ALBERT

Description: Abstract 3370 Poster Board III-258 Although the outcome of patients (pts) with adult T-cell leukemia-lymphoma (ATL) remains poor when they are treated with conventional chemotherapy, we previously showed in a multi-center prospective study that one-third of pts who underwent RIST from a related donor in CR or PR could survive without disease for more than 2 years (Tanosaki R et al., BBMT 2008). In this retrospective study, we reviewed our single-center experience with RIST for ATL pts, focusing on the outcome of those who underwent RIST in non-remission status or who relapsed after RIST. A total of 24 pts underwent RIST from a related donor between 2001 and 2008. The median age was 54 years (range, 44-65). Of the 14 males and 10 females, 19 were acute type and 5 were lymphoma type. Disease status at transplantation was 5 CR, 10 PR, 8 NC and 1 PD. Donors were siblings in 18 and children in 6, including 5 HTLV-1 healthy carriers. HLA in serology was 6/6 in 19 and 5/6 in 5. Stem cell sources were PBSC in 22 and BM in 2. Conditioning regimens were fludarabine (30 mg/m2 iv days -8 to -3) and busulfan (3.2 mg/kg iv days -6 and -5) with (n=8) or without (n=16) rabbit anti-T-lymphocyte globulin (ATG, Fresenius; 2.5 mg/kg iv days -2 and -1). All patients received cyclosporine alone for GVHD prophylaxis. Engraftment was rapid in all 24 pts (neutrophil〉500/uL; median 12 days, range 10 -19), with no graft failure. There were 3 non-relapse mortalities; respiratory failure from bronchiolitis obliterans at 21 months (mos), interstitial pneumonitis at 47 mos, and pneumococcal sepsis at 38 mos. Notably, 10 of the 19 pts who were non-CR at RIST survived without disease progression to a median of 53 mos (range, 20 to 85). All of these pts were acute type, and had circulating ATL cells in the peripheral blood (PB) immediately before RIST (average 33% of WBC, range 5-73). Circulating ATL cells decreased to below 5% within 1 mo in 8 pts. A total of 12 pts relapsed within 16 mos; 7 (58%) within 3 mos, and 11 (92%) within 12 mos. Two patients who had relapsed after RIST showed a significant but transient response to the withdrawal of immunosuppression (CR 1, PR 1). Donor lymphocyte infusion was performed in 6 pts without significant benefits. Seven pts who relapsed at a single site, which was confirmed by CT scan or FDG-PET, were treated with local irradiation alone, and 3 whose HTLV-1 proviral load in PB had become negative at relapse survived to 48, 64 and 77 mos; 1 pt required 2 courses of irradiation because of immediate relapse at the margin of the preceding radiation field, and another pt underwent surgical resection of a residual mass since a biopsy revealed a viable lesion at the irradiated site. The 5-year overall and progression-free survival of all pts were 52% (95% CI, 38-66%) and 37% (95% CI, 22-52%), respectively, at a median follow-up of 59 mos (range, 12 to 85) in surviving pts. HTLV-1 proviral load in PB was examined using real-time PCR for tax in 208 samples from 21 evaluable pts, and it became negative at least once in 15 pts (71%), including 1 pt whose donor was an HTVL-1 carrier; proviral load remained negative in 7 pts at a median follow-up of 32 mos (range, 3 to 84). Since HTLV-1 tax is a promising target molecule for identifying the immunological mechanism, HLA-restricted tax-specific CTLs were examined in HLA-A2- and/or A24-positive pts using tax tetramers by taking blood samples periodically after informed consent was obtained from each pt. A total of 80 samples in 13 pts were analyzed. The number of tax tetramer-positive (tax+) cells did not change significantly up to at least 1 year after RIST, while the clinical responses and decrease/disappearance of HTLV-1 proviral load were observed within 3 mos in most cases. An increase in tax+ cells was observed after 1 year in 2 pts who had achieved CR. In conclusion, about half of the acute-type ATL pts with a significant involvement of ATL cells in PB at RIST could survive for a long time in our cohort. ATL pts who relapsed at a single site after RIST still have a chance to be cured with local treatment using irradiation alone or surgical resection with the aid of HTLV-1 proviral load as a marker for minimum residual disease. Since most ATL pts had already become resistant to chemotherapy and the intensity of conditioning was reduced, potent GV-ATL and GV-HTLV-1 effects might have played a key role in disease control. However, tax-specific CTL kinetics did not correlate with either clinical responses or the HTLV-1 proviral load, which suggested that other molecules may be immunologically targeted. Our results might contribute to the establishment of the cure-oriented treatment strategy for ATL pts. Disclosures: No relevant conflicts of interest to declare.

