after HCT. The median age was 57 years (range: 19-73 years), and those aged ≥58 (n=33) and ≤57 (n=32) were classified as elderly and younger patients, respectively. Evaluated variables included recipient and donor characteristics, and transplant-related factors, including conditioning regimen, donor type, degree of match, and GVHD prophylaxis. After obtaining informed consent, blood was drawn from all patients on day 100 and at 12-month intervals thereafter. Analysis of CD4+CD25+Foxp3+ Treg cells, CD3+CD4+ T cells, CD3+CD8+ T cells, CD19+ B cells, and CD3-CD56+ NK cells was performed using flow cytometry, and the absolute numbers of these cells were calculated. NK activity was measured by the standard 51Cr release assay at an effector:target (E:T) ratio of 20:1. Results: The incidence of grades II-IV acute GVHD was 29.2% at 100 days. There was no difference in the incidence of acute GVHD between elderly and younger patients [HR=0.85, 95% CI 0.34-2.09, p=0.72]. However, the overall incidence of cGVHD was 37.1%. In univariate analysis, the incidence of cGVHD in elderly patients was significantly higher than that in younger patients [HR=3.61, 95% CI 1.13-11.59, p=0.031]. Elderly patients had a significantly lower percentage of B cells than younger patients at 24 months (13.4±9.6% vs. 27.0±15.6%, p=0.024) and 36 months (16.4±10.3% vs. 31.1±11.2%, p Conclusion: The percentage and absolute number of Treg cells in elderly patients were significantly lower than those in younger patients, whereas CD8+ T cells were significantly higher in elderly patients after HCT. The absolute number of Treg cells tended to reconstitute one year after HCT in younger patients, but tended to decrease gradually in elderly patients, with no recovery even three years after HCT. In contrast, there was no change in NK cells. Similar results were obtained in an analysis excluding patients who developed cGVHD. These results suggest that a difference in immunological reconstitution between elderly and younger patients may explain the difference in the incidence of cGVHD between these patients. This may be useful in determining the choice of therapy with consideration of the immunologic mechanism. Disclosures Suzumiya: Celgene, Kyowa Kirin, Chugai-Roche, Eisai, Takeda, Celltrion, SymBio, Astellas, Ono, AstraZeneca, Ootsuka, Taiho, Mundi, Dainihon-Sumitomo: Research Funding.

Background: Follicular lymphoma (FL) is the most common indolent B-cell lymphoma, and its clinical behavior is heterogeneous. FL is sensitive to immunochemotherapy with anti-CD20 antibody, but characterized by a high rate of relapse and having a risk of histological transformation. Bendamustine + rituximab (BR) therapy is one of standard treatment options for both untreated and relapsed FL. There are several reports that 18[F]-Fluoro-2-deoxy-D-glucose(¹⁸F-FDG) PET-CT (PET) response after induction therapy can predict progression-free survival (PFS) in FL undergoing first-line immunochemotherapy. However, the significance of PET after second-line therapy has rarely been evaluated, especially in a prospective setting. Thus, we conducted a single arm prospective phase 2 study to compare 1-year (1-yr) PFS between patients with PET-positive and PET-negative at end-of-treatment (EOT) response assessment using standardized PET and CT after the BR therapy in patients with first relapsed or recurrent FL (UMIN000013756). Method: The W-JHS NHL01 study is a prospective multicenter phase 2 trial, which included adult (≥20 years) patients with relapsed or recurrent FL (CD20 positive, grade 1-3a), ECOG-PS (≤2) and measurable lesion (CT minor axis >1.0cm). Patients had to have one prior regimen with chemotherapy and/or rituximab. Histologic transformation at recurrence was ineligible. Patients were treated with 6 cycles of BR (R 375 mg/m2 d1, B 90 mg/m2 d1-2, every 28 days). EOT PET within 1-2 months after last BR was evaluated, and assessments for progression were made using contrast enhanced CT scans at 12 and 18 months after the registration of this study and at physician's discretion thereafter. Standardized PET and contrast enhanced CT were evaluated centrally by independent expert radiologists and the PET positivity was defined by a residual ¹⁸F-FDG uptake score of 4 or greater (5-point scale). The efficacy of treatment was evaluated by CT scan according to Cheson 2007 criteria. The primary endpoint was 1-yr PFS. Key secondary endpoints were overall response rate (ORR), CR rate (CRR), 1-yr overall survival (OS). Results: Between July 4, 2014 and Jan 24, 2017, 75 patients were enrolled, eight of whom were excluded from the analysis. Because 4 received fewer than 4 cycles of BR, 2 received with non-standardized PET and 2 had progression disease. Of the remaining 67 patients in the efficacy-evaluable population, median (range) age was 63 (47-83) and 27/67 (40%) were male. 60% of enrolled patients had received R-CHOP for first-line treatment. 60 (80%) patients received 6 cycles of BR. Among 67 patients, median follow-up of surviving patients was 23.9 months (12.2-45.1). ORR of 67 patients was 97.0% (95%CI: 89.6-99.6%), CRR was 59.7% (95%CI: 47.1-71.5%). 1-yr PFS in 72 patients was 88.9% (95%CI:80.7-94.3%). 1-yr OS in all patients was 97.3% (95%CI:89.6-99.3%). The EOT PET positive rate after BR was 9/67 (13.4%). The prevalence according to 5-point scale was 3% in score 1, 70% in score 2, and 7.5% in score 3. 1-yr PFS was 77.8% [95% confidence interval (CI): 36.5- 93.9%] in EOT PET-positive group (n=9) and 93.1% (95% CI: 82.6-97.3%) in EOT PET-negative group (n=58), with a significantly favorable 1-yr PFS in patients with EOT PET-negative group (log-rank test: p = 0.02) (Figure 1). In addition, there was no statistically significant difference between 1-yr PFS and SUVmax after BR therapy. Standardized PET before BR was done in 29 patients. ΔSUV of ¹⁸F-FDG uptake was not significantly related with 1-yr PFS. And there were no significant relation between 1-yr PFS and FLIPI or FLIPI2. The prevalence of grade 3 and 4 adverse events (AEs) was lymphopenia (77.3%), neutropenia (32%), leukopenia (30.7%), thrombocytopenia (4%), and anemia (1.3%). In non-hematologic AEs, there was each patient with appendicitis, herpes zoster, reactivation of HBV, nausea, general fatigue, acneiform eruption, hypernatremia, hyponatremia and hypocalcemia. Conclusions: BR as a second-line treatment for FL was effective with high ORR of 97% and high 1-yr PFS of 88.9%. EOT PET could predict PFS in patients with relapsed/recurrent FL treated with BR. Disclosures Rai: Chugai: Speakers Bureau. Suzumiya:Celgene, Kyowa Kirin, Chugai-Roche, Eisai, Takeda, Celltrion, SymBio, Astellas, Ono, AstraZeneca, Ootsuka, Taiho, Mundi, Dainihon-Sumitomo: Research Funding. Izutsu:Kyowa Kirin, Eisai, Takeda, MSD, Chugai, Nihon Medi-physics, Janssen, Ono, Abbvie, Dainihon Sumitomo, Bayer, Astra Zeneca, HUYA Japan, Novartis, Bristol-Byers Squibb, Mundi, Otsuka, Daiichi Sankyo, Astellas, Asahi Kasei: Honoraria; Celgene: Consultancy; Eisai, Symbio, Chugai, Zenyaku: Research Funding; Eisai, Chugai, Zenyaku: Honoraria; Chugai, Celgene, Daiichi Sankyo, Astra Zeneca, Eisai, Symbio, Ono, Bayer, Solasia, Zenyaku, Incyte, Novartis, Sanofi, HUYA Bioscience, MSD, Astellas Amgen, Abbvie, ARIAD, Takeda, Pfizer: Research Funding. Nishikori:Eisai, Ono, Sumitomo Dainippon: Research Funding; Bristol Meyers Squibb, Janssen, MSD, Chugai, Eisai, Sumitomo Dainippon, Kyowa Kirin, Mundipharma, Takeda, Nippon Shinyaku, Ono, Pfizer: Honoraria. Shibayama:Celgene, Chugai, Eisai, AstraZeneca: Membership on an entity's Board of Directors or advisory committees; Astellas, Teijin, MSD, Shionogi, Eisai, Sumitomo Dainippon, Taiho, Nippon Shinyaku: Research Funding; Takeda, Novartis, Janssen, Chugai, Eisai, Mundi Pharma, Ono, Otsuka, Kyowa Kirin, Sumitomo Dainippon, AstraZeneca, Avvie, DaiichiSankyo, Fujimoto, Nippon Shinyaku, Sanofi, Bristol-Myers Squibb, Pfizer: Honoraria. Maeda:Mundipharma Co Ltd.: Honoraria; Kyowa Kirin Co. Ltd.: Honoraria; Bristol-Myers Squibb: Honoraria, Research Funding; Astellas Pharma Inc.: Research Funding. Yamauchi:Astellas, AbbVie: Consultancy; Pfizer, Chugai, Teijin, Solasia: Research Funding. Akashi:Celgene, Kyowa Kirin, Astellas, Shionogi, Asahi Kasei, Chugai, Bristol-Myers Squibb: Research Funding; Sumitomo Dainippon, Kyowa Kirin: Consultancy. Kanakura:Chugai, Eisai: Research Funding.

Introduction Aggressive natural killer cell leukemia (ANKL) is a rare leukemic form of mature natural killer cell neoplasms that is closely associated with Epstein-Barr virus. ANKL presents a fulminant clinical course, resulting in a poor prognosis with a median overall survival of approximately 2 months. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is currently the only curative treatment, but the long-term outcomes after allo-HSCT remain unclear. Methods Using the national Japanese transplant registry database, the outcomes of 59 ANKL patients who underwent first allo-HSCT between 1997 and 2016 were analyzed. Correlation between groups was examined by the c2 test, Mann-Whitney U test, and Kruskal-Wallis test. Patient survival data were analyzed using the Kaplan-Meier method and compared by the log-lank test. The cumulative incidence of relapse and non-relapse mortality were calculated considering competing risks. Results The median patient age was 37 years (range, 9 to 66), and males accounted for 68%. The median time from diagnosis to allo-HSCT was 3.7 months (range, 1.1 to 13.9). Twenty-nine patients received stem cells from cord blood (CB), 18 received them from peripheral blood (PB), and 12 received them from bone marrow (BM). Two patients had a prior history of autologous HSCT. Twenty-one patients (36%) had a complete response (CR) and 7 (12%) had a partial response (PR), but 31 (52%) were not responding at the time of allo-HSCT. Forty-four patients received myeloablative conditioning and 15 received non-myeloablative conditioning. Thirty-two patients received tacrolimus-based GVHD prophylaxis, including 1 with additional post-transplant cyclophosphamide as part of haploidentical HSCT, whereas 26 received cyclosporin-based GVHD prophylaxis. The median follow-up of survivors was 62 months (range, 0.9 to 193). The median OS and relapse-free survival were 3.9 months and 2.6 months, respectively. The probability of OS and relapse-free survival, and cumulative incidence of relapse and non-relapse mortality 1 year after HSCT were 33.9%, 32.4%, 55.5%, and 12.1%, respectively. The probability of OS was significantly higher for patients with CR or PR at allo-HSCT than for those without a response (40.6% vs 16.1% at 5 years; P = 0.046). Among the 24 patients with primary induction failure (PIF) at allo-HSCT, 15 achieved CR after allo-HSCT. The prognosis of these 15 patients was almost equivalent to that of those who received allo-HSCT in CR or PR, as shown in the Figure (P = 0.95). Regarding the stem cell source, the probability of OS was significantly higher for patients who received stem cells from CB than for those who received them from PB or BM (CB 37.3% vs PB 15.8% and BM 16.7% at 5 years; P = 0.04). Seventeen patients (59%) of those who received stem cells from CB were CR at HSCT, whereas only 4 patients (13%) of those who received stem cells from PB or BM were CR at HSCT. In addition, 5 (83%) of 6 patients who received stem CB transplantation in PIF achieved CR after allo-SCT, whereas 10 (56%) of 18 patients who received PB stem cell transplantation or BM transplantation in PIF achieved CR. The age and year at HSCT were not different between the groups. The time from diagnosis to allo-HSCT did not differ among stem cell sources (median CB 3.3 months, PB 3.7 months and BM 4.1 months; P = 0.31). Regarding the conditioning regimen, the probability of OS was not different between myeloablative and non-myeloablative conditioning regimens (P = 0.58). Patients who developed acute GVHD grade 1/2 had a significantly better prognosis than those with grade 3/4 or without GVHD (P < 0.001). In contrast, chronic GVHD development did not affect the prognosis (P = 0.60). At the last follow-up, 42 patients (71%) had died. The most common cause of death was primary disease (62%), followed by infection (14%) and organ dysfunction (7%). Conclusion Allo-HSCT can lead to long-term survival even for patients with PIF at HSCT. CB is useful as a stem cell source, providing good outcomes and timely allo-HSCT for this rapidly progressive disease. To confirm our findings and evaluate the outcome of allo-SCT in more detail, further studies including patients who did not receive allo-SCT for ANKL are warranted. Figure Disclosures Ishida: Bristol-Meyers Squibb: Research Funding; Pfizer: Research Funding; Astellas Pharma: Research Funding; Eli Lily and Company: Research Funding; Celgene: Honoraria; Chugai Pharmaceutical: Consultancy, Research Funding. Izutsu:Celgene: Consultancy, Research Funding; Chugai: Honoraria, Research Funding; Daiichi Sankyo: Honoraria, Research Funding; Astra Zeneca: Honoraria, Research Funding; Eisai: Honoraria, Research Funding; Symbio: Research Funding; Ono: Honoraria, Research Funding; Bayer: Honoraria, Research Funding; Solasia: Research Funding; Zenyaku: Research Funding; Incyte: Research Funding; Novartis: Honoraria, Research Funding; Sanofi: Research Funding; HUYA Bioscience: Honoraria, Research Funding; MSD: Honoraria, Research Funding; Astellas Amgen: Honoraria, Research Funding; Abbvie: Honoraria, Research Funding; ARIAD: Research Funding; Takeda: Honoraria, Research Funding; Pfizer: Research Funding; Kyowa Kirin: Honoraria; Nihon Medi-physics: Honoraria; Janssen: Honoraria; Dainihon Sumitomo: Honoraria; Bristol-Byers Squibb: Honoraria; Mundi: Honoraria; Otsuka: Honoraria; Asahi Kasei: Honoraria. Suzumiya:Celgene, Kyowa Kirin, Chugai-Roche, Eisai, Takeda, Celltrion, SymBio, Astellas, Ono, AstraZeneca, Ootsuka, Taiho, Mundi, Dainihon-Sumitomo: Research Funding. Mitsui:MSD pharmaceutical: Research Funding; Maruho pharmaceutical: Research Funding; JCR pharmaceutical: Research Funding; Teijin pharmaceutical: Research Funding; Chugai pharmaceutical: Research Funding; Daiichi Sankyo pharmaceutical: Research Funding; Astellas pharmaceutica: Research Funding; Shionogi pharmaceutical: Research Funding. Kanda:Kyowa Hakko Kirin: Honoraria; Otsuka: Honoraria; Daiichi Sankyo Company: Honoraria; MSD: Honoraria; Chugai: Honoraria; Bristol-Meyers Squib: Honoraria; Novartis: Honoraria; Celgene: Honoraria; Astellas: Honoraria; JCR Pharmaceuticals: Honoraria; Takeda: Honoraria; NextGeM Incorporation: Patents & Royalties: 2019-011392. Atsuta:CHUGAI PHARMACEUTICAL CO., LTD.: Honoraria; Kyowa Kirin Co., Ltd: Honoraria. Suzuki:Celgene: Honoraria; Novartis: Honoraria; AbbVie: Honoraria; Kyowa Hakko Kirin: Honoraria; Chugai Pharmaceutical Co.,Ltd.: Honoraria; Meiji Seika: Honoraria; Bristol-Myers Squibb: Honoraria; Merck Sharp & Dohme: Honoraria; Takeda Pharmaceutical Co., Ltd.: Honoraria; Eisai: Honoraria; ONO Pharmaceutical Co., Ltd.: Honoraria; Janssen: Honoraria.

Congenital macrothrombocytopenia is a genetically heterogeneous group of rare disorders. αIIbβ3 has not been implicated in these conditions. We identified a novel, conserved heterozygous ITGA2B R995W mutation in 4 unrelated families. The surface expression of platelet αIIbβ3 was decreased to 50% to 70% of control. There was spontaneous PAC-1 and fibrinogen binding to resting platelets without CD62p expression. The activation state of αIIbβ3 in 293T cells was higher for αIIb-W995 than for β3-H723 but was weaker than for β3-N562. FAK was spontaneously phosphorylated in αIIb-W995/β3-transfected 293T cells. These results indicate that αIIb-W995/β3 has a constitutive, activated conformation but does not induce platelet activation. αIIb-W995/β3-transfected CHO cells developed membrane ruffling and abnormal cytoplasmic protrusions. The increased size and decreased number of proplatelet tips in αIIb-W995/β3-transduced mouse fetal liver-derived megakaryocytes indicate defective proplatelet formation. We propose that activating mutations in ITGA2B and ITGB3 represent the etiology of a subset of congenital macrothrombocytopenias.

