bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitation of EBV-DNA In Peripheral Blood In Hodgkin Lymphoma: Associations with Other Biomarkers and Patient Characteristics
Authors: Hohaus Stefan , Giachelia Manuela , Vannata Barabara , Santangelo Rosaria , Martini Maurizio , Cenci Tonia , Massini Giuseppina , Cuccaro Annarosa , Francesco D'Alo , Voso Maria Teresa , Fadda Giovanni , Larocca Luigi , Leone Giuseppe 
Published: October 2019

Abstract
Abstract 2678
The Epstein-Barr virus (EBV) is present in the malignant Hodgkin/Reed-Sternberg (HRS) cells of 20–40% cases of Hodgkin lymphoma (HL) in western countries. Detection and quantitation of cell-free plasma EBV-DNA has been proposed as a biomarker in EBV-associated malignancies, including HL. We analyzed the presence of EBV in HRS cells, peripheral blood compartments, and correlated these data to EBV serology, other circulating biomarkers and clinical characteristics. In a single institution cohort of 93 HL patients, EBV-DNA was quantitated using real-time PCR for the EBNA region in whole blood (WB, n=69), plasma (PL, n=75) and peripheral blood mononuclear cells (MNC, n=74). Cell-free DNA levels were determined using a real-time PCR for the globin gene (n=84), cytokine concentrations were measured in 85 patients. EBV status of HRS cells was studied using in-situ hybridization with EBER. EBV-DNA was positive in 32% WB, 21% PL and 20% MNC at HL diagnosis, with concordant WB and PL results in 86% of cases. EBER was present in HRS cells in 32% of cases. EBV-DNA copy number over 1500/ml was predictive for the presence of EBER in HRS cells in 86% and 88% WB and PL, respectively, while EBV copy numbers <1500 had only 55% and 57% PPV for WB and PL. Older patients (>50 years) were frequently EBER-positive (66%), with higher EBV copy numbers (median 12800/ml), while younger patients (<50 years) were less frequent EBER-positive (23%), and copy numbers were lower (median 1200/ml) (p=0.01). EBER-positive HL cases frequently had EBNA-1 antibody titers < 100 U/ml when compared to EBER-negative cases (44% vs. 13%, p=0.01), and there was a trend for a negative correlation between EBV copy number and EBNA1 titers (p=0.07). In patients with EBER+ HL the plasma EBV-DNA copy number correlated to the level of circulating cell-free DNA (rho=0.65), IL-10 levels (rho=0.87), IL-6 levels (rho=0.74) and inflammation parameters, including CRP (rho=0.79) and ESR (rho=0.57) (all p<0.02), and were higher in stage IV disease (p=0.02). In conclusion, a high EBV-copy number in whole blood and/or plasma at diagnosis is a good indicator for EBV-association of HL, and EBV copy numbers strongly correlate to other biological parameters of disease activity. Moreover, the inverse correlation between EBV copy number and EBNA1 antibody titers may indicate a reduction in immune surveillance favoring the expansion of EBV.

Disclosures:
No relevant conflicts of interest to declare.
Evaluation of the Prognostic Value of CD45RO+ and FOXP3+ Cells of the Micro-Environment In Classical Hodgkin Lymphomas Using Tissue Micro Array
Authors: Benet Claire , Ferrant Emmanuelle , Petrella Tony , Degrolard Emilie , Bastie Jean-Noel M. , Bonnotte Bernard , Casasnovas Rene-Olivier , Martin Laurent 
Published: October 2019

Abstract
Abstract 2687
Classical Hodgkin lymphomas (cHL) are B lympho-proliferative disorders that can be cured in 80% of cases. The goal of current research is to identify, at diagnosis, the 20% patients, who are likely to relapse or be refractory to treatment. As cHL have an extensive micro-environment implicated in tumour growth, the prognostic value (disease-free survival) of 2 important subsets of lymphoid cells (CD45RO+ effector memory T cells and FOXP3+ regulatory T cells) of the micro-environment has been evaluated using Tissue Micro Array (TMA).
Patients (n=96) treated and followed for cHL between 1998 and 2005 were included in this study. Six TMA with 3 spots per case, were built (3 formalin and 3 Bouin fixative). Immunostaining with anti-CD45RO (Dako, UCHL1, 1/400) and anti-FOXP3 (Abcam, 236A/E7 1/100) antibodies was performed on the Benchmark Ultra (Ventana). After scanning of the slides, the staining was quantified automatically using a function (pixel count) of Scanscope software (Aperio). ROC curves were used in order to calculate the best threshold for each marker, depending on the number and intensity of pixels. Survival curves were built according to these thresholds using the Kaplan Meier method and compared with the log-rank test. Then, a Cox model was used to estimate the risk of occurrence of an event in univariate and multivariate analyses.
The demographic characteristics of patients and sub-types of HL were not different in the << formalin >> and << Bouin >> groups. Median follow-up of patients was 69 months (1-135). In univariate analysis, low quantities of CD45RO+ cells was associated to a 40% 5-years progression free survival (PFS) compared to a 87% 5-years PFS for patients with high number of CD45RO+ cells (p=0.003), and low number of FOXP3+ cells was related to a 56% 5-years PFS compared to a 86% 5-years PFS for patients with high number of FOXP3+ cells (p=0.026). In multivariate analysis, taking into account the International Prognostic Score (IPS) and the expression of CD15 by neoplastic cells, the association of CD45RO and FOXP3 was the only independent prognostic factor identified (p<0.02 et RR=5.75).
This preliminary study underlines the importance of certain subsets of cells from the microenvironment of HL in the control of tumour growth. These results are encouraging. When used in association with validated criteria, they could make it possible to identify patients at risk of relapse and/or drug resistance. Nevertheless, due to the small number of patients in our study, these findings need to be validated in a larger and/or independent cohort.