Description: Background Intensive efforts of genome sequencing studies during the past decade identified 〉100 driver genes recurrently mutated in one or more subtypes of myeloid neoplasms, which collectively account for the pathogenesis of 〉90% of the cases. However, approximately 10% of the cases have no alterations in known drivers and their pathogenesis is still unclear. A possible explanation might be the presence of alterations in non-coding regions that are not detected by conventional exome/panel sequencing; mutations and complex structural variations (SVs) affecting these regions have been shown to deregulate expression of relevant genes in a variety of solid cancers. Unfortunately, however, no large studies have ever been performed, in which a large cohort of myeloid malignancies were analyzed using whole genome sequencing (WGS) in an attempt to identify a full spectrum of non-coding alterations, even though its efficacy have been demonstrated in many solid cancers. In this study, we performed WGS in a large cohort of pan-myeloid cancers, in which both coding and non-coding lesions were comprehensively analyzed. Patients and methods A total of 338 cases of myeloid malignancies, including 212 with MDS, 70 with AML, 17 with MDS/MPN, 23 with t-AML/MDS, and 16 with MPN were analyzed with WGS, of which 173 were also analyzed by transcriptome sequencing. Tumor samples were obtained from patients' bone marrow (N=269) or peripheral blood (N=69), while normal controls were derived from buccal smear (N=263) or peripheral T cells (N=75). Sequencing of target panel of 86 genes were performed for all samples. Sequencing data were processed using in-house pipelines, which were optimized for detection of complex structural variations (SVs) and abnormalities in non-coding sequences. Results WGS identified a median of 586,612 single nucleotide variants (SNVs) and 124,863 short indels per genome. NMF-based decomposition of the variants disclosed three major mutational signatures, which were characterized by age-related C〉T transitions at CpG sites (Sig. A), C〉T transitions at CpT sites (Sig. B), and T〉C transitions at ApTpN context (Sig. C). Among these, Sig. C showed a prominent strand bias and corresponds to COSMIC signature 16, which has recently been implicated in alcohol drinking. Significant clustering of SNVs and short indels were interrogated across the genome divided into different window sizes (1Kbp, 10Kbp, 100Kbp) or confining the targets to coding exons and known regulatory regions, such as promoters, enhancers/super enhances, and DNase I hypersensitive sites. Recapitulating previous findings, SNVs in the coding exons were significantly enriched in known drivers, including TP53, TET2, ASXL1, DNMT3A, SF3B1, RUNX1, EZH2, and STAG2. We detected significant enrichment of SNVs in CpG islands, and promoters/enhancers. We also detected a total of 8,242 SVs with a median of 15 SVs/sample, which is more prevalent than expected from conventional karyotype analysis. Focal clusters of complex rearrangements compatible with chromothripsis were found in 8 cases, of which 7 carried biallelic TP53 alterations. NMF-based signature analysis of SVs revealed that large (〉1Mb) deletions, inversions, and tandem duplications and translocations are clustered together and were strongly associated with TP53 mutations, while smaller deletions and tandem duplications, but not inversions, constitute another cluster. As expected, FLT3-ITD (N=15) and MLL-PTD (N=12) were among the most frequent SVs. Unexpectedly, in addition to known SVs associated with t(8;21) (RUNX1-RUNX1T1) (N=6) and t(3;21) (RUNX1-MECOM) (n=1) as well as non-synonymous SNVs within the coding exons (N=30), we detected frequent non-coding alterations affecting RUNX1, including SVs (N=15) and SNVs around splicing acceptor sites (N=5), suggesting that RUNX1 was affected by multiple mechanism, where as many as 38% of RUNX1 lesions were explained by non-coding alterations. Other recurrent targets of non-coding lesions included ASXL1, NF1, and ETV6. Conclusions WGS was successfully used to reveal a comprehensive registry of genetic alterations in pan-myeloid cancers. Non-coding alterations affecting known driver genes were more common than expected, suggesting the importance of detecting non-coding abnormalities in diagnostic sequencing. Disclosures Nakagawa: Sumitomo Dainippon Pharma Co., Ltd.: Research Funding. Usuki:Mochida Pharmaceutical: Speakers Bureau; Astellas Pharma Inc.: Research Funding; Sanofi K.K.: Research Funding; GlaxoSmithKline K.K.: Research Funding; Otsuka Pharmaceutical Co., Ltd.: Research Funding; Kyowa Hakko Kirin Co., Ltd.: Research Funding; Daiichi Sankyo: Research Funding; Celgene Corporation: Research Funding, Speakers Bureau; SymBio Pharmaceuticals Limited.: Research Funding; Shire Japan: Research Funding; Janssen Pharmaceutical K.K: Research Funding; Boehringer-Ingelheim Japan: Research Funding; Sumitomo Dainippon Pharma: Research Funding, Speakers Bureau; Pfizer Japan: Research Funding, Speakers Bureau; Novartis: Speakers Bureau; Nippon Shinyaku: Speakers Bureau; Chugai Pharmaceutical: Speakers Bureau; Takeda Pharmaceutical: Speakers Bureau; Ono Pharmaceutical: Speakers Bureau; MSD K.K.: Speakers Bureau. Chiba:Bristol Myers Squibb, Astellas Pharma, Kyowa Hakko Kirin: Research Funding. Miyawaki:Otsuka Pharmaceutical Co., Ltd.: Consultancy; Novartis Pharma KK: Consultancy; Astellas Pharma Inc.: Consultancy.

Description: Introduction The International Prognostic Index (IPI) is used to predict survival in patients with diffuse large B-cell lymphoma (DLBCL). The IPI includes five factors: age, stage, LDH, performance status, and extranodal sites. However, studies suggest that certain blood markers not included in the IPI, such as CRP, albumin, absolute lymphocyte count (ALC), and platelet count, predict survival. Recently, an enhanced IPI (NCCN-IPI) was proposed and showed excellent prognostic performance. It uses the same factors as the IPI, but refines the impact of age and LDH and the definition of extranodal disease. Few studies have assessed the prognostic impact of blood markers other than the five factors of the IPI in the context of the NCCN-IPI. Here, we analyzed the prognostic role of simple blood markers in the NCCN-IPI. Patients and Methods Data from consecutive DLBCL patients diagnosed between January 2004 and June 2014 were retrospectively analyzed. Patients receiving rituximab and anthracycline-based chemotherapy were included in the analysis. Values considered positive were CRP 〉1 mg/dl, albumin

Description: Introduction: Follicular lymphoma (FL) is the second most common type of non-Hodgkin cell lymphoma, and usually manifests as a disseminated disease. Bone marrow (BM) involvement, which occurs in 40-70% of cases, is often seen in follicular lymphoma and thought to be associated with less favorable prognosis. Diagnosis of BM involvement has traditionally been based on morphological findings, and BM involvement has been determined using histology alone in most clinical trials. Immunocytologic or molecular studies, such as flow cytometry (FCM) and polymerase chain reaction (PCR), have become more readily available, and their usage has clearly documented minimal BM involvement reproducibly. In this study, we evaluated the impact of BM involvement detected by FCM and PCR on the outcome of patients treated for FL. Methods: Patients who were diagnosed with biopsy-proven FL between 2004 and 2015 at our institution were included in the study. All patients had received a staging bone marrow examination before treatment with immunotherapy-based regimen. Immunocytologic [FCM] and/or molecular [PCR] studies were always performed if the patients did not have morphological BM involvement. We used 4- or 6- color FCM, and performed PCR analysis of Bcl-2/IgH rearrangement and/or IgH rearrangement detected by modified BioMed-2 protocol. A total of 90 patients were included, and the median follow-up duration was 36 months (range, 6|122 months). The BM status was classified using into 3 categories: morphological, minimal, and negative BM involvement. Minimal BM involvement was defined as BM involvement detected by FCM or PCR without morphological evidence. Morphological and minimal BM involvements were detected in 37 (41%) and 38 (42%) patients, respectively. The primary outcome measure was progression-free survival (PFS). PFS curves were plotted using the Kaplan-Meier method and compared by the log-rank test. Multivariate analyses were performed using a Cox linear regression model. There were significant differences in gender, LDH levels, stage, nodal sites, and FL International Prognostic Index (FLIPI) between patients with and without