Introduction: Empiric antifungal therapy (EAT) is recommended for persistent or recurrent febrile neutropenia based on an old randomized controlled trial, but such treatment is apparently overtreatment for the majority of patients. On the other hand, preemptive therapy triggered by positive blood tests for fungal antigens and/or imaging study findings was shown to increase the incidence of invasive fungal infection, and thus, a risk-based approach is important. The D-index, which is defined as the area over the neutrophil curve during neutropenia and hence reflects both the duration and depth of neutropenia (Figure 1A), enables real-time monitoring of the risk of invasive fungal infection. Previous studies showed that the cumulative D-index (c-D-index), which was calculated as cumulative D-index from the onset of neutropenia (Figure 1B), had high negative predictive values for invasive mold infection or pulmonary infection with cutoff values of 5,800 or 5,500 in high-risk neutropenic patients [J Clin Oncol 2009; 27: 3849-54. Biol Blood Marrow Transplant 2010; 16: 1355-61]. Methods: We investigated a novel approach, called D-index-guided early antifungal therapy (DET) and compared it to EAT in high-risk neutropenic patients. In the EAT group, empiric antifungal therapy was started for persistent (>=4 days) or recurrent febrile neutropenia. For patients with persistent or recurrent febrile neutropenia in the DET group, preemptive antifungal therapy was applied until c-D-index reached 5,500, but antifungal agent was initiated after c-D-index exceeded 5,500, even if there was no significant finding in serum fungal makers or imaging studies, to prevent excessive invasive fungal infection. Micafungin at 150 mg/day was administered as EAT or DET in this study. We randomized 423 patients who underwent chemotherapy or hematopoietic stem cell transplantation for hematological malignancies, in which predicted period of neutropenia exceeded 7 days, into the EAT group or the DET group, and 413 were eligible for intent-to-treat analyses (201 patients in the EAT group, 212 patients in the DET group). The prophylactic use of fluconazole or itraconazole was allowed. Primary endpoint was the development of proven/probable invasive fungal infection. Results: Backgrounds of the patients were similar between the 2 groups (Table 1). Invasive fungal infection (proven/probable/possible) was observed in 12 patients (6.0%) of the EAT group and 5 patients (2.4%) of DET group, respectively. Proven/probable invasive fungal infection was identified in 5 patients (2.5%) of the EAT group and 1 patient (0.5%) of DET group, which fulfilled the predetermined criteria of non-inferiority of the DET group. Regarding the pathogens, the EAT group included 1 case of candidemia and 4 cases of invasive pulmonary aspergillosis, and the DET group included one fusariosis. The survival rate of the EAT and DET group was 98.0% vs. 98.6% at day 42 and 96.4% vs. 96.2% at day 84, respectively. During the observation period, 31 patients died due to disease progression (n=19), infection (n=5) or other causes (n=7). Causes of infection related mortality included Pseudomonas aerginosa infection (n=2), fusariosis (n=1), toxoplasmosis (n=1) and septic shock by unknown pathogen (n=1). The frequency of micafungin use was significantly lower in the DET group than the EAT group (32.5% vs. 60.2%, P Conclusions: DET successfully reduced the use of antifungal agents without increasing invasive fungal infection or mortality compared to EAT. This randomized controlled study revealed the feasibility of DET in high-risk neutropenic patients. Disclosures Kimura: Astellas: Honoraria; Pfizer: Honoraria; Sumitomo Dainippon Pharma: Honoraria; MSD: Other: Investigator in the institute; Nippon Kayaku: Honoraria; Celgene: Honoraria; Kyowa Hakko Kirin: Honoraria; Takeda: Honoraria. Kanda:Chugai: Consultancy, Honoraria, Research Funding; Shionogi: Consultancy, Honoraria, Research Funding; Nippon-Shinyaku: Research Funding; Ono: Consultancy, Honoraria, Research Funding; MSD: Research Funding; Pfizer: Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria; CSL Behring: Research Funding; Kyowa-Hakko Kirin: Consultancy, Honoraria, Research Funding; Asahi-Kasei: Research Funding; Tanabe-Mitsubishi: Research Funding; Novartis: Research Funding; Astellas: Consultancy, Honoraria, Research Funding; Eisai: Consultancy, Honoraria, Research Funding; Otsuka: Research Funding; Dainippon-Sumitomo: Consultancy, Honoraria, Research Funding; Sanofi: Research Funding; Taisho-Toyama: Research Funding; Taiho: Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria; Mochida: Consultancy, Honoraria; Alexion: Consultancy, Honoraria; Takara-bio: Consultancy, Honoraria. Fujiwara:Shire: Consultancy; Pfizer: Consultancy; Chugai: Consultancy; Kirin: Consultancy; Kyowa-Hakko: Consultancy; Astellas: Consultancy. Suzumiya:Celltrion: Research Funding; Taiho: Research Funding, Speakers Bureau; SymBio: Research Funding; Toyama Chemical: Research Funding; Takeda: Research Funding, Speakers Bureau; Eisai: Research Funding, Speakers Bureau; Chugai-Roche: Research Funding, Speakers Bureau; Kyowa Hakko Kirin: Research Funding, Speakers Bureau; Zenyaku Kogyo: Consultancy; Abbvie: Consultancy, Speakers Bureau; Janssen: Consultancy, Speakers Bureau; Celgene: Consultancy, Speakers Bureau; Pfizer: Research Funding; Sumitomo Dainioppon: Research Funding, Speakers Bureau; Astellas: Research Funding, Speakers Bureau; Bristol Myers Squibb: Speakers Bureau; Nippon Shinyaku: Speakers Bureau; Ono: Speakers Bureau; Ohtsuka: Speakers Bureau; Shire Japan: Speakers Bureau. Takamatsu:Taisho Toyama Pharmaceutical: Research Funding; TAIHO Pharmaceutical: Research Funding; Pfizer: Research Funding; Bristol-Myers Squibb: Research Funding; Ono Pharmaceutical: Research Funding; Astellas Pharma: Research Funding; Kyowa Hakko Kirin: Research Funding; Chugai Pharma: Research Funding; Takeda Pharmaceutical: Research Funding; Celgene: Honoraria. Tamura:Astellas Phrma: Research Funding; Eisai: Speakers Bureau; Kyowa Hakko Kirin: Speakers Bureau; Ono Pharmaceutical: Speakers Bureau.

Background: The immunological graft-versus-leukemia effects associated with chronic myeloid leukemia (CML) are well-established. Both regulatory T cells (Treg) and natural killer (NK) cells play a role in the maintenance of CML. In this study, we examined changes in the number of circulating Treg and NK cells and the NK activity in CML patients before and after treatment with a tyrosine kinase inhibitor (TKI). Our goal was to evaluate the clinical significance of this treatment on these two different cell types. Moreover, the levels of these cells suggest that there is a need to make the TKI more effective as an immunotherapy. Patients and Methods: Treg and NK cells were characterized and quantified by flow cytometry in 27 newly diagnosed CML patients, 19 newly diagnosed patients with other myeloproliferative neoplasms (MPN), and 15 healthy controls (HC-group). CML patients, who were diagnosed as chronic phase and initially treated with a TKI between September 2007 and July 2017 and were followed up for more than 12 months were enrolled in the study. The molecular response was analyzed by quantitative PCR of the BCR-ABL fusion genes at approximately 6-month intervals after the initiation of treatment. After obtaining informed consent, peripheral blood (PB) was drawn from all patients at their initial diagnosis and at 6-month intervals thereafter. CD4+CD25+Foxp3+ Treg cells, CD3+CD4+T cells, CD3+CD8+ T cells, and CD3-CD56+ NK cells were analyzed using flow cytometry, and the absolute numbers of these cells were calculated. The NK activity was measured by the standard 51Cr release assay at an effector:target (E:T) ratio of 20:1. Results: The median age for CML patients was 65 years (range: 44-85), and 17 out of 27 of the patients were male. The median period of observation was 656 days. After 12 months, 12 CML patients (MMR-group) had a major molecular response (MMR), and 15 patients (NR-group) did not have a MMR. Treg% values in the PB before treatment in the CML-group (0.39±0.30%) were significantly lower than that in the HC-group (0.70±0.29%, p Conclusions: The absolute number of Treg cells in the CML patients decreased remarkably after treatment. The absolute number and the percentage of Treg cells decreased remarkably after treatment especially among the CML patients in the MMR-group. Although the number of NK cells did not change in the MMR-group compared to that in the HC-group, the NK activity was significantly lower and tended to approach the value of the normal control group after treatment. These data suggest that patients with CML have a more profound remission by treatment with a TKI resulting in decreased levels of Treg cells and increased NK activity. Immunological control via T cells and NK cells is very important in the treatment of CML. Disclosures Suzumiya: Chugai-Roche: Research Funding, Speakers Bureau; Kyowa Hakko Kirin: Research Funding, Speakers Bureau; Pfizer: Research Funding; SymBio: Research Funding; Ono: Speakers Bureau; Nippon Shinyaku: Speakers Bureau; Eisai: Research Funding, Speakers Bureau; Shire Japan: Speakers Bureau; Taiho: Research Funding, Speakers Bureau; Toyama Chemical: Research Funding; Celltrion: Research Funding; Takeda: Research Funding, Speakers Bureau; Zenyaku Kogyo: Consultancy; Abbvie: Consultancy, Speakers Bureau; Janssen: Consultancy, Speakers Bureau; Celgene: Consultancy, Speakers Bureau; Sumitomo Dainioppon: Research Funding, Speakers Bureau; Astellas: Research Funding, Speakers Bureau; Bristol Myers Squibb: Speakers Bureau; Ohtsuka: Speakers Bureau.

Background: CT-P10 is the first biosimilar to innovator rituximab (RTX) approved for all indications by the European Medicines Agency. The pharmacokinetics (PK) equivalence and non-inferior efficacy of CT-P10 compared with RTX in patients with newly diagnosed advanced follicular lymphoma (FL) was previously demonstrated when used with combination chemotherapy of cyclophosphamide, vincristine and prednisone (CVP) (Kim WS et al,. 2017). Objectives: This is an ongoing, phase 3, randomized, double blind, active controlled study to demonstrate similarity of efficacy and safety between CT-P10 and RTX as monotherapy in patients with newly diagnosed FL with low tumor burden (NCT02260804). The results of efficacy as determined by overall response rate (ORR), PK, pharmacodynamic, safety and immunogenicity up to 7 months (prior to Maintenance Cycle 3) are presented here. Methods: Patients with FL with stage II-IV, Grade 1-3a and low tumor burden as assessed by GELF criteria were randomized in a 1:1 ratio to receive either CT-P10 or RTX (375mg/m2 i.v.) monotherapy weekly for 4 weeks as induction therapy, followed by maintenance therapy every 2 months for 2 years. Patients who had complete response (CR), unconfirmed CR, partial response or stable disease after the induction therapy were eligible for maintenance therapy. The primary endpoint was ORR over 7 months according to the IWG criteria and was assessed by the independent review committee as central review and by the site investigator as local review. Results: In total, 258 patients were randomized to either the CT-P10 (n=130) or RTX (n=128) group with well-balanced baseline characteristics. Overall, 231 patients (n=119/130 [92%] and n=112/128 [88%] in the CT-P10 and rituximab groups, respectively) completed up to 7 months. Therapeutic similarity between CT-P10 and RTX was established by meeting the predefined margin of ±17% for the primary efficacy endpoint of ORR in the intent-to-treat (ITT) and per-protocol (PP) population (Figure 1). An ORR of 83.1% and 81.3% for CT-P10 and RTX, respectively, was observed over 7 months. The ORR difference was 1.8% and 90% confidence interval (CI) of the treatment difference estimate was entirely within the equivalence margin; 90% CI (-6.43, 10.20). Similar PK profiles were shown in the CT-P10 and RTX. Maximum and trough concentration of CT-P10 or RTX at each cycle were similar between the two groups. Median number of B-cells decreased to the lower limit of quantification ( CT-P10 was well tolerated and the safety profile of CT-P10 over 7 months was consistent with that of RTX, with no new, unexpected safety findings. Treatment-emergent adverse events (TEAEs) were reported in 71% and 67% patients in the CT-P10 and RTX, respectively. Adverse events of special interest included infections and infusion-related reactions occurred in a similar proportion of patients in each treatment group (Table 1). Neither progressive multifocal leukoencephalopathy nor Hepatitis B virus reactivation was reported in either group. The proportion of patients with positive anti-drug antibody were similar between 2 groups (0.8% vs. 2.3%) over 7 months. Among them, neutralizing antibody were detected in one patient in the CT-P10 group. Conclusions: This study demonstrates therapeutic equivalence of CT-P10 to RTX in previously untreated LTBFL. CT-P10 was well-tolerated and the safety profile including immunogenicity of CT P10 was comparable to that of RTX over 7 months. Disclosures Ogura: SymBio: Research Funding; Mundi Pharma: Consultancy; Celltrion: Consultancy, Research Funding; Takeda: Honoraria; Cellgene: Honoraria; MeijiSeika Pharma: Consultancy. Sancho:SERVIER: Honoraria; SANOFI: Honoraria; CELGENE: Honoraria; KERN FHARMA: Honoraria, Speakers Bureau; GILEAD: Honoraria, Research Funding; JANSSEN: Honoraria, Speakers Bureau; ROCHE: Honoraria, Speakers Bureau; MUNDIPHARMA: Honoraria. Lennard:Janssen: Membership on an entity's Board of Directors or advisory committees; Roche: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Research Funding. Jurczak:European Medicines Agency: Consultancy; Astra Zeneca/Acerta: Consultancy, Research Funding; Sandoz-Novartis: Consultancy; Janssen: Consultancy, Research Funding; Gilead: Consultancy, Research Funding; Afimed: Research Funding; Bayer: Research Funding; Beigene: Research Funding; Celgene: Research Funding; Epizyme: Research Funding; Nordic Nanovector: Research Funding; Merck: Research Funding; Morphosys: Research Funding; Pharmacyclics: Research Funding; Servier: Research Funding; Roche: Research Funding; TG Therapeutics: Research Funding. Coiffier:CELGENE: Consultancy, Membership on an entity's Board of Directors or advisory committees; MUNDIPHARMA: Membership on an entity's Board of Directors or advisory committees; CELLTRION: Membership on an entity's Board of Directors or advisory committees; MORPHOSYS: Membership on an entity's Board of Directors or advisory committees; NOVARTIS: Membership on an entity's Board of Directors or advisory committees. Buske:Roche: Honoraria, Research Funding; Bayer: Research Funding; Janssen: Honoraria, Research Funding. Lee:Celltrion, Inc: Employment.

Background: It is important to determine the changes in both donor T cells and B cells on immune reconstitution following allogeneic hematopoietic stem cell transplantation (HCT). It is well-known that Treg cells play important roles in the prevention of T-cell-mediated graft-versus-host disease (GVHD) and in promoting tumor escape from T-cell-dependent immunosurveillance. However, very little is known about the number and function of both CD4+CD25+Foxp3+ regulatory T (Treg) cells and CD3-CD56+ natural killer (NK) cells and about NK activity during immune reconstitution. In this study, we examined changes in the number of circulating Treg and NK cells in the peripheral blood (PB) and the NK activity of HCT patients at various times after HCT, and we evaluated the clinical significance of these findings relative to patient survival. Patients and Methods: We evaluated 29 consecutive patients (ages >18) without GVHD who underwent HCT for different hematologic diseases between December 2008 and April 2017. Treg and NK cells were characterized and quantified by flow cytometry from these 29 patients and 15 healthy controls. Evaluated variables included recipient and donor characteristics, transplant-related factors, including conditioning regimen (myeloablative conditioning or reduced-intensity conditioning), donor type (matched sibling donor, unrelated donor, other relative (haploidentical), or cord blood transplant), degree of match (8/8, 7/8, 6/8, or haploidentical), and GVHD prophylaxis. Patients who were followed up for more than 12 months were enrolled in the study. After obtaining informed consent, blood was drawn from all patients on the day of engraftment and at day 100 and at 12-month intervals thereafter. CD4+CD25+Foxp3+ Treg cells, CD3+CD4+T cells, CD3+CD8+ T cells, and CD3-CD56+ NK cells were analyzed using flow cytometry, and the absolute numbers of these cells were calculated. The NK activity was measured by the standard 51Cr release assay at an effector: target (E: T) ratio of 20:1. Results: The median patient age was 58 years (range: 19-71 years), and 21 out of 29 of the patients were male. At 100 days, the percentage of B cells (2.5±6.0%) and absolute numbers of CD8+ T cells (269.7±284.8/μL), CD4+T (22.83±292.4/μL) cells and NK cells (248.3±229.1/μL) were significantly lower than those at 2 years (20.9±11.6%, 747.2±648.4/μL, 588.0±607.3/μL, 558.1±336.2/μL, p Conclusion: The percentage and the absolute number of Treg cells in HCT patients were significantly lower than those in the normal control group, whereas CD8+ T cells was significantly higher in the patients within 2 years after HCT than that in the normal control group. The % Treg cells did not recover even at 3 years after HCT, rather it tended to be somewhat lower. In contrast, there was no change in the NK cells. These results suggest that the immunological reconstitution has not been achieved even at 3 years after transplantation and that the immunological consequences of GVHD are maintained. The antitumor effect is also maintained. On the other hand, NK cells recover at an early stage. The roles of these different immune cells after HCT require further investigation. Disclosures Suzumiya: Eisai: Research Funding, Speakers Bureau; Pfizer: Research Funding; Celltrion: Research Funding; Taiho: Research Funding, Speakers Bureau; Sumitomo Dainioppon: Research Funding, Speakers Bureau; Celgene: Consultancy, Speakers Bureau; SymBio: Research Funding; Bristol Myers Squibb: Speakers Bureau; Toyama Chemical: Research Funding; Chugai-Roche: Research Funding, Speakers Bureau; Kyowa Hakko Kirin: Research Funding, Speakers Bureau; Zenyaku Kogyo: Consultancy; Abbvie: Consultancy, Speakers Bureau; Janssen: Consultancy, Speakers Bureau; Astellas: Research Funding, Speakers Bureau; Takeda: Research Funding, Speakers Bureau; Nippon Shinyaku: Speakers Bureau; Ono: Speakers Bureau; Ohtsuka: Speakers Bureau; Shire Japan: Speakers Bureau.

Adult T-cell leukemia/lymphoma (ATL) is a malignancy of mature T lymphocytes caused by human T-lymphotropic virus type I. Intensive combination chemotherapy and allogeneic hematopoietic stem cell transplantation have been introduced since the previous Japanese nationwide survey was performed in the late 1980s. In this study, we delineated the current features and management of ATL in Japan. The clinical data were collected retrospectively from the medical records of patients diagnosed with ATL between 2000 and 2009, and a total of 1665 patients' records were submitted to the central office from 84 institutions in Japan. Seventy-one patients were excluded; 895, 355, 187, and 157 patients with acute, lymphoma, chronic, and smoldering types, respectively, remained. The median survival times were 8.3, 10.6, 31.5, and 55.0 months, and 4-year overall survival (OS) rates were 11%, 16%, 36%, and 52%, respectively, for acute, lymphoma, chronic, and smoldering types. The number of patients with allogeneic hematopoietic stem cell transplantation was 227, and their median survival time and OS at 4 years after allogeneic hematopoietic stem cell transplantation was 5.9 months and 26%, respectively. This study revealed that the prognoses of the patients with acute and lymphoma types were still unsatisfactory, despite the recent progress in treatment modalities, but an improvement of 4-year OS was observed in comparison with the previous survey. Of note, one-quarter of patients who could undergo transplantation experienced long survival. It is also noted that the prognosis of the smoldering type was worse than expected.

A new World Health Organization classification was recently proposed. However, classification of peripheral T-cell lymphomas remains to be clarified. Particularly, unspecified type was considered as a heterogeneous category. Here we studied the expressions of chemokine receptors, Th1-associated CXCR3 and CCR5 and Th2-associated marker ST2(L), and activated T-cell receptor OX40/CD134 in 185 patients with nodal T-cell lymphoma, and evaluated the relationship to prognosis. Their expression patterns correlated with the specific subtype of nodal T-cell lymphoma, such as angioimmunoblastic T-cell lymphoma (AILD), anaplastic large cell lymphoma (ALCL), and in peripheral T-cell lymphoma (PTCL), unspecified. In AILD, almost all cases were immunoreactive for OX40/CD134 (96%) and for CXCR3 (89%). In ALCL, all cases were immunonegative for OX40/CD134, and only a few cases (24%) were immunoreactive for CXCR3, whereas almost all cases (94%) were positive for ST2(L). Cases of PTCL, unspecified, were divided into 2 groups; group 1 (cases positive for either ST2(L), CCR5, or CXCR3) tended to show favorable prognosis compared with group 2 (cases negative for ST2(L), CCR5, and CXCR3). Our results indicate that further subtyping of PTCL, unspecified, into groups 1 and 2 could be significant for evaluating prognosis and understanding the functional role of these tumors.