Disclosures:
No relevant conflicts of interest to declare.
MMP-2 and MMP-9 Secreted by Leukemic Cells Increase the Permeability of Blood-Brain Barrier by Disrupting Tight Junction Proteins In Central Nervous System Leukemia
Authors: Feng Sa-ran , Chen Zi-Xing , Cen Jian-nong , Shen Hong-jie , Yao Li , Wang Yuan-yuan , Qi Xiao-fei , Chen Su-ning 
Published: October 2019

Abstract
Abstract 2694
Recurrence of acute leukemia (AL) in the central nervous system (CNS) confers a poor prognosis. However, little is known about the the underlying mechanisms of leukemic cell infiltration into the CNS. The blood brain barrier (BBB) makes CNS become a refugee to leukemic cells and serves as a resource of cells that seed extraneural sites. Tight junction in brain microvessel endothelial cells (HMECs) constituted by tight junction proteins is an important structure of BBB. Except the well known role of degrading extracellular matrix in metastasis of cancer cells, here we identify matrix metalloproteinase 2 (MMP-2) and MMP-9 secreted by leukemic cells are also associated with BBB opening by degrading tight junction proteins. We have successfully established an animal model of CNS leukemia by using a highly invasive human acute monocytic leukemia cell line SHI-1 in nude mice. Multiple organs including skull and brain were sectioned and determined histopathologically for leukemia cell infiltration. The fact that the down- regulation of ZO-1, Claun-5 and Occludin accompanied with up-regulation of MMP-2 and MMP-9 was correlated with BBB breakdown in mice with CNS leukemia was found when examined by laser scanning fluorescence confocal microscopy and gelatin in situ zymography. The treatment with MMP-inhibitor GM6001 could significantly reverse these changes in tight junction proteins. In an in vitro monolayer BBB model made by human encephalo-microvessel endothelial cells and matreil gel, MMP-2 and MMP-9 specifically down-regulated ZO-1, Claun-5 and Occludin. Knock-down or inhibition of MMP-2 and MMP-9 expression protected ZO-1, Claun-5 and occludin from degradation and alleviated the permeability of BBB. Our findings suggest that the degradation of tight junction proteins ZO-1, Claun-5 and Occludin by MMP-2 and MMP-9 secreted by leukemic cells constitutes an important mechanism in BBB breakdown in CNS leukemia. These studies provide a potent evidence for future pharmacological treatment to inhibit the CNS involvement in acute leukemia.

Disclosures:
No relevant conflicts of interest to declare.
Germline Genetic Variation and Treatment Response on CCG-1891
Authors: Sepe Dana , McWilliams Thomas , Chen Jinbo , Kershenbaum Aaron , Zhao Huaqing , La Mei , Davidas Meenakshi , Lange Beverly , Rebbeck Timothy R. , Aplenc Richard 
Published: October 2019

Abstract
Abstract 2711

Purpose:
Recent studies have suggested that polymorphisms in genes that encode enzymes involved in drug detoxification and metabolism may influence disease outcome in pediatric patients with acute lymphoblastic leukemia (ALL). We sought to extend current knowledge by using standard and novel statistical methodology to examine how frequency of polymorphism variants of certain drug metabolizing genes and gene pairs relate to relapse risk, toxicity, and drug dose delivery in standard risk ALL.