morphological BM involvement (Table1). Results: The 3-year PFS rate for patients with negative BM involvement was significantly better than that for patients with minimal or morphological BM involvement (84.8% vs. 40.3% vs. 60.5%; p= 0.043) (Figure 1). There was no statistical difference in 3-year PFS between patients with morphological BM involvement and those with minimal BM involvement. The difference of 3-year PFS rate between patients with minimal BM involvement and those with negative BM involvement was significant for patients with FLIPI low-intermediate risk (88.9% vs. 51.5%; p= 0.032) and those with advanced stage disease (90.0% vs. 33.6%; p= 0.027), but there were no significant differences in patients deemed FLIPI high risk and those with limited stage disease. Multivariate analysis revealed that BM involvement, including morphological and minimal involvement, was a significant poor prognostic factor (hazard ratio 4.885 [95% confidence interval 1.16-20.56], p = 0.0305). Conclusion: At the start of treatment, bone marrow involvement was seen in most FL patients. Patients without any BM involvement had an excellent prognosis. Patients with minimal BM involvement had an equally poor prognosis as those with morphologic BM involvement. Table 1 FLIPI: Follicular Lymphoma International Prognostic Index Table 1. FLIPI: Follicular Lymphoma International Prognostic Index Table 2 BM state positive: including morphological and minimal bone marrow involvement. Table 2. BM state positive: including morphological and minimal bone marrow involvement. Figure Figure. Disclosures Ishikawa: Mundipharma KK: Research Funding.

Description: Introduction: Primary graft failure or delayed engraftment is a well-studied complication of allogeneic hematopoietic stem cell transplantation (HSCT). According to previous studies, the following factors are associated with primary graft failure or delayed engraftment: underlying disease, disease status, human leukocyte antigen (HLA) disparity between recipient and donor, type of conditioning regimens, stem cell source, and stem cell dose. As engraftment failure or delayed engraftment frequently brings uncontrollable infectious events for allogeneic HSCT recipients, the prevention of primary engraftment failure and the promotion of early engraftment is critically required. Splenomegaly is associated with primary engraftment failure and poor overall survival (OS) after allogeneic HSCT in patients with myelofibrosis. As patients with acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) sometimes develop splenomegaly, we investigated the effect of splenomegaly on engraftment kinetics in patients with AML and MDS. Patients and methods: A retrospective case analysis was performed for 87 patients with AML or MDS who were treated with allogeneic HSCT at our hospitals from April 2010 to March 2016. Two patients without computed tomography data just before allogeneic HSCT were excluded. The remaining 85 patients were included in the analysis. Spleen volume was evaluated using iPlan. We set the primary endpoint as neutrophil engraftment (〉500/mL) at 28 days after allogeneic HSCT. We also evaluated OS, progression-free survival (PFS), cumulative incidence (CI) of non-relapse mortality (NRM) and CI of relapse (CIR). To provide simple risk stratification, we also employed survival classification and regression tree (CART) analysis, and divided study patients into two risk groups. To evaluate the impact of spleen volume on the endpoint, we employed multivariable Cox regression analysis for OS and PFS and Fine and Gray analysis for engraftment; CI of NRM and CIR were calculated using spleen volume as the continuous variable. Adjusted covariates were as follows: age, sex, disease, disease status, stem cell source, and HLA mismatch. Results: Median spleen volume was 145 cm3 (38-1661 cm3) among the study patients. The survival CART analysis revealed that the discriminating spleen volume to discern engraftment was 320 cm3. Thus, we defined patients with a spleen volume above 320 cm3 as part of the splenomegaly group, and the remaining patients as part of the non-splenomegaly group. The neutrophil engraftment rate at 28 days after allogeneic HSCT was 37.5% and 93.5% (p = 0.021) in the splenomegaly and non-splenomegaly groups, respectively (Figure 1). OS at 2 years was 37.5% and 72.6% (p = 0.011), PFS at 2 years was 30% and 67% (p = 0.005), CI of NRM at 1 year was 27.5% and 5.6% (p = 0.044), and CIR at 1 year was 42.5% and 26.5% (p = 0.34) in the splenomegaly non-splenomegaly groups, respectively (Figure 2). To further assess the influence of spleen volume on engraftment and other outcomes previously described, we performed multivariate Cox regression analysis or Fine and Gray analysis. The adjusted HR (95% CI) for spleen volume per 100 cm3 was 0.79 (0.68-0.92, p = 0.003) on engraftment, 1.23 (1.04-1.47, p = 0.017) on OS, 1.27 (1.08-1.5, p = 0.005) on PFS, 1.64 (1.09-2.4, p = 0.016) on CI of NRM, and 0.96 (0.83-1.1, p = 0.59) on CIR. Conclusion: Our study showed that spleen volume predicts poor engraftment rate and poor PFS due to increased NRM. Patients with splenomegaly were at a higher risk of engraftment failure or delayed engraftment and may be considered for spleen irradiation before allogeneic HSCT. Disclosure of Interest: We declare no conflicts of interest. Disclosures Ishikawa: Mundipharma KK: Research Funding.

Description: Abstract 4542 Background: Norovirus-gastroenteritis (NV-GE) is considered as a highly transmittable disease that can lead to fatal outcomes in vulnerable populations. Therefore, prompt detection of norovirus in stool specimens is important for patients who undergo hematopoietic stem cell transplantation (HSCT). The commercially available immunochromatography kit (Denka Seiken, Tokyo, Japan) is a diagnostic tool that can easily and rapidly detect norovirus antigens with high specificity and relatively less sensitivity compared with reverse transcription polymerase chain reaction (RT-PCR). Although previous studies used RT-PCR to detect norovirus in stool, this method is not necessarily a standardized technique for the clinical use, and only limited information is available about clinical significance of norovirus infection among the HSCT recipients. Here, we report an analysis of patients with NV-GE in our HSCT unit using the immunochromatography method. Patients and Methods: We prospectively examined stool specimens to detect norovirus antigens in patients who developed diarrhea in our HSCT unit between December 2008 and June 2011. We also retrospectively examined stool specimens which had been collected for Clostridium difficile (CD) toxin test and frozen at –40°C between January 2007 and November 2008. During the study period, a total of 468 patients underwent HSCT (autologous 83, allogeneic 385) at our center and we tested stool specimens from 355 patients who developed diarrhea. Results: Norovirus was detected by the immunochromatography method in 11 patients among the HSCT recipients, and in 1 patient who died before HSCT because of disease progression. Among the 12 patients with NV-GE, 3 were retrospectively diagnosed using the frozen specimens. The CD toxin test was also positive in 1 of the 12 patients with NV-GE. The median age of the 12 patients was 56 years (range, 29–66). The median duration of symptoms was 30 days (2–134). Among the 11 patients who developed NV-GE after HSCT, primary disease included lymphoma (6 patients), acute leukemia (4 patients), and multiple myeloma (1 patient), and 9 of them were not in remission at HSCT. One patient developed NV-GE after autologous HSCT, and 10 patients after allogeneic HSCT. Among the 10 allo-HSCT recipients, 6 received grafts from unrelated bone marrow donor and 4 received grafts from related donor. Five patients received myeloablative conditioning and 5 received reduced-intensity conditioning before allo-HSCT. The median time from HSCT to the onset of symptoms was 36 days (4–93). The median time from HSCT to diagnosis of NV-GE was 37 days (11–101). At diagnosis of NV-GE, all allo-HSCT recipients were given immunosuppressive agents, and 2 of them received corticosteroids for intestinal graft-versus host disease (GVHD). The volume of diarrhea was more than 500 ml per day in 4 patients. Among 4 patients who underwent endoscopy of the lower gastrointestinal tract, intestinal GVHD was diagnosed in 2 patients by histopathology findings, whereas the other 2 patients had no evidence of intestinal GVHD, which resulted in no need for an intensification of immunosuppression. Of the 12 patients with NV-GE, 6 were alive with a median follow-up of 826 days (17–1168) after the diagnosis of NV-GE. No patients