De novo CD5+ diffuse large B-cell lymphoma (CD5+ DLBCL) is known to have phenotypically and genotypically different characteristics than CD5- DLBCL and mantle cell lymphoma (MCL). To further characterize CD5+ DLBCL, 109 patients with CD5+ DLBCL were reviewed, and the results were compared with those of 384 CD5- DLBCL and 128 cyclin D1+ MCL patients. Patients with CD5+ DLBCL showed a higher age distribution (median, 66 years; P =.0083) and a female predominance (male-female ratio, 49:60, P =.011) compared with those with CD5- DLBCL. CD5+ DLBCL was more closely associated with many aggressive clinical features or parameters than CD5- DLBCL: 69% older than 60 years (P =.039), 34% with performance status greater than 1 (P =.0016), 69% with serum lactate dehydrogenase level higher than normal (P

Background: Adult T-cell leukemia-lymphoma (ATL), which is a hematological malignancy related to human T-lymphotropic virus type I (HTLV-1), is known as a malignant lymphoid disease with poor prognosis. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is considered a treatment modality to contribute prolonging the survival in some population of patients (Katsuya H et al. 2015 Blood). A small study indicated that ATL cells frequently have chromosomal abnormalities including various numerical aberrations and complex structural abnormalities (Kamada N et al. Cancer Res. 1992). However, there has been no large study to examine the relationship between chromosomal abnormalities and survival. Here we report the impact of chromosomal abnormalities on survival in ATL patients with conventional chemotherapy. Patients and Methods: In this study, we extracted a population of patients from the database of ATL-PI project, which is a nation-wide survey of ATL patients newly diagnosed between January 2000 and May 2009, and the overall results were reported elsewhere (Katsuya H et al. JCO 2012, Blood 2015). Fifteen hundred and twelve patients were registered in this project, and we excluded the patients who received allo-HSCT, and who were diagnosed with smoldering and chronic type ATL. Moreover, patients whose chromosomal analysis with G-banding stain showed normal, multiploid or no mitosis, or no information on chromosomal analysis and on the factors to determine the ATL-PI were also excluded. As a consequence, 210 patients with acute (n=157) and lymphoma (n=53) types were selected for this analysis. Abnormal karyotypes were analyzed from the aspect of numerical and structural break points. To identify the specific abnormalities and to avoid the secondary changes, the stem-line karyotype, which meant the simplest abnormal clone, was used as the representative karyotype for this study. A statistical analysis was performed with 'EZR' (Kanda T. BMT 2013). We examined an overall survival (OS) in relation to chromosomal abnormalities using cox proportional hazard regression model, and p-value of less than 0.05 was considered as statistically significant. Results: Among the 210 patients, 119 and 91 patients were male and female, respectively. The median age of patients was 66 years (40-89). With respect to other factors of ATL-PI, the median level of soluble interleukin-2 receptor (sIL-2R), serum albumin, were 30800 U/mL (620-1,219,000), 3.6 g/dL (1.6-5.3), respectively. Almost all patients were diagnosed with stage II-IV of clinical stage (n=200), and almost half of patients certified 2-4 of ECOG-PS (n=113). Sixty-nine, 106, and 35 patients were classified as high, intermediate and low risk by ATL-PI, respectively. Specimens used for chromosomal analysis were taken from lymph nodes (n=77), bone marrow (n=72), peripheral blood (n=50), pleural fluids (n=4), ascites (n=4) and 1 each of stomach, sinusoidal tumor, and tonsillar lesions. The median number of numerical abnormalities and marker chromosomes were 2 (0-20) and 1 (0-20), and the median number of structural break points was 6 (0-21). The numerical abnormalities were loss of sex chromosomes (31%) followed in decreasing order by -14 (24%), -13 (19%), -9 (12%) and -17 (12%) while the structural break points were located at 1p (30%), 3q (30%), 14q (30%), 6q (26%) and 1q (22%). The median OS was 286 days (1-2565). ECOG-PS, serum albumin level, sIL-2R, and structural break points at 9q or 19p, and the number of structural break points over 4 or 5 points were shown to be the significant factors affecting the survival by univariate analysis. Then, the multivariate analysis indicated that 2-4 of ECOG-PS (HR: 2.021, 95%CI: 1.436-2.791, p Discussions and Conclusion: This is the first large study showing the impact of chromosomal abnormality in acute and lymphoma types ATL on OS. It demonstrated that structural break points at 9q and 19p were the independent risk factors for OS in addition to those of ATL-PI. Disclosures Katsuya: The Uehara Memorial Foundation: Research Funding. Utsunomiya:Daiichi Sankyo Co., Ltd.: Speakers Bureau. Tsukasaki:Daiichi Sankyo Co., Ltd.: Consultancy; Takeda: Research Funding. Suzumiya:Takeda: Honoraria; Astellas: Research Funding; Kyowa Hakko kirin: Research Funding; Chugai: Honoraria, Research Funding; Toyama Chemical: Research Funding; Eisai: Honoraria, Research Funding.

[Background] Mantle cell lymphoma (MCL) is an aggressive B-cell lymphoma, characterized by the overexpression of cyclin D1 derived from t(11;14)(q13;q32) and poor prognosis. Most MCLs show nodal presentation, but also accompany extranodal involvement, such as bone marrow, peripheral blood or gastrointestinal tract. As a result, many MCLs present with advanced stage disease. Since only a small portion of patients show limited-stage disease, minimal data exist on treatment of patients diagnosed with limited stage disease. Nevertheless, the treatment strategy of MCL is recommended according to the clinical stage of limited- (stage I or non-bulky II) vs. advanced-stage, as well as other types of lymphoma. [Patients and methods] We recently collected 633 patient data of MCL (Chihara, et al. Ann Oncol 2015). Information of clinical stage was available in 626 patients. The patient data were retrospectively analyzed the by the clinical stage at initial presentation. [Results] The clinical stage was I in 24 patients (4%), II in 33 (5%), III in 70 (11%), and IV in 499 (80%). Only one patient presented with bulky stage II. Detailed demographic information by the clinical stage are listed in Table. Age and sex were not significantly different by clinical stage. Limited stage patients were associated with better performance status (PS), less B symptoms, no extranodal involvement, and lower lactate dehydrogenase (LDH) level and white blood cell (WBC) count. Most patients in any stage were treated with cytotoxic chemotherapy, but more patients in limited stage received radiotherapy. The proportion of high-dose cytarabine (HDCA)-containing regimen over CHOP/CHOP-like was higher in advanced stage patients. Complete and overall response rates were 92% and 96% in stage I, 58% and 94% in stage II, 66% and 86% in stage III, and 52% and 82% in stage IV, respectively (P = 0.02). However, the higher response rate in limited stage patients did not translate into better prognosis. The median survival was 11.0 years in stage I, 13.4 years in stage II, 11.5 years in stage III, and 5.6 years in stage IV (Figure). The prognosis was not significantly different among patients with stage I, II, and III (P = 0.33). [Conclusion] Prognosis of limited-stage MCL was almost similar to that of stage III MCL. Although the present study includes several limitations including a retrospective nature and limited number of patients, prognosis of patients with limited-stage MCL was not satisfactory. The significance of radiotherapy, as well as the optimal choice of chemotherapy, for limited-stage MCL needs re-evaluation. Table Table. Figure Figure. Disclosures Suzuki: Chugai: Honoraria; Kyowa Hakko kirin: Honoraria; Bristol-Myers Squibb: Honoraria. Asano:Jannsen: Honoraria; Chugai: Honoraria. Kinoshita:Ono: Research Funding; Gilead: Research Funding; Zenyaku: Honoraria, Research Funding; Takeda: Research Funding; Chugai: Honoraria, Research Funding; Eisai: Honoraria, Research Funding; Solasia: Research Funding; Janssen: Honoraria; Kyowa Kirin: Honoraria. Suzumiya:Chugai: Honoraria, Research Funding; Astellas: Research Funding; Eisai: Honoraria, Research Funding; Takeda: Honoraria; Toyama Chemical: Research Funding; Kyowa Hakko kirin: Research Funding. Ogura:SymBio Pharmaceuticals: Consultancy, Honoraria; Celltrion, Inc.: Consultancy, Honoraria.

Background: Chronic lymphocytic leukemia (CLL) is the most frequent leukemia in the Western countries but very rare in the East Asia including Japan. The previous reports from Japan showed that CLL patients in Japan have different characteristics including atypical morphology and immunophenotypes compared to those of Western countries. We started the cooperative study of CLL between Japan and Austria from 2012. Aim: The aim of this retrospective study is to compare the characteristics of typical CLL patients in Japan and Austria. Material and Methods: Diagnostic procedures were first harmonized between sites and laboratories. Samples were exchanged between Japan and Austria, and morphology, flow cytometry (FCM), FISH and IGVH gene sequence were independently assessed. After technical agreement, we selected 86 Austrian CLL patients from the database of the Medical University of Vienna, and 80 Japanese patients from the CLLRSG-01 prospective observational study. All CLL cases had typical immunophenotypes (Matutes score 4 or 5). The analyses of morphology, FCM and FISH including the deletion in 11q22, 13q14, and 17p13, and trisomy 12 were performed. Immunoglobulin heavy chain variable region (IGHV) gene mutation was also analysed. Results: Median age of CLL patients was 62.5 years old (range: 35-93) in Austria and 66.4 years (range: 34-93) in Japan. Japanese patients were significantly older at diagnosis than Austrian patients (P = 0.027). No differences were found for sex and the Binet stage. Also, there were no differences in laboratory findings (lymphocyte and platelet counts and serum LDH and beta-2-microglobulin levels) except for hemoglobin (Hb) level (median: 13.8g/dL in Austria vs. 13.1g/dL in Japan, P = 0.005) between two countries. Immunophenotypic study by FCM showed higher proportion of CD38 expression in Austrian CLL patients (34% in Austria vs. 10% in Japan, P < 0.001). No differences were found for chromosomal aberrations by FISH analyses between Austria and Japan. del(13q14) was found in 62% and 56% (P = 0.456), trisomy 12 in 7% and 13% (P = 0.315), del(11q22) in 13% and 10% (P = 0.513) and del(17p13) in 7% and 3% (P = 0.179), in Austria and Japan, respectively. The IGHV mutation rate was higher in Japan (P = 0.002). Mutated IGHV was found in 61% of Austrian and 84% of Japanese patients. IGHV family usage profile was significantly different. In Austrian patients, VH1, 2, 3, 4, 5, 6, and 7 were 24%, 1%, 54%, 19%, 1%, 1% and 0%, respectively. However, in Japanese patients, they were 2%, 0%, 75%, 20%, 2%, 0% and 2%, respectively. The proportion of VH1 was surprisingly lower in Japan (P = 0.006). Notably, none of the Japanese patients used VH1-69, in contrast to the use of 10% in Austrian patients. Conclusion : Our analysis indicates that there are clear differences between Japanese and European CLL patients, in age, Hb level, CD38 expression and IGHV usage and mutations. The reason for particular IGHV usage should further be investigated. Disclosures Takizawa: Teijin: Research Funding; Takeda: Honoraria, Research Funding; Celgene: Honoraria; Chugai: Honoraria, Research Funding; Kyowa Hakko Kirin: Honoraria, Research Funding; Janssen: Honoraria; Sumitomo Dainippon: Honoraria, Research Funding; MSD: Honoraria, Research Funding. Suzuki:Bristol-Myers Squibb: Honoraria; Kyowa Hakko kirin: Honoraria; Chugai: Honoraria. Hoermann:Gilead: Research Funding; Novartis: Honoraria; Amgen: Honoraria; Ariad: Honoraria. Aoki:SymBio Pharmaceuticals: Consultancy. Suzumiya:Kyowa Hakko kirin: Research Funding; Eisai: Honoraria, Research Funding; Astellas: Research Funding; Takeda: Honoraria; Toyama Chemical: Research Funding; Chugai: Honoraria, Research Funding. Jaeger:Janssen: Consultancy, Honoraria, Other: Travel, Accommodations, Expenses.

Background and Objectives: Approximately 10% of cases of diffuse large B cell lymphoma have a MYC rearrangement (MYC+ DLBCL) and these have been associated with an inferior outcome following standard therapy. Double-hit (DHL) and triple-hit (THL) lymphomas harbor concurrent rearrangements of MYC and BCL2 and/or BCL6 are associated with a very poor outcome. The role of hematopoietic stem cell transplantation (HSCT) for MYC+ DLBCL remains unclear. Autologous (Auto-) HSCT could not salvage chemosensitive relapse of MYC+ DLBCL in the CORAL study. However, other two retrospective studies showed the efficacy of Auto-HSCT as consolidation therapy for DHL who attained first complete response (CR), in contrast to the poor outcome of patients without HSCT. A retrospective study of HSCT for MYC+ DLBCL using Japanese registry-based data was conducted. Materials and Methods: The data of Japanese HSCT registry were surveyed, and 54 patients with MYC+ DLBCL were identified from 4,336 adult patients with DLBCL. These pathological and cytogenetic diagnoses including G-banding and/or fluorescence in situ hybridization (FISH) were carried out at each institution. The questionnaires were sent to collect patient data of clinical presentation and therapies, as well as pathology and cytogenetic reports. This study was approved by the ethical committees of the Japan Society for Hematopoietic Cell Transplantation (JSHCT) and Shimane University. Results: A total of 48 patient data were obtained from 36 institutions. MYC rearrangements were identified by G-banding in 44 patients and FISH in 4 patients. The median age at diagnosis was 55 (range, 21-67) years. Twenty patients were male. Six patients were transformed DLBCL from indolent lymphoma. Eleven patients had CNS involvement during clinical course. Twenty seven patients were single hit lymphoma (SHL, only MYC), 20 patients were DHL (MYC+BCL2; n = 15, MYC+BCL6; n = 5) and one patient was THL (MYC+BCL2+BCL6). Thirty-one patients (64.6%) received R-CHOP as initial therapy. The CR rate after the initial therapy was 66.7% in SHL, but was 28.6% in DHL/THL. In total, data were available for 32 Auto-HSCTs and 17 allogeneic (Allo-) HSCTs including one after relapsed of Auto-HSCT were available between 2003 and 2012 and subjected for further analysis. Median follow up time from HSCT in surviving patients was 17.1 months (range, 9.8-130.0). In Auto-HSCT in first CR, median time from diagnosis to HSCT was 6.5 months (range, 4.6-20.9). The overall survival (OS) at 2 years, the event free survival (EFS) at 2 years, the relapse rate at 1 year and the non-relapse mortality (NRM) at 1 year was 71.1% (95% CI: 51.7-83.9), 65.5% (95% CI: 46.4-79.2), 31.4% (95% CI: 16.2-47.9) and 3.1% (95% CI: 0.2-14.0), respectively. The outcome of SHL (n = 20) and DHL (n = 12) were almost similar (OS at 2 years; 73.8% vs 66.7%, P = 0.62). The disease status at HSCT was a significant adverse prognostic factor. OS of patients who received HSCT in CR (n = 23) was 82.4%, and was significantly better than that in partial response (PR, 42.9%, P = 0.04). The outcome of HSCT in first CR (n = 14) and second CR (n = 9) was similar (respective OS at 2 years; 78.6% and 88.9%, P = 0.57). In Allo-HSCT, four patients (23.5%) had poor performance status (2-4), eleven patients (64.7%) were chemoresistant prior to HSCT, and 13 patients (76.5%) received reduced intensity conditioning regimen. The median time from diagnosis to HSCT was 10.0 months (range, 2.3-54.5). Their OS at 2 years, EFS at 2 years, relapse rate at 1 year and NRM at 100 days were 29.4% (95% CI: 10.7-51.1), 17.6% (95% CI: 4.3-38.3), 41.2% (95% CI: 17.6-63.5) and 41.2% (95% CI: 17.1-64.0), respectively. The outcome of SHL (n = 8) and DHL/THL (n = 9) was almost similar (OS at 2 years; 37.5% vs 22.2%, P = 0.78). Conclusions: Auto-HSCT as consolidation therapy for MYC+ DLBCL was effective, as reported elsewhere. Auto-HSCT for chemosensitive relapse was also effective, in contrast to the CORAL study. The efficacy of Allo-HSCT was limited, particularly for chemoresistant patients. Although the present study include several limitations of retrospective nature, HSCT is not recommended for chemoresistant MYC+ DLBCL. Disclosures Suzuki: Bristol-Myers Squibb: Honoraria; Kyowa Hakko kirin: Honoraria; Chugai: Honoraria. Izutsu:Abbvie: Research Funding; Gilead: Research Funding; Celgene: Research Funding; Janssen Pharmaceutical K.K.: Honoraria; Eisai: Honoraria; Kyowa Hakko Kirin: Honoraria; Chugai Pharmaceutical: Honoraria, Research Funding; Takeda Pharmaceutical: Honoraria; Mundipharma KK: Research Funding. Kuroda:Bristol Myers Squibb: Honoraria, Research Funding; Astra Zeneca: Research Funding; Celgene: Honoraria, Research Funding; Janssen: Honoraria. Suzumiya:Astellas: Research Funding; Takeda: Honoraria; Chugai: Honoraria, Research Funding; Kyowa Hakko kirin: Research Funding; Toyama Chemical: Research Funding; Eisai: Honoraria, Research Funding.

Combinations of bortezomib and novel targeted therapeutic agents may synergistically increase antitumor effect and may overcome specific cellular resistance and/or antiapoptotic processes. However, highly effective combinations of bortezomib with targeted therapeutic agents have not been developed to date. We examined the synergistic effects of various compounds and bortezomib on myeloma cells and identified fusicoccin derivative ISIR-042 as the most potent drug. We developed ISIR-042 that exerts higher cytotoxic effects on hypoxic cells than on normoxic cells and that preferentially inhibits stem/progenitor cells in pancreatic cancer cell lines compared with other chemotherapeutic agents (Kawakami et al, Anticancer Agents Med Chem 2012). In the present study, we determined the synergistic effects of bortezomib and ISIR-042 cotreatment on human multiple myeloma cells. Human multiple myeloma cell lines RPMI8226, KMS-11, and KMS-26 were cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% CO2 in air. Viability of cells cultured with or without the test compounds for indicated time was examined by performing a modified MTT assay.Bone marrow specimens and ascitic fluids were collected from patients at diagnosis or relapse after obtaining written informed consent for sample collection in accordance with institutional policy. The study was approved by the Institutional Review Board of Shimane University. For performing colony-forming assay, the cells (1 × 104/dish) were plated in 1.1 ml semisolid methylcellulose medium supplemented with 0.8% methylcellulose and 20% FBS in triplicates for 10 days. CHOP-/- and CHOP+/+ MEF cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% FBS. Expression of cell surface antigens was analyzed by performing flow cytometry with anti-CD38 or anti-CD138 antibody. Combined effects of myeloma cells in primary culture were analyzed by performing flow cytometry with anti-CD19 and anti-CD138 antibodies. Expression of caspase-10, BCL-2, p62, BCLAF1, LC3, actin, and CHOP was determined by performing western blotting with respective monoclonal antibodies. mRNA expression of genes encoding XBP-1u and XBP-1s was analyzed by performing reverse transcription-PCR. CHOP mRNA levels were measured by performing real-time PCR. ISIR-042 inhibited the growth of RPMI8226 cells in a concentration-dependent manner and exerted synergistic effects with bortezomib. ISIR-042 also exerted synergistic effects with carfilzomib. Results of flow cytometry showed that the combination of ISIR-042 and bortezomib decreased the number of myeloma cells in the primary culture in a concentration-dependently. Colony-forming assay showed that RPMI8226 cells were more sensitive than normal mouse hematopoietic cells to ISIR-042 and bortezomib cotreatment. ISIR-042 decreased the number of CD38-CD138- cells and increased the number of CD38+CD138+ cells in KMS11 cell population. mRNA expression of genes encoding XBP-1u and XBP-1s was not affected in RPMI8226 cells treated with ISIR-042 or bortezomib alone. However, mRNA expression of the gene encoding XBP-1s significantly increased in RPMI8226 cells cotreated with ISIR-042 and bortezomib. Growth-inhibitory effects of ISIR-042 and bortezomib were not associated with caspase-10 which modulates autophagic response for survival. Expression of autophagic markers such as LC3 and p62 was also examined. ISIR-042 exerted modest effects in the presence or absence of bortezomib. ER stress-mediated CHOP induction promotes the cytotoxic effects of proteasome inhibitors on many cancer cells. Carfilzomib and ISIR-042 cotreatment significantly induced CHOP mRNA expression, suggesting that ISIR-042 and proteasome inhibitor cotreatment induced apoptosis by enhancing ER stress and activating CHOP. This was confirmed using CHOP-/- MEF cells. Our results showed that carfilzomib and ISIR-042 cotreatment did not exert cytotoxic effects on CHOP-/- MEF cells. These results suggest that treatment of multiple myelomas with ISIR-042-supplemented proteasome inhibitor-based chemotherapy exerts beneficial anticancer effects. Disclosures Suzuki: Chugai: Honoraria; Bristol-Myers Squibb: Honoraria; Kyowa Hakko kirin: Honoraria. Suzumiya:Astellas: Research Funding; Takeda: Honoraria; Eisai: Honoraria, Research Funding; Kyowa Hakko kirin: Research Funding; Chugai: Honoraria, Research Funding; Toyama Chemical: Research Funding.

Therapy-related acute myeloid leukemia and myelodysplastic syndrome (t-AML/MDS) represent severe late effects in patients receiving hematopoietic cell transplantation (HCT) for malignant lymphoma (ML). The option of high-dose therapy with autologous HCT or allogeneic HCT with reduced-intensity conditioning remains controversial in patients with relapsed ML. We retrospectively analyzed incidence, outcome, and risk factors for the development of t-AML/MDS in ML patients treated with autologous or allogeneic HCT. A total of 13,810 ML patients received autologous (n=9963) or allogeneic (n=3847) HCT between 1985 and 2012 were considered. At a median overall survival (OS) of 52 and 46 months in autologous and allogeneic HCT groups, respectively, ML patients receiving autologous HCT (1.38%; 95% confidence interval [CI], 1.09-1.68; at 3 year after autologous HCT) have a significant risk for developing t-AML/MDS compared to allogeneic HCT (0.37%; 95%CI, 0.06-0.67; at 3 year after allogeneic HCT). Median time from HCT to t-AML/MDS after autologous and allogeneic HCT was 957 and 414 days, respectively. Significant risk factors for the development of t-AML/MDS after autologous and allogeneic HCT are high-stage risk (P=0.04) or secondary malignancies (P Disclosures Iida: SymBio Pharmaceuticals: Research Funding. Suzumiya:Toyama Chemical: Research Funding; Kyowa Hakko kirin: Research Funding; Takeda: Honoraria; Eisai: Honoraria, Research Funding; Chugai: Honoraria, Research Funding; Astellas: Research Funding.

Introduction: CD5+ DLBCL comprises 5-10% of DLBCL and is an immunohistochemical subgroup of DLBCL, not otherwise specified. CD5+ DLBCL shows a significantly worse survival than CD5-negative DLBCL with rituximab (R)-CHOP. Of note, central nervous system (CNS) relapse, particularly brain parenchymal relapse, frequently occurs even in the R-era (Miyazaki K, et al. Ann Oncol 2011). Most CD5+ DLBCL is classified as non-germinal center B-cell-like (non-GCB)/activated B-cell-like DLBCL. DA-EPOCH-R is reported to show promising outcomes in patients with newly diagnosed non-GCB DLBCL (Wilson WH, et al. J Clin Oncol 2008). High-dose methotrexate (HD-MTX) therapy has a greater potential than intrathecal administration of MTX to prevent brain parenchymal relapse. To explore a more effective treatment for newly diagnosed CD5+ DLBCL, we conducted a multicenter phase II study of DA-EPOCH-R combined with HD-MTX (PEARL5 study; UMIN000008507). Methods: Patients with newly diagnosed stage II-IV disease from 20 to 75 years of age with ECOG performance status (PS) of 0 to 3 were eligible for enrollment. CD5 expression in tumor cells was examined by immunohistochemistry and/or flow cytometry in participating institutes. Four cycles of DA-EPOCH-R followed by 2 cycles of HD-MTX (3.5 g/m2) and additional 4 cycles of DA-EPOCH-R were planned as the protocol treatment. The primary endpoint was 2-year progression-free survival. Secondary endpoints were complete response (CR) rate, overall response rate, 2-year overall survival, 2-year CNS recurrence, and toxicity. Response was assessed at each participating institute using the revised International Workshop Criteria. Toxicity was graded according to the Common Terminology Criteria for Adverse Events v4.0. This planned interim analysis included response and toxicity by the protocol treatment. Results: Forty-seven patients were enrolled between Aug 2012 and Nov 2015. The diagnosis of CD5+ DLBCL in all patients was confirmed by the central pathology review. Cyclin D1 and EBER were negative in all cases examined (n = 47 and n = 46, respectively). According to the Hans' criteria, 72% (33/46) of the patients had non-GCB DLBCL and 28% (13/46) had germinal center B-cell-like DLBCL. The median age was 62 years (range: 37-74 years), and 60% were > 60 years. The baseline clinical features were as follows: female sex, 62%; ECOG PS >1, 4%; stage III/IV, 53%; elevated LDH, 66%; extranodal involvement > 1 site, 34%; bone marrow involvement, 11%; and high-intermediate-risk or high-risk groups of the International Prognostic Index, 47%. Skin/subcutaneous tissue was the most frequent site of extranodal involvement (15%). One patient had primary testicular DLBCL. Forty-six (99%) patients completed the protocol treatment. In the remaining patient, the protocol treatment was discontinued due to grade 3 stomatitis during the sixth cycle of DA-EPOCH-R. The median dose level of DA-EPOCH-R was 2 (range: 1-4). There was no deviation or violation in determining the dose levels of DA-EPOCH-R. At the time of this interim analysis, 46 patients were assessed for response. The CR rate was 91% (42/46; 95% confidence interval [CI], 79 - 98%) and the ORR was 93% (43/46; 95% CI, 82 - 99%). Two patients experienced disease progression during the treatment. Toxicity was assessed in 357 cycles of DA-EPOCH-R/HD-MTX in all 47 patients. There was no treatment-related death. Grade 4 non-hematologic toxicity was observed in only one patient who experienced tumor lysis syndrome with hyperkalemia soon after the first administration of R. The most common grade 3 non-hematologic toxicity was elevated alanine transaminase, which occurred in 6% of cycles. Two patients experienced grade 3 infection (cellulitis and endocarditis). Grade 3 neuropathy occurred in only 1% of cycles. There was no grade 3 cardiac toxicity. The targeted absolute neutrophil count (> 500/mm3) occurred in 77% of cycles. Thrombocytopenia of < 25,000/mm3 occurred in 8% of cycles. Febrile neutropenia (FN), which occurred in 23% of cycles, was manageable. Conclusion: These interim results demonstrate that DA-EPOCH-R/HD-MTX provides a high CR rate in patients with newly diagnosed stage II-IV CD5+ DLBCL. This therapeutic approach was feasible, although most patients experienced FN during the protocol treatment. Disclosures Miyazaki: Eisai: Honoraria; Kyowa Kirin: Honoraria; Chugai: Honoraria. Asano:Jannsen: Honoraria; Chugai: Honoraria. Nishikori:Janssen Pharmaceutical: Honoraria; Eisai: Honoraria, Research Funding; Kyowa Kirin: Honoraria. Sunami:Daiichi Sankyo: Research Funding; Novartis: Research Funding; Celgene: Honoraria, Research Funding; Janssen Pharmaceutical: Research Funding; Sanofi: Research Funding; Bristol-Myers Squibb K.K.: Research Funding; Takeda: Research Funding; Ono Pharmaceutical: Research Funding. Yoshida:Kyowa Kirin: Honoraria, Research Funding; Chugai: Honoraria, Research Funding; Celgene: Honoraria; Mundipharma: Membership on an entity's Board of Directors or advisory committees. Tamaru:Kyowa Kirin: Honoraria. Izutsu:Abbvie: Research Funding; Gilead: Research Funding; Celgene: Research Funding; Janssen Pharmaceutical K.K.: Honoraria; Eisai: Honoraria; Kyowa Hakko Kirin: Honoraria; Chugai Pharmaceutical: Honoraria, Research Funding; Takeda Pharmaceutical: Honoraria; Mundipharma KK: Research Funding. Kinoshita:Kyowa Kirin: Honoraria; Janssen: Honoraria; Solasia: Research Funding; Gilead: Research Funding; Takeda: Research Funding; Ono: Research Funding; Chugai: Honoraria, Research Funding; Zenyaku: Honoraria, Research Funding; Eisai: Honoraria, Research Funding. Suzumiya:Toyama Chemical: Research Funding; Takeda: Honoraria; Eisai: Honoraria, Research Funding; Chugai: Honoraria, Research Funding; Kyowa Hakko kirin: Research Funding; Astellas: Research Funding. Nishikawa:Daiichi Sankyo: Honoraria, Research Funding; Sanofi: Honoraria. Katayama:Takeda: Honoraria; Pfizer: Honoraria; Bristol-Myers Squibb Japan: Honoraria; Alexion Pharmaceuticals: Honoraria; Celgene: Honoraria; Chugai: Honoraria, Research Funding; Astellas: Honoraria, Research Funding; Daiichi Sankyo: Honoraria; Kyowa Hakko Kirin: Honoraria, Research Funding; Taisho Toyama Pharma: Honoraria; Shire: Honoraria; Nippon Shinyaku: Honoraria; Eisai: Honoraria; Dainippon Sumitomo Pharma: Honoraria; Shionogi: Honoraria. Yamaguchi:Zenyaku: Honoraria; Chugai: Honoraria; Kyowa Kirin: Honoraria; Takeda: Honoraria; Eisai: Honoraria.

Introduction: Diffuse large B-cell lymphoma (DLBCL) is the largest subtype of lymphoma, but consists of heterogeneous groups with different prognosis. Many studies conducted to determine prognosis factors of DLBCL. Recently, several studies have been reported that the elderly Epstein-Barr virus (EBV)-positive DLBCL patients showed worse prognosis than the elderly EBV-negative DLBCL. (Oyama T et al. Clin Cancer Res 2007;13:5024-5032). On the other hand, other studies reported that EBV-positivity is not a prognosis factor in young-adult DLBCL patients (Hong JY et al. Annals of Oncology 2015;26:548-555, Nicolae A et al. BLOOD 2015;126:863-872). In the present study, retrospective analyses were carried out for patients with DLBCL diagnosed between 1990 and 2007 to analyze the clinical and prognostic significance of EBV. Patients: Those who met the following criteria were included in the study: (i) pathology confirmed de novo DLBCL, according to the WHO classification; (ii) adequate amount and quality of paraffin-embedded biopsy specimens or unstained slides for EBV-encoded RNA in situ hybridization. Statistical methods: Intergroup comparisons were carried out with Fisher's exact test for categorical variables and the Mann-Whitney test for age. For survival analysis, we carried out Kaplan-Meier and multivariate proportional hazard (Cox) analyses. Statistical analyses were carried out using the software Stata SE version 14.0. Results: A total of 456 patients were included. The median follow up duration of survivor was 100 months (range, 1.9-363). There were 236 male (51.75%) and 220 female patients (48.25%) with a median age of 67 (range, 22-94) years. 246 patients (54.0%) were in advanced (III/IV) stage, 134 (29.4%) presented with poor Eastern Cooperative Oncology Group Performance Status (ECOG-PS), 275 (60.3%) with elevated lactate dehydrogenase (LDH) level, 263 (57.7%) with extranodal involvement of 1 or more sites, and 64 (14.0%) with bulky mass. As a consequence, 211 patients (46.3%) were in high/high-intermediate International Prognostic Index (IPI) categories. 5 years overall survival (OS) was 66.2%, in total. EBV was detected in samples from 47 patients (10.3 %). The positivity was 10.6% (15 / 142) in patients of 60 years or younger, and 10.2 % (32 / 314) in those older than 60 years (p = 0.51). No differences in back ground date were found between EBV - positive and negative DLBCLs regarding age (median 66.5 vs. 64.8, P = 0.17), male sex (88.1% vs. 11.9%, P = 0.16), advanced stage (88.2% vs. 11.8%, P = 0.17), poor ECOG-PS (90.3% vs. 9.7%, P = 0.47), elevated LDH (89.5% vs. 10.6%, P = 0.48), extranodal sites of one or more (90.5% vs. 9.5%, P = 0.31), bulky mass (92.2% vs. 7.8%, P = 0.33), and higher IPI categories (88.6% vs. 11.4%, P = 0.29). The prognosis was also not different between EBV-positive and negative DLBCLs. The 5 years OS was 42.6% for EBV-positive DLBCLs, and 56.4% for EBV-negative DLBCLs (P = 0.11). The same results were found for the younger (53.3% vs. 66.9%, P = 0.30) and the elderly patients (42.9% vs. 51.8%, P = 0.18). Conclusion: In this practice based, uncontrolled study, EBV-positively wad not different in younger and elderly DLBCLs. Although EBV-positive DLBCL showed worse trend of prognosis, the difference was not statistically significant. The significance of EBV in DLBCL should be reconsidered in unbiased, large-scale patient date. Figure Figure. Disclosures Suzuki: Chugai: Honoraria; Bristol-Myers Squibb: Honoraria; Kyowa Hakko kirin: Honoraria. Suzumiya:Chugai: Research Funding, Speakers Bureau; Astellas: Research Funding; Toyama Chemical: Research Funding; Eizai: Research Funding; Takeda: Speakers Bureau; Kyowa Hakko Kirin: Research Funding.

Background Natural killer (NK) cell malignancies are rare lymphoid neoplasms characterized by aggressive clinical behavior and poor treatment outcomes. Clinically they are classified as extranodal NK/T-cell lymphoma, nasal type (NKTCL) and aggressive NK cell leukemia (ANKL). Both subtypes are almost invariably associated with Epstein-Barr virus (EBV). Recently, genomic studies in NKTCL have identified recurrent somatic mutations in JAK-STAT pathway molecules STAT3 and STAT5b as well as in the RNA helicase gene DDX3X in addition to previously detected chromosomal aberrations. Here, we identified somatic mutations in 4 cases of ANKL in order to understand whether these entities share common alterations at the molecular level. To further establish common patterns of deregulated oncogenic signaling pathways operating in malignant NK cells, we performed drug sensitivity profiling using NK cell lines representing ANKL, NKTCL and other malignant NK cell proliferations. We aimed to identify sensitivities to agents that selectively target components of pathways required for survival of malignant NK cells in an unbiased manner. Methods Exome sequencing was performed on peripheral blood or bone marrow of ANKL patients using the NK cell negative fraction or other healthy tissue as control. Profiling of drug responses was performed with a high-throughput drug sensitivity and resistance testing (DSRT) platform comprising 461 approved and investigational oncology drugs. The NK cell lines KAI3, KHYG-1, NKL, NK-YS, NK-92, SNK-6 and YT and IL-2-stimulated and resting NK cells from healthy donors were used as sample material. All drugs were tested on a 384-well format in 5 different concentrations over a 10,000-fold concentration range for 72 h and cell viability was measured. A Drug Sensitivity Score (DSS) was calculated for each drug using normalized dose response curve values. Results The ANKL patients displayed mutations in genes reported as recurrently mutated in NKTCL, such as FAS, TP53, NRAS, STAT3 and DDX3X. Additionally, novel alterations in genes previously implicated in the pathogenesis of NKTCL were detected. These included an inactivating mutation in INPP5D (SHIP), a negative regulator of the PI3K/mTOR pathway and a missense mutation in PTPRK, a negative regulator of STAT3 activation. Interestingly, the total number of nonsilent somatic mutations in 3 out of 4 ANKL patients (97, 82 and 45) was remarkably high compared to other hematological malignancies analyzed in our variant calling pipeline. Analysis of drug sensitivities in NK cell lines showed a close correlation between all cell lines and a markedly higher correlation with those of IL-2 stimulated than resting healthy NK cells, suggesting that malignant NK cells may share a common drug response pattern. Furthermore, in an unsupervised hierarchical clustering the NK cell lines formed a distinct group from other leukemia cell lines tested (Fig. A). Among pathway-selective compounds (namely, kinase inhibitors and rapalogs), the drugs most selective for malignant NK cells fell into two major categories: PI3K/mTOR inhibitors (e.g. temsirolimus, buparlisib) and inhibitors of aurora and polo-like kinases such as rigosertib and GSK-461364 (Fig. B). JAK inhibitors (e.g. ruxolitinib, gandotinib) and CDK inhibitors (e.g. dinaciclib) showed strong efficacy in both malignant NK cells and IL-2 activated healthy NK cells. Conclusions Our exome sequencing results suggest that candidate driver alterations affecting similar signaling pathways underlie the pathogenesis of ANKL as has been reported in NKTCL. Drug sensitivity profiling highlights the PI3K/mTOR pathway as a potential major driver of malignant NK cell proliferation, whereas JAK-STAT signaling appears to be essential in both healthy and malignant NK cells. Components of these pathways harbored mutations in our small cohort of ANKL patients and have been shown to be deregulated by mutations or other mechanisms in previous studies, underlining their importance as putative drivers. The systematic large-scale characterization of drug responses also identified these pathways as potential targets for novel therapy strategies in NK cell malignancies. Figure 1. (A) Unsupervised hierarchical clustering based on drug sensitivity scores (DSS) of NK, AML, CML and T-ALL cell lines. (B) Scatter plot comparing DSS of malignant NK cell lines (average) and healthy IL-2 stimulated NK cells. Figure 1. (A) Unsupervised hierarchical clustering based on drug sensitivity scores (DSS) of NK, AML, CML and T-ALL cell lines. (B) Scatter plot comparing DSS of malignant NK cell lines (average) and healthy IL-2 stimulated NK cells. Disclosures Mustjoki: Novartis: Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria, Research Funding; Pfizer: Honoraria, Research Funding.