died of NV-GE, and 6 died of other causes (disease progression, 4; GVHD, 1; multiple organ failure, 1). There was no outbreak of NV-GE as we promptly implemented isolation of infected patients and enhanced hygiene strategy within an hour of collection of stool specimens in patients with diarrhea. Conclusions: In this study, we detected 11 patients who developed NV-GE after HSCT using the immunochromatography method. Our results suggested that this method is helpful in the differential diagnosis of patients with diarrhea after HSCT and enable us to take an appropriate and prompt preventive measure. In the future study, validation with RT-PCR and immunochromatography method is warranted among the immune-compromised populations. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 2318 Introduction: ES has been well recognized to increase non-relapse mortality (NRM) in autologous hematopoietic cell transplantation (HCT). However, little information is available on ES in allogeneic HCT, particularly after reduced-intensity conditioning. To determine the incidence, risk factors and outcome of ES after RIST, we retrospectively reviewed the medical records of 285 patients (pts) who underwent RIST at our institution between 1999 and 2007. Patients and Methods: After excluding 37 pts who received cord blood transplantation or a second or subsequent allogeneic HCT, and those who experienced primary graft failure, 248 pts (male 153, female 95) with a median age of 55 y (range, 21–68) fulfilled the criteria of this study. The underlying diagnosis included lymphoma (106 pts; 43%), acute myeloid leukemia (59; 24%), myelodysplastic syndrome (51; 21%), and others (32; 13%). Ninety-two pts (37%) had a standard-risk disease such as acute leukemia or lymphoma in CR1, and 156 (63%) had a high-risk disease. The donor and stem cell source were related PBSC in 170 pts (69%), related BM in 8 (3%), and unrelated BM in 70 (28%). All of the pts received reduced-intensity conditioning with a purine analog and a busulfan-based regimen. With the use of G-CSF after HCT, neutrophil engraftment (ANC ≥500/μ l) was achieved at a median of 12 days (range, 5–28). ES was diagnosed when the patient showed at least 2 of the following symptoms within 96 h of neutrophil engraftment: (1) fever (〉38.0°C) without an identifiable source of infection, (2) skin rash (〉25% of body surface area) that was not due to a drug reaction, (3) weight gain (〉2.5% of baseline body weight), and (4) hypoxia (sPO2

Description: Abstract 270 BACKGROUNDS: A20 (also called TNFAIP3) gene encodes an ubiquitin-modifying enzyme involved in termination of NF-kB responses, and that is negative regulator of NF-kB. We previously reported that A20 is a common genetic target in B-cell lymphomas by using a genome–wide analysis of genetic lesions in 238 B-cell lymphomas. [Kato et al., Nature 459; 712, 2009] In the study, we showed that three Hodgkin's lymphoma (HL)-derived cell lines have deletion of both alleles or loss of one allele and a mutated allele of A20 gene by high-density typing using CNAG/AsCNAR algorism, which enabled accurate determination of allele-specific copy numbers, and thus allowed for sensitive detection of loss of heterozygosity (LOH) without apparent copy-number reduction, even in the presence of up to 70–80% normal cell contamination. Twenty-four primary samples from HL were also analyzed for the mutations and microdissected CD30-positive tumor cells (Hodgkin-Reed-Sternberg (HRS) cells) revealed one intronic and four missense mutations. However, we might have missed the homozygous deletions, which are shown in other B-cell lymphoma subtypes, and underestimates the frequency of involvement of A20 gene in primary HL cases. MATERIALS AND METHODS: The objects were 12 of the 24 samples of our previous cohort. The samples included one case that showed mutation in the previous study. We tried to detect A20 gene deletion in HRS cells with the combination of anti-CD30 immunofluorescence with FISH (fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasm: FICTION) method. BAC clones suitable for FISH analysis of A20 gene were screened. A BAC clone RP11-783B20 (TNFAIP3 locus, 6q23) labeled with spectrum orange by nick translation, was selected because of the best signal/noise ratio. CEP 6 Spectrum Green Probe was used to detect centromere of chromosome 6 (6p11.1-q11). Approximately 4 micrometer thick slides were immunostained with anti-CD30 antibody and FISH was performed. As the diameter of HRS cells is known to be several times more than the 4 micrometer thickness of the section slides suitable for FISH, ratio of A20/CEP 6 signals were calculated and the A20 gene status was evaluated in CD30 positive cells. A20 and CEP6 signals were also counted in CD30 negative cells and used as a control. We counted signals of 30(15–55) CD30 positive cells and 50 CD30 negative cells. RESULTS: In 9 cases among 12 cases, signals were successfully counted. The average number of CEP6 signals on the CD30 negative cells was 1.53, which showed that a whole cell is located on the section slides. The ratio of A20/CEP6 signals on these CD30 negative cells were approximately 1.0 confirming the quality of FISH signals. As predicted, the average number of the CEP6 signals on the HRS cell was approximately 1.21, which suggested that one HRS cell is cut in the middle and divided into pieces during preparation of the section slides. In these CD30 positive cells, the A20/CEP6 ratio was less than 1.0; the cells contained less A20 gene signals than CEP6 signals. The relative numbers of signals determined by FICTION in 9 cases are shown in the table. We knew that the case #3 had A 20 gene mutation determined by direct sequence analysis of micro-dissected CD30 positive cells. As the sequence analysis showed no residual wild A20 genes, the relative ratio of 0.71 was considered as a consequence of loss of normal allele. Thus, the cut-off level of 0.71 was set to define deletion of at least one allele. Two cases showed that the A20/CEP6 signal ratio was more than this figure (0.90 in case #1, and 0.80 in case #4). Except for these 2 cases, 7 out of 9 cases were judged to have deletions of A20 gene. Notably case #6 had the ratio of 0.14, which suggested that virtually complete loss of A20 gene had occurred. The sequence analysis had missed the deletions probably due to the contaminated surrounding cells. CONCLUSIONS: We found a frequent deletion of A20 gene in HL cases including a case with a complete loss. The involvement A20 gene deletion associated with the pathogenesis of HL might be more frequent than previously suggested by the sequence analysis. Disclosures: No relevant conflicts of interest to declare.

Description: Background: High-dose cytarabine (HD-AraC) is a promising therapy for acute myeloid leukemia (AML). However, it causes substantial toxicity, such as severe cytopenia and cytarabine-encephalopathy. The efficacy of HD-AraC for the treatment of core-binding-factor (CBF) AML has been well established; however, whether or not HD-AraC is suitable for the treatment of non-CBF AML is unclear. Moreover, it remains controversial whether HD-AraC is effective before allogeneic hematopoietic stem cell transplantation (allo-HSCT) for the treatment of non-CBF AML. In this study, we investigated the efficacy of HD-AraC in terms of optimal numbers of HD-AraC cycles required to treat non-CBF AML patients who did and did not receive allo-HSCT. Methods: We retrospectively collected the data of consecutive AML patients who were aged 15-70 years, treated in our hospital and diagnosed between Jan. 2000 and Apr. 2014. Patients with acute promyelocytic leukemia (APL) and CBF AML were excluded. Only patients who achieved complete remission (CR) and received post-remission therapies with a curative intent were included; namely, only those who received allo-HSCT or completed more than one cycle of consolidation chemotherapy. Patients who exclusively received azacytidine-based or low-dose AraC-based therapies were excluded. HD-AraC therapy was defined as a chemotherapeutic regimen utilizing 12 g/m2/cycle or more of AraC. Patients who were not suitable for HD-AraC because of comorbidities were also excluded. Overall survival (OS) was separately analyzed in patients who underwent allo-HSCT in the 1st CR (group A) and in those who did not (group B). The unadjusted probabilities of OS were estimated using the Kaplan-Meier method, and univariate analysis was done using the log-rank test. Multivariate analysis was performed with Cox proportional hazard regression models, and 95% confidence intervals (CIs) were calculated. In multivariate analysis, the number of cycles of HD-AraC, age, cytogenetic data, and