[Background and Objectives] The number of patients received allogeneic hematopoietic stem cell (HSC) transplantation is increasing year by year, particularly for elderly patients. Related donor is preferable than the unrelated, but the safety of elderly donors has not been clarified. For this purpose, the complications of elderly HSC donor were retrospectively analyzed in comparison with younger donors. [Materials and Methods] From September 2006 to December 2014, a total of 7,896 related HSC donations was reported to the Japan society for hematopoietic cell transplantation (JSHCT) registration system by 391 harvest teams. The day 30 check reports after harvest were available for 6,911 (87.5%) donations. Donors under 18 years old were excluded, and 6,297 donations were analyzed in the present study. For donor age analysis those ranging from 18 to 30 years were regarded as reference. The primary endpoint was the incidence of early severe adverse events (eSAEs) during the harvest or within the 30-day period. Statistical analyses were conducted using Fisher's exact test to compare the donor demographics and to identify the relationships between the incidence of eSAEs. Logistic regression analyses were used to analyze the factors influencing eSAEs. Data analyses were conducted using EZR software, version 1.23 (Saitama Medical Center, Jichi Medical University). This study was approved by the ethical committee of JSHCT and Shimane University. [Results] Median age of donors were 42 years old (range: 18-80). There were 3,232 male and 3,002 female donors. A total of 2,009 donations were planned to collect bone marrow (BM) and 4,288 donations were planned to collect peripheral blood stem cell (PBSC). Nine of the planned BM harvests and 64 of the planned PBSC collections were cancelled because of the various reasons including adverse reactions or poor stem cell mobilization. Three other donations planned to collect BM were changed to PBSC, and four donations planned to collect PBSC were changed to BM, vice versa. These altered cases were included to the analysis as an intention-to-harvest basis. Out of 6,297 HSC donations, 63 donors (1.0%) were reported by the harvest teams to have experienced eSAEs. SAEs were de\x{fb01}ned as follows: death, events dangerous to life, prolongation of hospitalization, permanent failure, disease or abnormality inherited to offspring and other important medical events. The details of eSAEs were pain (8), infection (8), allergy (6), blood access related (5), neuropathy (4), thrombocytopenia (4), liver dysfunction (2), gout (2), tetany (2), epidural hematoma (2), pulmonary embolism (1), and others (19). None died due to eSAEs and 81 percent of donors recovered from eSAEs in an average of 17 days. In comparison with reference (18 to 30 years) the relative risk of eSAEs was 0.74 for donors aged 31-35 years, 0.37 for 36-40 years, 0.62 for 41-45 years, 1.04 for 46-50 years, 0.77 for 51-55 years, 1.27 for 56-60 years, and 2.66 for 61-65 years. Those aged 61-65 years only had significantly elevated risk of eSAE by univariate analysis (P = 0.02). The incidence of eSAE in each age group are shown in the Table. Univariate logistic regression analysis also showed female sex (1.3% vs 0.7%, 95% CI 1.16-3.55, P = 0.01) and current health conditions (1.9% vs 0.9%, 95% CI 0.99-4.12, P = 0.04) were risk factors affecting eSAEs other than age. Donation type (bone marrow or peripheral blood), laboratory abnormality at health screening and JSHCT donor insurance eligibility criteria did not affect the eSAEs. Multivariate analysis revealed that age category of 61-65 years (Table) and female sex (OR 2.03, 95% CI 1.21-3.43, P = 0.01) were independent predictive factors. [Conclusion] The safety of elderly family HSC donors was significantly inferior to younger donors. Elderly family donors above age 61 should be selected carefully. These findings are useful for informed consent at donations of elderly family donors and consideration the upper limit of age of unrelated volunteer donors. Table 1. Relative risk of eSAE adjusted by sex Adjusted N incidence OR 95% CI P-value Total 1.0% 18-30 years 16/1493 1.1% 31-35 years 6/758 0.8% 0.75 0.29-1.91 0.54 36-40 years 3/756 0.4% 0.38 0.11-1.30 0.12 41-45 years 5/745 0.7% 0.62 0.23-1.71 0.36 46-50 years 8/720 1.1% 1.04 0.44-2.44 0.93 51-55 years 6/722 0.8% 0.77 0.30-1.98 0.59 56-60 years 10/739 1.4% 1.26 0.57-2.79 0.57 61-65 years 9/321 2.8% 2.66 1.16-6.06 0.02 66 years - 0/43 0.0% - - - Disclosures No relevant conflicts of interest to declare.

The majority of sinonasal non-Hodgkin's lymphomas (NHLs) are thought to originate from T-cell lineage. However, they often express natural killer (NK)-cell markers so that their origin still remains obscure. In this study, cell type of sinonasal NHLs were characterized by immunohistochemical and Southern blot analyses. We examined nine patients with sinonasal NHL. Six patients with tonsillar or pharyngeal non-B-cell lymphomas served as a control group. Immunohistochemical study showed that all nine cases of sinonasal NHL were CD56+CD2+, whereas controls were CD56-CD2+. According to the rearrangement of T- cell receptors (TCRs) and expression of CD3 markers, the sinonasal NHL cases were classified into three groups: TCR-CD56(Leu-19)+CD3(Leu4)- NHL (three patients), TCR-CD56+CD3+ NHL (five patients), and TCR+CD56+CD3+ NHL (one patient). In contrast, control patients' NHLs were TCR+CD56-CD3+. These results imply that eight cases of TCR-CD56+ sinonasal NHL are of NK-cell lineage. Among these eight cases, TCR- CD56+CD3+ cases (five of eight patients) were rather similar to the phenotype of fetal NK cells. From these results, the majority of sinonasal NHLs seem to originate from varying maturation stages of NK- cell lineage.

Epstein-Barr virus (EBV)–DNA was prospectively analyzed in plasma and mononuclear cells (MNCs) from peripheral blood in patients with extranodal natural killer (NK)/T-cell lymphoma, nasal type, to evaluate the clinical significance for diagnosis, monitoring the tumor burden, and prognostication. Thirty-three patients were enrolled, and 32 were evaluable. Pretreatment plasma and MNC EBV-DNA was detectable in 14 (range, 50-71 000 copies/mL) and 6 patients (range, 20-780 copies/μg DNA), respectively, and both were well correlated (r = 0.8741, P < .0001). Detectable plasma EBV-DNA was associated with higher clinical stage (P = .02), presence of B symptoms (P = .02), worse performance status (P = .02), and higher serum soluble IL-2 receptor level (P < .0001). Twenty-two patients attained complete response. Plasma EBV-DNA level was significantly higher in nonresponders than in responders (mean, 16 472 vs 2 645 copies/mL; P = .02). Multivariate analysis showed clinical stage (hazard ratio, 9.0; 95% confidence interval, 1.8%-45.0%) and pretreatment plasma EBV-DNA (hazard ratio, 10.6; 95% confidence interval, 1.3%-87.0%) were significant prognostic factors. Three-year overall survival of plasma EBV-DNA positive and negative patients was 42.9% and 94.4%, respectively (P = .0009). Plasma was a preferable sample for this purpose in NK/T-cell lymphoma, nasal type, and EBV-DNA level was a good indicator for response and overall survival.

CD10 and MUM1 are representative B cell differentiation markers. Follicular lymphoma (FL) is usually positive for CD10 and negative for MUM1. In this study, however, we compared 22 FLs with peculiar phenotype CD10−MUM1+ with 119 typical CD10+MUM1− FLs. All CD10−MUM1+ FL patients exhibited follicular structure with follicular dendritic meshwork, and a high rate of somatic hypermutation and ongoing mutation, similar to typical FL. However, CD10−MUM1+ FLs were encountered frequently in the elderly compared with CD10+MUM1− typical FLs (67.0 versus 58.7 years, P < .01), showed high grade (grade 3A or 3B) morphology (91% versus 17%, P < .001), diffuse proliferation (59% vs 19%, P < .001), and lacked BCL2/IGH translocation (5% versus 92.5%, P < .001), which is the most characteristic aberration in FL, and 88% showed BCL6 gene abnormalities (translocation or amplification). Our results indicate that CD10−MUM1+ FL is different from typical FL with respect to biologic and clinical features.

Introduction The outcome and the role of allogeneic hematopoietic cell transplantation (Allo-HCT) with reduced-intensity conditioning (RIC) in patients with nodal peripheral T-cell lymphomas (PTCLs) remain unclear. Patients and Methods To address this issue, we retrospectively analyzed the outcome of Allo-HCT for patients with nodal PTCLs using the transplant registry data from the Japan Society for Hematopoietic Cell Transplantation (JSHCT). Patients who fulfilled the following criteria were included in this study: aged 16-69 years, diagnosed with PTCL not otherwise specified (PTCL-NOS), angioimmunoblastic T-cell lymphoma (AITL), or anaplastic large cell lymphoma (ALCL), and received the first Allo-HCT in Japan between January 1, 2001 and December 31, 2011. In this analysis, conditioning regimen intensity was the main variable of interest. The conditioning regimen was classified as myeloablative conditioning (MAC) if it included total body irradiation (TBI) > 8 Gy, oral busulfan (BU) ≥ 9 mg/kg, intravenous BU ≥ 7.2 mg/kg, or melphalan (MEL) > 140 mg/m2. Otherwise, it was classified as RIC. Results A total of 354 patients (200 PTCL-NOS, 77 AITL, and 77 ALCL) were analyzed. Median follow-up duration of surviving patients was 3.8 years. Donor sources consisted of 122 human-leukocyte-antigen (HLA)-matched bone marrow (BM)/peripheral blood (PB), 122 HLA-mismatched BM/PB, and 110 cord blood. Of the 354 patients, 146 (41.2%) received MAC, which consisted of cyclophosphamide (CY)-TBI-based (n = 84), other TBI-based (n = 24), BU-CY-based (n = 11), fludarabine (FLU)-BU-based (n = 10), FLU-MEL-based (n = 15), and other (n = 2) MAC. The remaining 208 (58.8%) patients received RIC, which consisted of FLU-BU-based (n = 62), FLU-MEL-based (n = 108), and other (n = 38) RIC. Comparison of the patients who received MAC and RIC revealed that the RIC patients were significantly older (median age: 40.5 years vs. 50.3 years; P < 0.001) and more likely to have received autologous-HCT prior to Allo-HCT (15.1% vs. 29.3%; P = 0.002). The unadjusted 3-year cumulative incidence of non-relapse mortality were following: younger patients receiving MAC, 22%; younger patients receiving RIC, 14%; elderly patients receiving MAC, 50%; elderly patients receiving RIC, 30% (P < 0.001; Figure 1). The multivariate analysis showed that patients receiving RIC had a significantly lower non-relapse mortality than patents receiving MAC (HR, 0.51; 95% CI, 0.32-0.80; P = 0.004). Figure 1. Unadjusted non-relapse mortality. Figure 1. Unadjusted non-relapse mortality. The unadjusted 3-year cumulative incidence of relapse mortality were following: younger patients receiving MAC, 35%; younger patients receiving RIC, 30%; elderly patients receiving MAC, 32%; elderly patients receiving RIC, 35% (P = 0.692; Figure 2). The multivariate analysis showed that patients receiving MAC and RIC had a comparable relapse mortality (HR, 1.08; 95% CI, 0.73-1.58; P = 0.711). Figure 2. Unadjusted relapse mortality Figure 2. Unadjusted relapse mortality The unadjusted 3-year overall survival rates were following: MAC for younger patients (aged 16-49 years), 43%; RIC for younger patients, 56%; MAC for elderly patients (aged 50-69 years), 18%; RIC for elderly patients, 35% (P < 0.001; Figure 3). The multivariate analysis showed that patients receiving RIC had a significantly superior overall survival than patients receiving MAC (HR, 0.74; 95% CI, 0.54-1.00; P = 0.047; Table I). Figure 3. Unadjusted overall survival Figure 3. Unadjusted overall survival Table I. Multivariate analysis for overall survival Overall Survival HR (95% CI) P value Conditioning Regimen myeloablative 1.00 - reduced-intensity 0.74 (0.54-1.00) 0.047 Patient Age 16 to 34 1.00 - 35 to 49 1.26 (0.83-1.91) 0.270 50 to 59 2.17 (1.46-3.23) < 0.001 60 to 69 2.24 (1.40-3.59) 0.001 Karnofsly Performance Status 90 to 100 1.00 - 10 to 80 2.02 (1.41-2.91) Conclusion We showed a favorable outcome of Allo-HCT with RIC in patients with nodal PTCLs. The efficacy of RIC Allo-HCT for nodal PTCLs needs to be explored in prospective study. Disclosures No relevant conflicts of interest to declare.

Background: Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare malignancy of plasmacytoid dendritic cells as classified by the World Health Organization (WHO), and is characterized by distinct clinical, pathological and genetic features. BPDCN used to be diagnosed as blastic NK-cell lymphoma or CD4+/CD56+ hematodermic neoplasm. Its prognosis is extremely poor with a median overall survival (OS) of about 1 year. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) in remission has shown encouraging results for patients with BPDCN. However, the role of high-dose chemotherapy followed by autologous stem cell transplantation (HDT/ASCT) has not been reported. Methods: We surveyed the data of the Transplant Registry Unified Management Program of the Japan Society for Hematopoietic Cell Transplantation and identified 109 patients with an original diagnosis of BPDCN, blastic NK-cell or lymphoblastic lymphoma of NK-cell lineage. These diagnoses had been made by a pathologist at each institution in accordance with the WHO classification or previous lymphoma classification systems. Then, we sent out questionnaires to collect additional data on clinical presentation, prior therapies and update of follow-up, in addition to pathology and flow cytometry reports. The diagnosis of BPDCN was made with clinicopathological review by a consensus panel. Results: Of the 109 identified patients, additional data were obtained for 83. After central clinicopathological review, 58 patients were excluded from analysis due to disease other than BPDCN or the absence of important clinical data. Finally, the diagnosis of BPDCN was confirmed in 25 patients (allo-HSCT, N = 14; HDT/ASCT, N = 11). The median age at HSCT was 58 (range, 17-67) years and male patients were predominant (80%). Involvements of skin and bone marrow at diagnosis were observed in 88% and 68% of patients, respectively. The induction chemotherapy regimen was as follows: non-Hodgkin lymphoma (NHL)-like (e.g. cyclophosphamide, doxorubicin, vincristine, prednisone [CHOP]-like regimens; or ifosfamide/etoposide-based regimens) (N = 11), acute lymphoblastic leukemia (ALL)-like (N = 10) and acute myeloid leukemia (AML)-like (N = 4). The rate of initial response to induction chemotherapy was high, with a complete remission (CR) rate of 96%, except for one patient who experienced progressive disease in the central nervous system during induction therapy with an ALL-like regimen. The median time from diagnosis to HSCT was 6 months (range, 2-22). Among the patients who underwent allo-HSCT, myeloablative (MAC) and reduced-intensity (RIC) regimens were used in 8 (57%) and 6 (43%) patients, respectively. The donor of 1st allogeneic transplant was a relative in 7, unrelated bone marrow in 7 and cord blood in 1. Regarding the disease status at transplantation, all 11 patients underwent HDT/ASCT in CR1 whereas, among patients who underwent allo-HSCT, 12 were in CR1 (N=10)/CR2 (N =2) and two had disease. With a median follow-up of 53.5 months in surviving patients, OS and progression-free survival (PFS) at 4 years were 65% and 58%, respectively (Figure 1). The OS at 4 years for patients who received HDT/ASCT and allo-HSCT were 82% and 53%, respectively (P = 0.11, Figure 2). The PFS at 4 years for patients who received HDT/ASCT and allo-HSCT were 73% and 48%, respectively (P = 0.14, Figure 2). The three induction regimen groups (NHL-like, ALL-like and AML-like regimens) had similar OS rates: 62%, 70% and 67% at 4 years, respectively (P = 0.86). OS did not differ significantly between MAC and RIC groups (OS at 4 years: 45% vs. 60%, P = 0.31). There was no non-relapse mortality within 100 days after HSCT. Seven patients (28%) relapsed at a median of 10 months (range, 3-21) after HSCT (MAC allo-HSCT, N = 2; reduced-intensity conditioning allo-HSCT, N = 3; HDT/ASCT, N = 2). Notably, there was no relapse in patients without bone marrow (BM) infiltration at diagnosis in the HDT/ASCT treatment group. After allo-HSCT, grade 2-4 acute GVHD was observed in 5 patients (20%). Conclusions: As previous reports have shown, HSCT can achieve long-term survival in patients with BPDCN. In addition, the result of HDT/ASCT in CR1 seems to be promising and deserves further evaluation in the settings of prospective trials. Meanwhile, we need to clarify who could be cured by ASCT and need to improve outcome for high-risk patients, such as those with BM invasion. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Autologous or allogeneic hematopoietic stem cell transplantation (SCT) is considered in an attempt to improve survival in patients with relapsed or refractory lymphoma. The major problem with allogeneic transplantation is the increased treatment-related mortality (TRM) in comparison to autologous transplantation but some evidence supports the existence of a graft-versus-lymphoma (GVL) effect in lymphoblastic lymphoma with allogeneic SCT. Pediatric mature B-cell Non-Hodgkin lymphoma (B-NHL) is highly curable with intensive chemotherapy. It is hard to evaluate the role of autologous or allogeneic SCT in the era of modern chemotherapy regimens and the evidence for a potent GVL effect in B-NHL is not convincing. Therefore, we undertook a retrospective study of patients who underwent either autologous or allogeneic transplantation and analyzed the effect of several important factors, including donor source and survival rate, using the national registry data (Transplant Registry Unified Management Program; TRUMP). This study includes patients with B-NHL aged 18 years and younger who underwent single SCT facilitated by the TRUMP from 1990 to 2012. TRUMP system was established to unify of hematopoietic SCT registries in Japan. Primary end points were overall survival (OS), relapse, and TRM. TRM was defined as death during a continuous complete remission or death in the first 28 days after transplantation. There were 73 autologous transplantations and 36 allogeneic bone marrow transplantations available for analysis. Among the patients with autologous SCT, 8 underwent bone marrow transplantations and 65 underwent peripheral blood stem cell transplantations. Autologous transplant recipients were more likely to receive early transplants after diagnosis, more likely to receive transplants before 1999, less likely to be stage IV, and less likely to receive a TBI-containing conditioning regimen than allogeneic transplant recipients. With a median follow-up time of 5.9 years, the probabilities of OS were 57% (95% CI, 49%-66%) after allogeneic transplantation and 82% (95% CI, 78%-87%) after autologous transplantation (p = 0.004; Figure 1). Multivariate analysis confirmed that OS was significantly worse with allogeneic SCT (HR = 3.12, 95% CI, 1.01-9.69, p = 0.049), adjusted by the time from diagnosis to SCT, disease status at transplant, year of transplant, stage of lymphoma, and use of TBI. Cumulative incidence of relapse by type of transplant, showed no statistically significant difference after autologous versus allogeneic transplantation (p = 0.099; Figure 2). Seven patients died of transplant related complications including patients in the allogeneic SCT (n = 6) and autologous SCT (n = 1) groups. The causes of death were infection (n = 3), disease progression in the first 28 days after SCT (n = 3) and chronic graft-versus-host disease (n = 1). The 6-year cumulative incidences of TRM were 17% (95% CI, 6.8%-30%) after allogeneic transplantation and 1.4% (95% CI, 0.1%-6.6%) after autologous transplantation (p < 0.001, Figure3). There were too few events in the transplants to permit multivariate analysis. At every time point following transplantation, allogeneic transplant recipients were significantly more likely to die of treatment-related causes than autologous transplant recipients. In pediatric B-NHL, OS was significantly worse for allogeneic than for autologous SCT due to higher TRM with allogeneic SCT recipients. We have not found differences in the relapse rate between allogeneic and autologous SCT. Because TRM was the most important cause of treatment failure for the allogeneic patients, strategies to reduce TRM without compromising the GVL effect may be useful in pediatric B-NHL with SCT. Figure 1 Figure 1. Figure 2 Figure 2. Figure 3 Figure 3. Disclosures Fujita: Japan Leukemia Research Fund: Research Funding.

Introduction: Multiple myeloma is still incurable and optimize existing chemotherapeutic strategies and development of novel agents are necessary to improve the outcome of patients. Cotylenin A, a fusicoccane diterpene glycoside with a complex sugar moiety, was isolated as a plant-growth regulator. Cotylenin A modulates the 14-3-3 intracellular signaling pathway and has been shown to inhibit the growth of several cancer cells. Herein, we examined the antitumor effects of cotylenin A to develop a novel treatment against myeloma. Methods: Five human myeloma cell lines, RPMI8226, KMS-11, KMS-26, KMS-12 PE and KMS-12 BM were cultured with cotylenin A alone or in combination with several anticancer drugs. To measure the effects of various drugs on the growth of myeloma cells, the number of viable cells was determined by the MTT assay after 6 days of exposure to various concentrations of drugs with or without 2 μg/ml cotylenin A. The growth-inhibiting effects of the drugs were examined by determining the concentrations of drugs required to reduce the cell number to one-half of that in untreated cells (IC50). Xenografts in six-week-old female (Fox Chase SCID C.B-17/Icr-scid Jcl) mice were used to determine the in vivoefficacy of cotylenin A. Results: Cotylenin A alone inhibited the growth of myeloma cells in dose dependent manner, but the effect of growth inhibition was relatively weak. Cotylenin A and vincristine synergistically inhibited the growth and induced apoptosis in myeloma cells. While other microtubule-disturbing agents also showed co-operative effects with cotylenin A, other anticancer agents, such as doxorubicin, cisplatin, camptothecin, methotrexate, gemcitabine and 5-fluorouracil, did not show such co-operation with cotylenin A (Table 1). Cotylenin A had synergistic effects with vincristine and the results were confirmed by an isobologram analysis. These differences might be attributed to the effects on autophagic responses. Combined treatment with cotylenin A and vincristine induced autophagy (formation of LC3-II and degradation of p62 protein). However, doxorubicin did not enhance the autophagy induced by cotylenin A. The induction of apoptosis was confirmed by an analysis of the DNA histogram and the expression of annexin V. The expression of cleaved caspase-3 was increased in treatment with 2 ng/ml vincristine and enhanced by 2 μg/ml cotylenin A by western blot analysis. An invasion assay using transwell chamber revealed that low concentration cotylenin A (0.6 μg/ml) effectively inhibited the invasive activity of RPMI 8226 myeloma cells without inhibiting growth. A colony-forming assay indicated that 2 μg/ml cotylenin A preferentially inhibited the formation of large colonies, which have high self-renewal activity. The combined treatment with 3 μg/ml cotylenin A and 3 ng/ml vincristine more effectively suppressed the formation of large colonies than vincristine alone. Expression of pluripotency-associated transcription factor Sox2 mRNA in RPMI 8226 myeloma cells was significantly suppressed by treatment with 4 μg/ml cotylenin A. Combined treatment with 5 mg/kg cotylenin A and 0.5 mg/kg vincristine significantly inhibited the growth of KMS-26 myeloma cells as xenografts. In addition, mice with cotylenin A and vincristine showed the reduction of body weight loss comparing with those of mice by vincristine alone. This result supported that the combination of cotylenin A and vincristine might be safe. Conclusions: Cotylenin A and vincristine synergistically inhibited the growth of myeloma cells in vitro and in vivo, and the invasion of myeloma cells. Our results suggest that the combination of cotylenin A and vincristine may have therapeutic value for myeloma. Table 1. Potentiation of the growth-inhibitory activities of various anticancer agents in RPMI8226 myeloma cells by cotylenin A. Growth inhibition (IC50) Anticancer agent (ng/ml) - Cotylenin A + Cotylenin A Ratio(-/+) Doxorubicin 5.5 ± 0.6 5.3 ± 0.5 1.03 Cisplatin 246 ± 30.2 237 ± 28.4 1.03 Camptotecin 1.42 ± 0.16 1.38 ± 0.14 1.03 Methotrexate 2.76 ± 0.3 1.81 ± 0.2 1.52 Gemcitabine 4.3 ± 0.4 3.1 ± 0.3 1.39 5-Fluorouracil 56.5 ± 5.1 57.8 ± 6.3 0.98 Vincristine 6.3 ± 0.6 1.6 ± 0.1 3.94 Vinblastine 0.91 ± 0.11 0.42 ± 0.05 2.17 Paclitaxel 16.4 ± 1.9 7.7 ± 0.9 2.13 Ratio (-/+), IC50 without cotylenin A: IC50 with cotylenin A. The values are the mean ± SD of four determinations. Disclosures No relevant conflicts of interest to declare.

To the editor: We read with interest the results of the peripheral T-cell lymphoma (PTCL) classification project reported by Au et al, which stated that prognosis of extranodal natural killer (NK)/T-cell lymphoma (ENKL) of nasal origin is different from that of extranasal origin.[1][1] They further

Background Clinical impact of chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infections is not well studied in a large series of patients with lymphoma undergoing autologous stem cell transplantation (ASCT). Retrospective analysis was performed to evaluate clinical outcomes of patients with lymphoma receiving ASCT with or without HBV and HCV infections. Patients and Methods Among 13218 adult patients registered in the Adult Lymphoma Working Group of the Japan Society for Hematopoietic Cell Transplantation (JSHCT) database, we selected patients who had the information on HBV and HCV infection status and received ASCT from 1989 to 2010. Patients diagnosed as Hodgkin lymphoma, mature or precursor T- and B-cell lymphoma were included. Histological diagnosis of natural killer (NK)-cell lymphoma and adult T-cell leukemia/lymphoma were excluded in this analysis. Patients who had human immunodeficiency virus (HIV) infection and history of previous transplantation were also excluded. Survival analysis was performed to compare patients positive for HBV and HCV with those negative for both. Prognostic indicator for survival was also investigated in patients positive for HBV and HCV. Results A total of 4641 patients with 2819 male were analyzed. The median age of all patients was 54 years (range: 16-81 years). The patient characteristics were summarized in Table 1. HBV and HCV infections were present in 162 (3.5%) and 104 (2.2%) patients, respectively. Ten patients (0.2%) had both HBV and HCV infections. Detailed data on anti-HBV antigen or antibody and genetic data were not available in the analysis. Patients with HBV and HCV infections were older and more diagnosed with advanced stage than those without infections. With a median follow-up of 2.8 years, the 2-year overall survival (OS) rate of all patients was 75% (95%CI: 73 to 76). Cumulative incidence of treatment-related mortality (TRM) at one year after ASCT was 5.8% (95%CI: 5.2 to 6.6). According to HBV and HCV positivity, the estimated 2-year overall survival (OS) rates were 74% (95%CI: 65 to 81), 77% (95%CI: 66 to 84), 60% (95%CI: 25 to 83) and 75% (95%CI: 73 to 76) in patients with HBV-positive, HCV-positive, both virus positive and negative patients, respectively. Cumulative incidence of TRM at one year was 11% (95%CI: 6.2 to 16) 6.6% (95%CI: 2.7 to 13) and 5.7 (95%CI: 5.0 to 6.4) in patients with HBV-positive, HCV-positive and both virus negative patients, respectively. In patients with or without HBV and HCV infections, there was no statistically significant difference in rates of OS (p=0.82, Figure 1) and TRM (p=0.63). In multivariate analysis of patients infected with HBV, factors associated with shorter OS were male sex (HR: 3.1, 95%CI: 1.2 to 7.6), non-remission/relapse status at ASCT (HR: 2.7, 95%CI: 1.2 to 5.9) and ASCT before 2005 years (HR: 3.6, 95%CI: 1.6 to 8.4). ASCT before 2005 years was associated with higher TRM in patients with HBV infection (HR: 4.4, 95%CI: 1.5 to 13). In patients with HCV infection, multivariate analysis revealed that partial remission (HR: 3.5, 95%CI: 1.2 to 10.6), non-remission/relapse (HR: 5.3, 95%CI: 1.9 to 14.6) status at ASCT and age≥ 50 years at ASCT (HR: 7.0, 95%CI: 1.5 to 31.9) were significantly associated with shorter OS. Non-remission/relapse status at ASCT was associated with higher TRM in patients infected with HCV (HR: 4.4, 95%CI: 1.1 to 18.3). Difference in histology was an adverse factor for outcomes in the HBV- and HCV-negative groups, however, the factor could not be identified as adverse indicator in the HBV- and HCV-positive groups. Conclusions Prognosis of patients with HBV and HCV infections would be comparable to patients without those infections. Disease status at ASCT could be one of useful landmarks to predict outcomes in patients positive for HBV and HCV undergoing ASCT. After careful consideration, ASCT might be a feasible option for patients with HBV and HCV infections. Disclosures: No relevant conflicts of interest to declare.

Abstract 795 Background: Mogamulizumab (KW-0761) is a humanized anti-CCR4 antibody engineered to exert potent ADCC by defucosylation. In a phase I study for patients with CCR4-positive T-cell malignancies, once weekly administration for 4 weeks of mogamulizumab was well tolerated up to 1.0 mg/kg, and encouraging efficacy was observed (J Clin Oncol 2010;28:1591). In a subsequent phase II study in CCR4-positive relapsed adult T-cell leukemia-lymphoma (ATL) patients, mogamulizumab exhibited an overall response rate (ORR) of 50% (J Clin Oncol 2012;30:837), leading to its approval in Japan in 2012 for relapsed/refractory ATL. In addition, a phase I/IIa study for previously treated cutaneous T-cell lymphoma (CTCL) in the USA showed an ORR of 37% (14/38) (T-CELL LYMPHOMA FORUM 2012). Based on these findings, a phase II study of mogamulizumab for relapsed peripheral T-cell lymphoma (PTCL) and CTCL was conducted in Japan. Methods: A multicenter phase II study of mogamulizumab monotherapy for patients with relapsed CCR4-positive PTCL and CTCL was conducted to evaluate efficacy, pharmacokinetic profile, and safety. The primary endpoint was ORR and secondary endpoints included progression-free survival (PFS) and overall survival (OS). At least 35 patients were needed to detect a lower limit of the 95% confidence interval (CI) exceeding the 5% threshold, with an expected ORR for mogamulizumab of 25% with 90% statistical power. Patients received intravenous infusions of mogamulizumab once per week for 8 weeks at a dose of 1.0 mg/kg. Responses were assessed after the 4th and 8th infusions of mogamulizumab by an independent efficacy assessment committee. The histopathological subtypes of PTCL were confirmed by an independent pathology review committee according to the 2008 WHO classification. In addition, we examined blood T-cell subset distributions. Results: A total of 38 patients were enrolled, and 37 patients (male/female 23/14; median age 64 years, range 33–80) received mogamulizumab. One patient was withdrawn due to an infectious complication. Twenty-nine of the 37 assessable patients had PTCL [PTCL- not otherwise specified (NOS), n=16; angioimmunoblastic T-cell lymphoma (AITL), n=12; anaplastic large cell lymphoma (ALCL)-ALK negative, n=1] and 8 had CTCL [mycosis fungoides (MF), n=7; cutaneous ALCL, n=1]. Performance status at enrollment was 0 (n=24), 1 (n=12), and 2 (n=1). The median number of prior systemic chemotherapy regimens was 2 (range 1–6). Of the 37 patients, 25 completed the schedule of 8 planned infusions. Nine patients (24%) discontinued the treatment protocol due to progressive disease and 3 due to adverse events (AEs). The ORR in 37 patients was 35% (13/37, 95% CI, 20 to 53%) with 14% having a complete response (5/37) (Table 1). By PTCL subtype, the ORR was 34% (10/29) for PTCL (3/16 for PTCL-NOS, 6/12 for AITL, and 1/1 for ALCL-ALK negative) and 38% (3/8) for CTCL (2/7 for MF and 1/1 for cutaneous ALCL). AEs possibly, probably, or definitely related to mogamulizumab monotherapy were as follows. Lymphopenia of all grades and that of grades 3–4 were observed in 78% and 70% of the 37 patients, respectively. Leukopenia of all grades and that of grades 3–4 were observed in 43% and 14% of the 37 patients. For all grades and grades 3–4, neutropenia was observed in 35% and 16%, thrombocytopenia in 35% and 3%, ALT increases in 22% and 3%, and skin eruptions in 49% and 8% of patients, respectively. Infusion-related toxicities occurred in 22%, which were all within grade 2 or lower. Fourteen severe AEs were observed in 7 patients, including a grade 3 polymyositis in 1 and grade 2 cytomegalovirus retinitis in 2. All severe AEs were improved. No grade 5 AEs were observed. Pharmacokinetic analysis demonstrated that Cmax and trough (C168h) after the 8th infusion were 45.9 ± 9.3 and 29.0 ± 13.3 μg/mL, respectively. No anti-mogamulizumab antibody has been detected. Updated results of PFS, OS, and T-cell subset analysis are being analyzed for presentation. Conclusions: Mogamulizumab monotherapy showed promising antitumor activity with acceptable toxicity profiles in patients with relapsed PTCL and CTCL, warranting further investigation. Disclosures: Ishida: Kyowa Hakko Kirin Co., Ltd,: Honoraria, Research Funding, Speakers Bureau. Ogura:Kyowa Hakko Kirin Co., Ltd,: Consultancy. Suzumiya:Kyowa Hakko Kirin Co., Ltd,: Consultancy. Inagaki:Kyowa Hakko Kirin Co., Ltd,: Consultancy. Tamura:Kyowa Hakko Kirin Co., Ltd,: Consultancy. Akinaga:Kyowa Hakko Kirin Co., Ltd,: Employment. Tomonaga:Kyowa Hakko Kirin Co., Ltd,: Consultancy. Ueda:Kyowa Hakko Kirin Co., Ltd,: endowed chair Other.

Abstract 4594 Background: B-chronic lymphocytic leukemia (B-CLL) is rare in Japan. Therefore, only a few data are available on the clinical characteristics of B-CLL for Japanese patients (pts). We reported that the rate of atypical CLL appeared to be higher in Japanese pts than that of Caucasians. Patients and Methods: This nation-wide study was performed from Oct. 2003 to March 2008, and was supported by Japanese Elderly Leukemia and Lymphoma Study Group. The study was conducted in accordance with the Declaration of Helsinki and was approved by the Institutional Review Boards of the core members' hospitals. The pts with CLL and CLL-like disease were registered. Precursor leukemia and adult T-cell leukemia/lymphoma were excluded. The smears of peripheral blood and bone marrow of all the registered pts were examined by the Diagnosis Committee by using the 4th edition of WHO classification and FAB classification. FAB classification defines typical CLL as prolymphocytes≤10% and atypical CLL including CLL/PLL as 11%≤prolymphocyte15%. Mantle cell lymphoma was excluded by means of cytogenetics, fluorescent in situ hybridization of CCND1-IGH, and immunohistochemistry of CCND1. Results: One hundred and thirty-one evaluable pts were registered. There were 96 pts with B-CLL, 2 SLL, 1 B-PLL, 3 hairy cell leukemia, 16 leukemic phase of other B-cell lymphomas. Thirteen pts had NK or T cell leukemia/lymphoma with 7 T-LGL, 4 T-PLL, 1 NK-LGL, and 1 Sezary syndrome. Among B-CLL, there were 62 pts with typical morphology (typical CLL), 18 with atypical morphology (atypical CLL), 16 with intermediate morphology between typical and atypical CLL (intermediate CLL). The differences among 3 subgroups are shown in Table. No differences were found between the 3 morphological subgroups for age, sex, stage (modified Rai and Binet classification), hemoglobin level, platelet counts, the presence of hepatomegaly, splenomegaly and lympoadenopathy, ECOG-PS and the rate of treatment requirement. However, the incidence of CD5 and CD23 expression was higher in typical CLL than in atypical/intermediate morphology groups (P Conclusion: Morphological atypia was more frequent in Japanese B-CLL pts, partly because of the low incidence of typical CLL. The atypical/intermediate CLLs were associated with atypical immunophenotype (absence of CD5 and/or CD23) and favorable prognosis. Prospective study is ongoing to evaluate other prognositic factors including the implication of IgVH gene mutation status and ZAP-70 expression in Japan. Disclosures: No relevant conflicts of interest to declare.