whether or not the patient achieved CR after the 1st cycle of chemotherapy were used as covariates. In group A, donor source and conditioning were also considered as covariates. Results: A total of 166 non-APL, non-CBF AML patients were identified. Seventy-one patients were excluded for the following reasons: 50 did not achieve CR, 13 underwent incomplete consolidation therapies because of complications or early relapse, 7 had missing data, and 1 had chronic renal disease. For HD-AraC, toxicity was not a consideration for the discontinuation of treatment, but the dose of AraC was reduced to an intermediate dose (AraC2 cycles of HD-AraC were 50.6%, 85.7%, 75.0%, and 68.6% (n=22, 9, 8, 11) in group A, and 44.3%, 16.0%, 65.6%, and 50.0% (n=19, 10, 10, 6) in group B, respectively. Univariate analysis showed that patients in group A who received one or more cycles of HD-AraC had a longer 3y-OS (75.4% vs 50.6%, p=0.024) (figure 1), whereas patients in group B who received two or more cycles of HD-AraC had a longer 3y-OS (57.1% vs 36.2%, p=0.078) (figure 2). Multivariate analysis of patients in group A showed that one or more cycles of HD-AraC resulted in a survival benefit (Hazard ratio=0.26, 95%CI: 0.070-0.98, p=0.046), whereas multivariate analysis of patients in group B showed that two or more cycles of HD-AraC (Hazard ratio=0.36, 95%CI: 0.13-0.97, p=0.044) and intermediate-risk of cytogenetics resulted in a survival benefit. Conclusion: For consolidation chemotherapy, at least one cycle of HD-AraC is recommended for patients who plan to receive allo-HSCT, and at least two cycles is required for other patients. Figure 1 Figure 1. Figure 2 Figure 2. Disclosures No relevant conflicts of interest to declare.

Description: The prognostic impact of interim positron emission tomography scans (I-PET) for patients with diffuse large B-cell lymphoma (DLBCL) is a matter of debate because its positive predictive value (20–80%) is low. Here, we aimed to improve the prognostic impact of I-PET by combining it with interim analysis of serum soluble interleukin-2 receptor (sIL2R) levels, the levels of which at the time of diagnosis are associated with the prognosis of DLBCL. We retrospectively examined data from DLBCL patients diagnosed at our institution between January 2006 and October 2013. Patients were included in the analysis if they met all of the following criteria: six or more cycles of rituximab plus CHOP regimen (R-CHOP); I-PET performed after 2-4 cycles of chemotherapy; and sIL2R levels measured after each cycle. Patients with primary central nervous system lymphoma, primary mediastinal large B-cell lymphoma, or transformed DLBCL from indolent lymphoma were excluded. I-PET was assessed visually according to the International Harmonization Project criteria. The interim sIL2R (I-IL2) level was defined as the value measured just before the fourth R-CHOP cycle. I-IL2 levels 〉 800 U/ml, or 2000 U/ml if serum creatinine was 〉 2.0 mg/dl (sIL2R is influenced by renal function), were regarded as positive. The primary endpoint of the study was progression-free survival (PFS). The unadjusted probabilities of PFS were estimated using the Kaplan-Meier method. The log-lank test and multivariate Cox regression analysis were used to assess the prognostic values of each clinical variable. In total, 135 patients were enrolled. The median age was 66 years (range, 34–89) and 66 patients (48.9%) were male, 27 (20%) had an ECOG performance status 〉1, 18 (13.3%) had bulky disease, 81 (60%) had advanced disease, and 61 (45.2%) had a high or high-intermediate International Prognostic Index (IPI) score. The median follow-up time was 25.6 months (range, 6.3–88.7) and the 2 year progression-free survival rate (2-y PFS) of the entire cohort was 72.9% (95% confidence interval (CI), 63.8–80). I-PET and I-IL2 were positive in 47 (34.8%) and 15 (11.1%) patients, respectively. Univariate analysis revealed that a high IPI score, a positive I-PET, and a positive I-IL2 had statistically significant poor prognostic effects on PFS, although gender and bulky disease did not. The three significant variables were entered into multivariate analysis, which identified positive I-PET and I-IL2 values (but not IPI) as independently associated with a poor prognosis. The 2-y PFS was 81.8% (95% CI, 70.4–89.1) for I-PET-negative and 56.3% (95% CI, 40.7–69.3; p