Abstract 503 Background: Extranodal NK/T-cell lymphoma, nasal type (ENKL) is a rare subtype of lymphoma and its standard therapy is not established. The response to conventional chemotherapy is not good, resulting in generally poor prognosis. Hematopoietic stem cell transplantation (HSCT) is a promising strategy, but no large-series studies were presented. Methods: The Lymphoma Working Group of JSHCT surveyed the database in Japan and retrospectively analyzed. Of 11,267 transplants for all types of lymphoproliferative disorders, a total of 134 HSCT were conducted for 120 ENKL patients with sufficient data for analysis. The study was performed in accordance with the Declaration of Helsinki and was approved by the JSHCT Ethical Committee and the Institutional Review Boards of Nagoya University. Results: There were 75 male and 45 female with a median age of 47 years (range: 18–70). The median follow-up of survivor was 27 months. In total, 74 patients received allogeneic hematopoietic stem cell transplantation (60 with 1st transplant and 14 with 2nd transplant) and 60 received autologous peripheral blood stem cell transplantation (PBSCT). The donor of allogeneic 1st transplant was relatives in 28, unrelated bone marrow (UR-BM) in 9, and cord blood (CB) in 23. For 1st transplant, 63 patients were in complete response (CR) (41 autologous, 10 relative, 5 UR-BM, and 7 CB), 12 were in partial response (PR) (5 autologous, 4 relative, 1 UR-BM, and 2 CB), and 45 were on disease (14 autologous, 13 relative, 4 UR-BM, and 14 CB). For patients who received allogeneic transplant, 38 (64%) were in stage IV at initial presentation, while only 20 (33%) were in stage IV for patients who received auto-transplant. The proportion of low international prognostic index was also different between allo-transplant (34%) and auto-transplant (62%) groups. The 2-year overall survival (OS) was significantly better for those received auto-transplant than allo-transplant (69% vs. 41%, Figure 1). The OS of patients who received 2nd transplant was 27% at a median follow-up of 18 months. Multivariate analysis showed that three factors (initial stage IV, non-CR status and performance status at transplant) were significant factors associated with poor prognosis, but the type of transplant (autologous or allogeneic) was not. By adjusting above three factors, the difference between autologous and allogeneic transplant was minimized (Figure 2). Conclusion: Prognosis of ENKL patients who received autologous transplant was better than that of allogeneic transplant, partly due to the selection that patients in good condition tended to receive auto-transplant. In another words, for patients with good condition, autologous HSCT is a good treatment choice. Any type of HSCT is a potential option for ENKL patients in non-CR. The 2nd SCT is beneficial for a part of patients. Disclosures: No relevant conflicts of interest to declare.

Abstract 5137 [Background] Bortezomib and doxorubicin have synergistic activity against myeloma cells. However, the combination of bortezomib and cytotoxic agents has not long been permitted in Japan because the safety of the combination therapy has not been determined in Japanese patients. We, therefore, decided to undergo a phase I/II study of bortezomib, doxorubicin and intermediate-dose dexamethasone (iPAD) combination therapy in Japanese patients with relapsed or refractory myeloma. [Methods] In a phase I study, the maximally tolerated dose of bortezomib was determined when given in combination with a fixed dose of doxorubicin and dexamethasone. Bortezomib was administered iv on days 1, 4, 8 and 11 at a dose of 1.0 mg/m2 in cohort 1 and 1.3 mg/m2 in cohort 2. Doxorubicin 9 mg/m2 was given iv on days 1–4, and dexamethasone 20 mg po on days 1–2, 4–5, 8–9 and 11–12. After a recommended dose of bortezomib was determined, a phase II study was undertaken to examine overall response rate, survival time and toxicity profiles. [Results] The dose-limiting toxicity (DLT), defined as grade 4 hematological toxicity lasting more than 5 days and/or grade 3 or higher non-hematological toxicity, was observed in 2 of 6 patients in cohort 1, indicating this dose was considered tolerable. In cohort 2, 3 of 5 patients developed DLTs including grade 4 thrombocytopenia lasting more than 5 days, grade 3 elevated hepatic transaminase levels and grade 3 ileus in one of each patient, indicating this dose was intolerable. Thus, 1.0 mg/m2 of bortezomib is a recommended dose in iPAD therapy. In a phase II study, 27 patients, aged 40 to 78 (median 63), were enrolled to receive iPAD therapy by using 1.0 mg/m2 dose of bortezomib. Overall response (CR+PR) was achieved in 89% (95% CI, 76–100%), and complete response (CR+nCR) in 30% (95% CI, 11–48%). The median overall survival time was not reached and progression-free survival time was 12.3 months (95% CI, 6.1–18.5 months). One patient died of pneumonia. Grade 3/4 thrombocytopenia, neutropenia and anemia developed in 67%, 41% and 19%, respectively, and febrile neutropenia in 11%. Sensory neuropathy was observed in 78% with grade 3/4 in 22% of the patients. Grade 3/4 hypokalemia developed in 15%, hyponatremia in 11%, and fatigue, nausea and diarrhea in 4%. [Conclusion] The iPAD therapy is feasible and effective to treat relapsed or refractory myeloma in Japanese population. (This clinical study was registered in University hospital Medical Information Network (UMIN) (registration number; UMIN000001210)) Disclosures: No relevant conflicts of interest to declare.

Abstract 3093 Allogeneic hematopoietic stem cell transplantation (allo SCT) is potentially curative treatment for relapsed follicular lymphoma (FL) that remains non-curative disease even with modern immunochemotherapy. Number of potential candidate of allo SCT for FL has been increasing with the development of reduced intensity conditioning (RIC) regimens and increased donor availability due to use of hematopoietic stem cell from unrelated donor including cord blood (CB). However, short-term and long-term transplantation-related complications are still major obstacles in applying this treatment. Recently, EBMT and CIBMTR reported a prognostic score of allo SCT for FL (Ann Oncol 2011;22 :Suppl 4, iv94). However, this score awaits validation. To elucidate prognostic factors for OS after allo SCT for FL, we conducted a retrospective study using the national registry data of the Japan Society of Hematopoietic Cell Transplantation (JSHCT). In total, 472 cases of allo SCT for FL performed from 2000 to 2008 were identified. 220 (46.6%) were male and the median age at allo SCT was 50 yo (range: 27–75). Forty six (9.7%) patients were 60 yo or older. Eighty two (17.3%) patients had previous history of autologous transplantation (ASCT). Stem cell source was BM or PB from related donor in 215 (45.5%), unrelated donor BM in 180 (38.1%), and unrelated donor umbilical cord blood (CB) in 77 (16.3%). Conditioning regimen was myeloablative (MAC) in 20.7% and reduced intensity (RIC) in 79.3%, respectively. Patients undergoing MAC were younger than those with RIC (45 vs 50 yo, P Disclosures: No relevant conflicts of interest to declare.

Abstract 3091 Background: Aggressive NK cell leukemia (ANKL) is a malignant disorder of mature NK cells that is more common in East Asia and is often associated with Epstein-Barr virus (EBV). The prognosis of ANKL is dismal and the survival is one of the shortest among the lymphoid neoplasms. Because of its rarity, the characteristics and optimal managements of this disease are uncertain. To better elucidate clinicopathologic features and therapeutic modalities for ANKL, a retrospective Japan-Korea multicenter survey of ANKL (ANKL07) was conducted. Methods: Eligibility criteria were based on the diagnosis for ANKL according to the WHO classification. Pathological and clinical information including that on chemotherapy and hematopoietic cell transplantation (HCT) was collected. Approval from the institutional review board at each participating institution was obtained for this study. Results: Forty-one patients (pts) were registered from 8 hospitals. After central reviews, 34 pts were further analyzed, while 7 pts were excluded. The median age of the pts was 45 years old (range: 16–79) with 26 males and 8 females. Four pts had a history of preceding disorders including EBV-LPD in childhood, mosquito bite hypersensitivity or EBV-associated liver damage. Three female pts presented during pregnancy. Fever and hepatosplenomegaly were recognized in 100% and 71% of the pts, respectively. Thirty-two pts (94%) showed high or high-intermediate International Prognosis Index. Three types of ANKL cells (type I, II or III) were categorized according to the morphological features. Using this classification, there were 11, 13 and 10 pts with types I, II and III, respectively. EBV was positive in 85% of the pts. Eleven pts showed less than 20% leukemic cells in both peripheral blood (PB) and bone marrow (BM). The clinical characteristics and prognosis of these pts did not differ from those of the pts with more tumor cells in PB/BM except for the incidence of hemophagocytic syndrome. As initial therapy, anthracycline-based chemotherapies were utilized in 13 pts and L-asparaginase-containing regimens were administered in 5 pts. Although anthracycline chemotherapy was not associated with better outcome by Kaplan-Meier analysis (p=0.64), L-asparaginase chemotherapy resulted in CR or PR in 4 pts and significantly better survival (p=0.03). A total of 9 HCT, 2 autologous (auto) and 7 allogeneic (allo), were performed in 8 pts; none were in complete remission (CR) at the time of transplant. For allo HCT, the donor source was HLA-matched related bone marrow in two, cord blood in two, PB of HLA-matched unrelated donor in one and HLA-mismatched related PB in two. The conditioning regimen was TBI+CY in three, and fluradabine-based regimen in four. One patient with auto HCT and 4 with allo HCT reached CR after HCT. The median survival of all 34 pts was 51 days (range: 1-1, 630). Median survivals for the patients with or without HCT were 266 days (range: 80-1, 630) and 36 days (range: 1–324), respectively. Two pts with allo HCT were alive in CR while the other 6 pts with allo HCT died of the disease (5 pts) or infection (1 pt). All pts without HCT died of ANKL. Times from the diagnosis to HCT for the 2 survivors were within 80 days. In univariate analysis, L-asparaginase administration was the only clinical factor associated with better survival (hazard ratio 0.29, 95% confidence interval: 0.12–0.70, p=0.008). Age (HR 1.02, 95% CI: 0.998–1.08, p=0.07) and the use of etoposide (HR 0.48, 95% CI: 0.22–1.07, p=0.07) were marginal. Multivariate Cox analysis indicated that the use of L-asparaginase was an independently significant factor for better survival (HR 0.33, 95% CI: 0.13–0.83, p=0.02). Conclusions: This is the largest case series of ANKL. ANKL cells are morphologically heterogeneous and the main infiltration sites of ANKL are liver, spleen and BM. In addition, ANKL showed poor prognosis regardless of PB/BM tumor cell burden. Although the prognosis of ANKL did not improve significantly compared with those in previous reports, L-asparaginase-containing initial chemotherapy and allo HCT are promising approaches for ANKL. Prospective studies are required to confirm these results. Disclosures: No relevant conflicts of interest to declare.

Abstract 2873 Background: Extranodal NK/T-cell lymphoma, nasal type (ENKL) is a rare subtype of lymphoma and its standard therapy is not established. Because of the expression of multi-drug resistance associated P-glycoprotein, CHOP (-like) chemotherapy has limited efficacy in ENKL, with an overall response rate (ORR) of 36% for newly-diagnosed stage IV diseases and less than 10% for relapsed or refractory diseases. Our previous phase I trial of a new chemotherapeutic regimen, SMILE (Steroid=dexamethasone, Methotrexate, Ifosfamide, L-asparaginase and Etoposide) showed promising results (Cancer Sci, 2008). ENKL is invariably associated with clonal episomal infection of Epstein-Barr virus (EBV) in the lymphoma cells. The peripheral blood of these patients contains fragmented EBV-DNA, which can be used as a quantitative surrogate tumor marker for disease diagnosis, monitoring and prognostication. Methods: A phase II study of SMILE chemotherapy was conducted. Patients (Pts) with newly-diagnosed stage IV, or relapsed/refractory disease after first-line chemotherapy, between 15–69 years old and with a PS of 0–2 were eligible. Primary endpoint was ORR after 2 cycles of SMILE chemotherapy, and target enrollment was 38 pts. To ameliorate myelotoxicity and based on the results of phase I study, G-CSF was started from day 6. A part of this study (anti-tumor effect of the SMILE chemotherapy) has been presented in ASCO 2010 (abstract #8044) and EHA 2010 (abstract #299) meetings. EBV-DNA copy number before treatment was analyzed by using quantitative polymerase chain reaction. In Japan, the EBV-DNA was measured at the central laboratory (Nagoya Univ.). In Hong Kong and Korea, it was measured in each country, and was converted using coefficients. The association of converted EBV-DNA and anti-tumor effect of SMILE chemotherapy was investigated. Results: From July 2007 to October 2009, a total of 39 pts were enrolled in the phase II SMILE study. The median age was 47 years (range: 16–67), and male/female ratio was 21/18. There were newly-diagnosed stage IV disease in 21, first relapse in 13, and primary refractory disease in 5 pts. EBV-DNA before treatment was measured in 38 patients using whole blood (N=34) or plasma/serum (N=33). In 29 patients the EBV-DNA was examined in both samples. The median EBV-DNA was 2,850 copies/mL (range: 0–1.14×10^7) in whole blood and 1,101 copies/mL (range: 0–1.27×10^7). Results of these 2 measurement of EBV-DNA well correlated (r = 0.8706, P < 0.0001). EBV-DNA was undetectable (below the cutoff level) in 7 of 34 patients using whole blood and 14 of 33 patients using plasma/serum. 29 pts (74%) completed the planned treatment. The responses were complete remission (CR) in 15, partial remission (PR) in 14, no response in 4, progressive disease in 3, and early death in 3. ORR and %CR were 74% (90% confidence interval, 60–85) and 38%, respectively. Grade 4 neutropenia occurred in 36 of 39 patients (92%), and 2 patients died of infection during neutropenia. Grade 4 non-hematologic toxicities were infection (n=5), hyperbilirubinemia (n=1), ALT elevation (n=2) and encephalopathy (n=1). The grade 4 non-hematologic toxicity was only experienced in the 1st course of SMILE. No patient developed grade 4 non-hematologic toxicity in the 2nd course. The most common grade 3 non-hematologic toxicity was infection (41%). The whole blood EBV-DNA copy number was associated with ORR (r=0.71) and %CR (r=0.78). ORR was 88% in patients with less than 10^5 copies/mL EBV-DNA in whole blood, but was 44% in patients with more than 10^5 copies/mL (P=0.02). Six of 7 patients without detectable EBV-DNA in whole blood attained CR. In contrast, none of the 8 patients with more than 10^5 copies/mL EBV-DNA could achieve CR. Grade 4 non-hematologic toxicity was significantly higher in patients with more than 10^4 copies/mL of EBV-DNA in plasma (55% vs. 14%, P=0.03). Conclusion: Our results indicate that SMILE chemotherapy is an effective treatment for newly-diagnosed stage IV, relapsed or refractory ENKL. Myelosuppression and infection during the treatment should carefully be managed. EBV-DNA copy number is also predictive for response and adverse events of SMILE chemotherapy for newly-diagnosed stage IV, relapsed or refractory ENKL. Disclosures: Suzuki: Kyowa Kirin Company: Honoraria. Oshimi:Eisai Pharmaceutical Co Ltd: Employment.

Abstract 135 Background: Peripheral blood of patients with extranodal NK/T-cell lymphoma, nasal type (ENKL) contains fragmented Epstein-Barr virus (EBV) DNA. Measurement of the circulating viral DNA load has been reported to be useful for the diagnosis, monitoring and prognostication of the disease. However, there are two different subjects for analysis, plasma/serum component and mononuclear cells (MNC). Most reports do not compare both samples from the same patients, and are biased by a retrospective manner of accrual. Therefore, it remains unclear which samples are more useful subjects. Materials and Methods: To evaluate clinical significance of peripheral blood EBV-DNA copy number for ENKL, we conducted a prospective study to analyze EBV-DNA with quantitative polymerase chain reaction. Inclusion criteria are as follows: (1) Diagnosis of ENKL or aggressive NK-cell leukemia by biopsy or cytology. (2) Patients without other serious complications and those tolerable for chemotherapy and/or radiotherapy. (3) No prior history of chemotherapy or radiotherapy. (4) Patients with written informed consent. Primary endpoint was a prognostic value of EBV-DNA copy number to predict 2-year overall survival. Secondary endpoints were comparison of EBV-DNA copy number and pretreatment characteristics and prognostic capability of EBV-DNA during/after treatments. Three times of analysis, pre-treatment, mid-treatment, post-treatment, at central laboratory was to be performed for each patient. Results: A total of 33 patients were registered from June 2004 to March 2007. All patients were diagnosed with ENKL and male/female ratio was 21/12. The median age was 56 years old, ranging from 18 to 81. 19 patients were in stage IE, 3 in stage IIE, and 11 in stage IV. 13 patients had B-symptom and 14 had elevated serum LDH level. ECOG performance status was low (0–2) in 29 patients, but was 3 in 2 patients. IPI was high-intermediate/high in 11. First line treatment included concurrent chemoradiotherapy in 16, radiotherapy followed by chemotherapy in 8, and chemotherapy alone in 6. Pretreatment MNC and plasma EBV-DNA were detectable in 6 and 14 patients, respectively. The maximum copy numbers were 780 copies/microgram DNA for MNC and 71000 copies/mL for plasma. Significant correlation was observed between mononuclear and plasma EBV-DNA copies (r = 0.8741, P < 0.0001). Plasma EBV-DNA well correlated with pretreatment clinical stage (P = 0.02), presence of B-symptom (P = 0.02), ECOG PS (P = 0.02), serum LDH level (P = 0.05), and soluble IL-2 receptor (P < 0.0001), but not with regional node involvement (P = 0.17), nasal vs. nasal-type (P = 0.16), and serum C-reactive protein (P = 0.29). Among 28 patients evaluable for response, 21 patients responded (CR/PR) to the first line treatment. Mean plasma EBV-DNA copy number before treatment was significantly higher in non-responders compared to responders (16472 copies/mL vs 2645 copies/mL, P = 0.02). 2-year overall survival was 69.7%. The median follow-up of patients was 2.9 years. Clinical stage, performance status, pretreatment plasma EBV-DNA and pretreatment mononuclear cell EBV-DNA were significant prognostic factors for overall survival by univariate analysis. Multivariate analysis showed clinical stage (hazard ratio = 9.0, 95% confidence interval: 1.8–45.0) and pretreatment plasma EBV-DNA (hazard ratio = 10.6, 95% confidence interval: 1.3–87.0) were significant prognostic factors. 2-year overall survival of plasma EBV-DNA positive and negative patients was 42.9% and 94.4%, respectively (Figure, P = 0.0009). Conclusions: Our study shows pretreatment plasma EBV-DNA copy number is a good indicator for both response to treatment and overall survival. Measurement of the plasma EBV-DNA is useful for prospective clinical trials and general practice. Disclosures: Oshimi: Eisai Pharmaceutical Company: Employment.

Abstract 4812 Gemtuzumab ozogamicin (GO) is an immunoconjugate of an anti-CD33 antibody and a toxic calicheamicin-γ1 derivative that facilitates cellular uptake of the latter drug. CD33 is selectively expressed on myeloid cells, especially on acute myeloid leukemia (AML) cells, but usually not on non-hematopoietic cells, and therefore the clinical use is confined to AML. Despite its predominant expression on AML cells, GO induces complete remission only in 30% of recurrent AML. The major side-effects of GO include not only myelosuppression but also hepatotoxicity, although CD33 is not expressed on hepatobiliary cells. It suggests that the efficacy of GO is not strictly regulated by CD33 expression. We investigated relationship between CD33 expression and GO cytotoxicity by using leukemia cell lines, and classified them into four groups; group A defined as CD33(+) and cytotoxicity (+), group B as CD33(+) and cytotoxicity (-), group C as CD33(-) and cytotoxicity (+), and group D as CD33(-) and cytotoxicity (-). We selected four cell lines, HL-60, MEG-01, MOLT-3 and K562 which belong to group A, B, C and D, respectively, and were used for further experiment. CD33 was expressed on HL-60 but not on MOLT-3. However, GO-induced cytotoxicity was seen in both cells. We therefore compared effects of GO in HL-60 and MOLT-3. When incubated with GO for 72 hours, viability of HL-60 significantly decreased at 10 ng/mL of GO, while cytotoxic effect of GO against MOLT-3 became apparent when GO concentration was raised to 100 ng/mL. Viability of HL-60 markedly decreased when treated with GO for 30 minutes and then cultured in medium without containing GO for 72 hours, but no cytotoxic effect was observed in MOLT-3 unless culture with GO was prolonged to more than 6 hours. GO-induced apoptosis was detected in both HL-60 and MOLT-3 by Annexin V assay and TUNEL assay. The pretreatment with unconjugated anti-CD33 antibody (hP67.6) resulted in reduced cytotoxic effects of GO in HL-60, but not in MOLT-3. It indicates that GO induces apoptosis in CD33-non-expressing MOLT-3. Higher doses and longer exposure of GO treatment are necessary to induce cytotoxicity against MOLT-3 when compared with HL-60. GO might cause cytotoxicity against MOLT-3 through drug absorption pathway other than CD33-mediated endocytosis. Although HL-60 and MEG-01 expressed CD33, GO induced cytotoxicity on HL-60, but not on MEG-01. We then investigated internalization of antibody-bound CD33. After incubated with hP67.6 on ice, cells were cultured in antibody-free medium, and remaining cell surface-associated CD33 was measured. The loss of cell surface CD33 was observed in MEG-01 as well as HL-60, showing that internalization of CD33 following antibody binding occurred in MEG-01. It indicates that resistance to GO-induced cytotoxicity in MEG-01 is not associated with internalization property. Drug transport pathway other than receptor-mediated endocytosis was investigated. Cells were cultured in rhodamine 123 (Rh123)-containing medium for 30 minutes and the influx of Rh123 was assessed by the fluorescence activity of Rh123-containing cells. Cells were then cultured in medium without containing Rh123 for further 3 hours, and the efflux of Rh123 was analyzed by measuring the reduction curve of fluorescence activity. Rh123 accumulated in MEG-01 twice higher than HL-60. However, fluorescence activity of Rh123 decreased to 20% after 3 hours, while it remained unchanged in HL-60, indicating that drug efflux was activated in MEG-01. Multidrug resistance (MDR)-1 protein was strongly expressed in MEG-01. When cyclosporin-A, a modulator of MDR-1, was given with GO to MEG-01, GO-induced cytotoxicity was significantly increased, while no such effect was observed in HL-60. It suggests that p-glycoprotein-mediated efflux of GO is one of the resistance mechanisms in MEG-01. Disclosures: No relevant conflicts of interest to declare.

Abstract 2284 Poster Board II-261 Background: To evaluate the role of autologous and allogeneic SCT in the treatment of PTCLs, Japan Study Group for Cell Therapy and Transplantation conducted a multicenter retrospective survey in Japan and Korea. Methods: After excluding patients with adult T-cell leukemia/lymphoma and NK-cell tumors, patient data were newly collected from 330 patients (222 male and 108 female) with a median age of 49 years (range, 13–71) who underwent SCT between 9/1991 and 12/2008 (196 autologous and 134 allogeneic including 31 patients with previous autograft). Allogeneic SCT (53 BM, 54 PB, 1 BM+PB, 26 CB) was performed using a reduced-intensity conditioning (RIC) in 84 patients (63%). While a pathologic central review will be performed, currently there were 159 (48%) patients with PTCL, not otherwise specified, 63 (19%) with angioimmunoblastic T-cell lymphoma, 47(14%) with anaplastic large cell lymphoma (23 ALK-negative, 14 ALK-positive and 10 unknown), 12 (4%) with enteropathy-associated T-cell lymphoma, and others. The disease status at transplant in the allo-group was significantly worse than that in the auto-group (Table 1). The median number of chemotherapy regimens was 2 (range, 1–7), and the median duration between diagnosis and transplant was 267 days (range, 120-4889 days). Results: The median follow-up for surviving patients was 45 mo (range, 2.3–141 mo). There was no significant difference in overall survival among different groups, including histological subtypes, RIC and myeloablative conditioning in the allo-group and high-dose chemotherapy regimens in the auto-group. Early survival rate after transplant was significantly better for auto-group than allo-group (Wilcoxon P=0.001), but the difference was marginal in the total course (Logrank P=0.06) (Figure). The non-relapse mortality (NRM) in the auto-group was significantly lower than that in the allo-group (P1), cell source (CB/BM+PB), and performance status (PS; >1), stage, chemorefractory disease, international prognostic index (IPI; H-I/H risk) and prognostic index for PTCL, unspecified (PIT; group 3/4) at transplant. The risk factors in the allo-group were bulky mass at diagnosis, age (>50 years), cell source, and PS, stage, IPI and PIT at transplant, while those in the auto-group were age (>40 years), recurrence after frontline therapy, number of chemotherapy regimens, and stage, chemorefractory disease, IPI and PIT at transplant. Conclusions: Despite a worse disease status at transplant in the allo-group, the overall survival was comparable to that in the auto-group. This supports the notion that early allogeneic SCT is a valuable treatment option for PTCLs, although a large-scale randomized trial to identify a suitable upfront-transplant type for chemosensitive patients with PTCLs is warranted. Disclosures: No relevant conflicts of interest to declare.

Background: Pirarubicin (THP) is an anthracycline which was introduced to the Japanese market due to less cardiotoxicity compared to doxorubicin. The efficacy of THP-COP regimen (THP, cyclophosphamide (CPM), vincristine (VCR) and prednisolone (PSL)) was compared with CHOP in elderly patients with non- Hodgkin’s lymphoma in the previous Japanese randomized trial [Int J Hematol 81, 246–254, 2005]. The subset analysis showed peripheral T cell lymphoma (PTCL) had significantly better complete response (CR) rate with THP-COP than that of CHOP, whereas no such difference was observed in patients with B cell lymphoma. Methods: We underwent a multicenter phase II study to investigate the efficacy of THP-COP as a first line treatment for PTCL and adult T cell leukemia/lymphoma (ATLL). THP 50 mg/m2, CPM 750 mg/m2, and VCR 1.4 mg/m2 were given intravenously on day 1. PSL 60 mg/m2 was administered orally on days 1–5. The treatment was repeated at a 21-day interval, and its efficacy and toxicity were investigated. Results: Fifty-three patients aged from 38 to 83 (median 66) were entered into this study. Twenty-six patients were male and 27 female. Seventeen patients had PTCL including PTCL-U and AILT, and 36 ATLL. Stage IV disease was seen in 33, stage III in 15, and stage II in 5 patients. Thirty-four (64%) patients responded to THP-COP, including 17 (32%) CR and 17 (32%) partial response (PR). The median time to progression and overall survival were 9.5 and 17.5 months, respectively. Grade 3 to 4 neutropenia, anemia and thrombocytopenia occurred in 38 (72%), 18 (34%) and 31 (58%) patients, respectively. Twenty-seven (51%) patients developed febrile neutropenia, while grade 3 of nausea, anorexia, fatigue, arrhythmia, cardiac ischemia, sensory neuropathy, hypoalbuminemia, hyperbilirubinemia, and elevated serum AST/ALT were observed in 1 to 6 (2–11%) patients. No other adverse events greater than grade 3 were encountered. Conclusion: THP-COP is an effective and well-tolerated treatment regimen in patients with PTCL and ATLL as a first-line therapy.

Gemtuzumab ozogamicin (GO) is a humanized anti-CD33 antibody conjugated to a derivative of a tumoricidal antibiotic calicheamicin. GO alone induces complete remission in 26% of AML patients in first relapse. One way to improve the response rate is to combine GO with other agents. A quantitative relationship has been shown between rates and degrees of CD33 expression and GO-induced cytotoxicity in vitro by using gene transfer to manipulate CD33 expression in myeloid cell lines. We first studied CD33 expression on myeloid leukemia cells by stimulating with several agents in vitro, and found that ubenimex, but not G-CSF, M-CSF, or ATRA, increased CD33 expression on both HL-60 and K562 cells. Ubenimex is an aminopeptidase inhibitor isolated from Streptomyces olivoreticuli and commercially available as an oral agent for clinical use for AML treatment. To investigate whether ubenimex enhances the cytotoxicity of GO, HL-60 and K562 cells were cultured with GO and/or ubenimex for 3 days. When GO was administered, cell viabilities of HL-60 and K562 were reduced to 31.5% and 90.3% as compared with control, respectively. Treatment with ubenimex alone did not influence viabilities of either HL-60 or K562. However, when cells were preincubated with ubenimex and then cultured with GO, cell viabilities decreased to 18.5% and 81.4% for HL-60 and K562, respectively, indicating that pretreatment with ubenimex enhanced GO-induced myeloid leukemia cell death in vitro. We next assessed the mechanism of cell death. The treatment with GO alone and ubenimex alone induced apoptosis in 39.2% and 2.9% of HL-60 cells, respectively. When HL-60 cells were preincubated with ubenimex and then cultured with GO, the number of apoptotic cells increased to 62.9%, demonstrating that ubenimex augments GO-induced apoptosis. Our data suggest that the priming of AML cells with ubenimex should improve the clinical efficacy of GO.

Chemokine (C-X-C motif) receptor 4 (CXCR4) plays an important role on hematogenous metastasis of tumor cells to the organs comprising constitutively stromal cell-derived facter-1 (SDF-1). Retinoids have been utilized as a drug for treatment of acute promyelocytic leukemia (APL), in which tumor cells give rise to differentiation and cell death by activation of retinoic acid receptors (RARs). However, the effects of retinoid on chemokine-mediated cell motility are totally unknown. In this study, we examined the effects of retinoids, including all-trans retinoic acid (ATRA) and RARα-specific retinoid Am-80, on CXCR4-mediated chemotactic activity using five myeloid and lymphoid leukemia cells (HL-60, K562, MOLT-3, Jurkat, and U266B1). When cells were incubated with ATRA or Am-80 at doses up to 10 mM for 3 days, comparable effects were found; cytotoxic effect occurred only in HL-60, and growth inhibition was albeit detected in all of other cells. The extents of cell surface CXCR4 in all of the cells detected by flow cytometric analysis tended to be correlated with SDF-1-induced migration activity. Cell surface CXCR4 was slightly decreased by ATRA treatment, while it was significantly decreased by Am-80 treatment, and the alterations were in time- and dose-dependent fashions. In terms of CXCR4 expression, Jurkat was the most sensitive cells. Cell migration in response to SDF-1, was monitored by a modified Boyden chamber method. SDF-1 induced significant chemotaxis in HL-60, MOLT-3 and Jurkat cells, but did not in K562 and U266B1 cells. After treatment with the retinoids, HL-60 was only a cell type increasing random cell motility, however in retinoids-pretreated MOLT-3 and Jurkat cells, no such effect was found, and therefore suggesting that up-regulation of the motility in HL-60 is maybe due to a response by dyeing cells. However, in a case of pretreatment with not ATRA but Am-80, SDF-1-induced chemotaxis of MOLT-3 and Jurkat cells was dose-dependently repressed in accordance with decrease in cell surface CXCR4 expression. This was also confirmed by a time-laps horizontal migration system, indicated that pseudopodia formation accompanied with F-actin rearrangement by SDF-1 was strongly inhibited by Am-80-pretreatment. These results indicate that RARα-specific retinoid Am-80 may induce not only growth inhibition, but also prevention of SDF-1/CXCR4-mediated hematogenous metastasis in CXCR4 expressing tumor cells.

De novo CD5-positive diffuse large B-cell lymphoma (CD5+ DLBCL) is well known to have clinicopathologically and genetically different characteristics from CD5-negative (CD5−) DLBCL and mantle cell lymphoma. To obtain more clinicopathologic information of CD5+ DLBCL, we reviewed specimens from 128 cases of CD5+ DLBCL and analyzed the relationship between the morphologic features and a long-term survival. The current series includes the 109 CD5+ DLBCL cases described in our previous study (Yamaguchi M, Blood 2002). All patients were diagnosed between 1984 and 2002 as having DLBCL according to the WHO classification. They had no past history of any other lymphoproliferative disorders and were immunohistochemically confirmed to be cyclin D1-negative. Four morphologic variants were identified: common (monomorphic) (n=97), pleomorphic (n=15), giant-cell rich (n=14) and immunoblastic (n=2). Intravascular or intrasinusoidal infiltration was seen in 41% of the cases, and bi-nucleated snowman-like cells were seen in 80% of the cases. BCL2 protein expression in CD5+ DLBCL was more frequent than that in CD5− DLBCL (P=0.0009). Immunohistochemical analysis in a serial 45 cases of CD5+ DLBCL revealed that 80% of CD5+ DLBCL cases (36/45) were classified as non-germinal center B-cell type DLBCL. CD10 was positive in 8 cases (18%), and MUM1 was positive in 93% of the cases (42/45). The estimated 5-year overall survival rate was 36% after a median observation time of 79 months. Patients with the common variant showed better prognosis than those with the other three morphologic variants (P=0.018). Multivariate analysis showed that age more than 60 at diagnosis [hazard ratio (HR) 2.25, 95% confidence interval (CI) 1.36–3.73], advanced stage (HR 1.76, 95%CI 1.06–2.93), high serum LDH level (HR 2.2, 95%CI 1.21–4.00), and histopathologic categorization (not common) (HR 1.6, 95%CI 1.00–2.59) were independent prognostic factors for survival. In all, 16 patients (13%) developed central nervous system (CNS) recurrence. Our study revealed the morphologic spectrum and a high incidence of CNS recurrence in CD5+ DLBCL, and confirmed that CD5+ DLBCL shows frequent BCL2 protein expression and it is mainly regarded as non-germinal center B-cell type of DLBCL.

The chemokine stromal cells derived factor-1 (SDF-1)/CXCL12 stimulates lymphocytes, hematopoietic stem cells (HSC) and hematopoietic progenitor cells (HPC) through the corresponding chemokine receptor CXCR4, which modulates homing, retention, and engraftment into the bone marrow. However, regulatory factors for CXCR4 excluding cytokines are totally unknown. Besides, all-trans retinoic acid (ATRA), a drug inducing cell differentiation and apoptosis, increases motility with unknown mechanisms. In the present study, we investigated the intracellular regulatory mechanisms of SDF-1α-induced migration of ATRA-treated cells. We initially surveyed cells exhibiting strongly membrane-associated CXCR4 (M-CXCR4), and found 3 out of 10 different cell lines (MOLT3, U266B1, and Raji). Of those, MOLT3 was identified as the only one enabled to respond to SDF-1α chemoattraction, accompanied by an increase in actin polymerization. When M-CXCR4 in MOLT3 was neutralized or antagonized by an antibody or the antagonist AMD3100, the migration activity to SDF-1α, analyzed using the Boyden chamber method, was totally abolished. ATRA pretreatment for 3 days induced growth inhibition of MOLT3, but apoptosis and differentiation did not occur. ATRA treatment alone had no effects on either CXCR4 gene expression or its total protein expression, and cell motility also did not increase; however, ATRA treatment dose dependently increased M-CXCR4 expression. When SDF-1α was added to the lower chamber, migration activity of cells pretreated by different doses of ATRA increased dose dependently and reached up to 5 times greater compared with cells lacking ATRA pretreatment. The increased migration activity was blocked by wortmannin, which inhibits phosphatidylinositol 3 kinase (PI3K), AKT inhibitor, and cytochalasin D, which inhibits actin polymerization; U0126, an inhibitor of MEK1/2, did not block the effects of ATRA and SDF-1α. The results indicate that M-CXCR4 is quite important for cell migration, and the intracellular signaling for the M-CXCR4/SDF-1 axis primarily involves the PI3K/Akt pathway, which ATRA positively affected. This is the fist time to show that migration activity to SDF-1 was accelerated by ATRA. Clinical application of this phenomenon may help to augment HSC/HPC homing after transplantation.

To assess the clinicopathological applicability and reproducibility of the new WHO classification against peripheral T/NK-cell lymphomas, International T-cell Lymphoma Study Group was organized. Our task is to show the clinicopathologic and therapeutic results of ATLL from International T-cell Lymphoma Project. In the study, 1159 cases of T/NK-cell lymphomas, excluding mycosis fungoides and Sezary syndrome, were collected from the 21 centers in 13 countries of North America, Europe and Asia. All cases were reviewed by expert hematopathologists. ATLL was diagnosed in 126 (10.9%) patients. It was rare in North America (2.0%), and Europe (1.5%), but frequent in Asia (24.8%). There were clinical subtypes of ATLL from smoldering, chronic, lymphoma to acute type, and 87% of the patients fell into lymphoma type. The median age was 60 (range; 15–92) and male to female ratio was 1. B symptoms, PS 2–4(ECOG) and two or more extranodal sites were found in 39 (31%), 29 (23%) and 43 (34%) patients, respectively. Bone marrow involvement was present in 28% of the patients. Stage I/II was seen in 10% of the patients, stage III in 18% and stage IV in 73%. There were 23 (19%) patients in score 0/1, 41 (33%) in score 2, 40(32%) in score 3 and 20(16%) in score 4/5 of International Prognostic Index (IPI). There were 20 (17%) patients with less than 150x103/ml of platelet counts, 107 (85%) with over 20% transformed T cells in absolute lymphocyte counts, and 113 (94%) with over 20% Ki-67 positive cells. The 5- year-overall survival (OS) and failure free survival (FFS) were 14.4% and 12.1%, respectively. The significant prognostic factors for OS were B symptoms (p=0.018), PS (p=0.0022), stage (p=0.019), extranodal sites (p=0.045) and IPI (p=0.019). By using the multivariate analysis, the important prognostic factors were found to be B symptoms, platelet counts (20%), and Ki67+ cells (>20%). However, the Cox model analysis showed that no significant prognostic factors were found after adjusting by IPI. For treatment, the combination chemotherapy using anthracycline as an initial therapy was given in 109 of 120 (91%) treated patients. The overall and complete response rates were 69% and 33%, respectively. It is unfortunate that 71 (63%) patients had relapsed. Many drugs were used including prednisone 105 (88%), vincristine 102 (85%), cyclophosphamide 92 (77%), doxorubicin 77 (64%), etoposide 53 (44%), mitoxantrone 14 (12%), cisplatin 11 (9%), methotrexate 8 (7%) , bleomycin 6 (5%) and cytarabine 5 (4%). There was no difference in OS and FFS according to the treatment arms includ.