Methods:
We genotyped and abstracted chemotherapy drug dose data from treatment roadmaps on 557 patients on the Children's Cancer Group ALL study, CCG-1891. These data were analyzed with outcome data obtained from CCG. Fourteen common polymorphisms in genes involved in folate metabolism and/or phase I and II drug detoxification were evaluated individually by univariate analysis and clique-finding methodology was employed for detection of significant gene-gene interactions. Additionally, positive predictive value was calculated for those polymorphisms found to be statistically significant on univariate analysis.

Results:
After controlling for known risk factors, polymorphisms in four genes: GSTP1*B (HR=1.94, p=0.047), MTHFR (HR=1.61, p=0.034), MTRR (HR=1.95, p=0.01), and TS (3R/4R, HR=3.69, p=0.007), were found to significantly increase relapse risk. However, the positive predictive value of these genotypes was not statistically significant. One gene-gene pair, MTRR A/G and GSTM1 null genotype, significantly increased the risk of relapse after correction for multiple comparisons (p=0.012). Multiple polymorphisms were associated with various toxicities and there was no significant difference in dose of chemotherapy delivered by genotypes.

Conclusion:
These data suggest that various polymorphisms play a role in relapse risk and toxicity during childhood ALL therapy and that genotype does not play a role in adjustment of drug dose administered. Additionally, gene-gene interactions may increase the risk of relapse in childhood ALL and the clique method may have utility in further exploring these interactions.

Disclosures:
No relevant conflicts of interest to declare.
A Prognostic Score of Overall Survival In Adults with Acute Myeloid Leukemia
Authors: Al-Khabori Murtadha K. , Guyatt Gordon , Minden Mark D. , Yee Karen , Gupta Vikas , Schimmer Aaron , Schuh Andre , Brandwein Joseph 
Published: October 2019

Abstract
Abstract 2730

Background:
Acute myeloid leukemia (AML) is a malignant myeloid disorder with heterogeneous outcomes. A number of factors have been shown to be prognostic; age, white blood cell (WBC), prior malignancy, performance status (Eastern Cooperative Oncology Group; ECOG) and cytogenetics.

Methods:
We planned to develop and validate a prognostic score for the 5-year (y) overall survival (OS) of adults with AML receiving intensive induction chemotherapy. We used Cox model to estimate the regression coefficients and Kaplan-Meier to estimate the 5-y OS. We used Cox-Snell, Schoenfeld and deviance residuals for model diagnostics and bootstrap validation to estimate the performance measures; Harrell's concordance and deviance residuals.

Results:
We retrospectively analyzed 779 patients treated between 1998–2008, using a prospectively collected database. The median age was 58 y. Most patients had intermediate risk cytogenetics (61%) and good performance status (ECOG 0–1: 79%). The median follow up for the surviving patients was 26.7 months (95% CI 18.8–32.9 months). The 5-y OS was 26% (22- 30%). All variables were statistically significant in the multivariable Cox regression model; age (y) (Hazard Ratio, HR 1.02; 95% CI 1.018–1.034), WBC (1*10^9/L) (HR 1.004; 1.002–1.006), prior malignancy (HR 1.58; 1.26–2.00), ECOG (ECOG 2 HR 1.41; 1.06–1.88, ECOG 3–4 HR 9.99; 4.72–21.18) and cytogenetics (intermediate risk HR 2.49; 1.41–4.39, poor risk HR 4.74; 2.65–8.50). The score divided patients into four risk groups; good (n=47), intermediate (n=129), poor (n=198) and extremely poor (n=87). The estimated 5-y OS was 0.70 (95% CI: 0.53–0.81), 0.37 (0.28–0.46), 0.15 (0.10–0.21) and 0.03 (0.01–0.10) respectively. The model showed good discrimination with large differences between survival curves and good Harrell concordance of 0.69. It showed good calibration using Cox-Snell and deviance residuals. In the intermediate risk cytogenetic group, the model showed good discrimination with over 45% difference in 5-y OS between the good and extremely poor groups.

Conclusions:
Our study confirmed the prognostic impact of the 5 variables reported in the literature. Using these factors, we developed a score to predict long term OS that showed good discrimination and calibration. The score added further discrimination in the intermediate risk cytogenetic group. Prospective external validation of the score is needed.

Disclosures:
No relevant conflicts of interest to declare.
Overexpression of SIRT1, a Longevity Gene-Encoded Protein, and Induction of Apoptosis by Its Inhibition In Adult T-Cell Leukemia Cells
Authors: Kozako Tomohiro , Yoshimitsu Makoto , White Yohann , Aikawa Akiyoshi , Shoji Teruhisa , Honda Shinichiro , Shimeno Hiroshi , Soeda Shinji , Arima Naomichi 
Published: October 2019

Abstract
Abstract 2768
Adult T-cell leukemia-lymphoma (ATL) is an aggressive peripheral T-cell neoplasm developing after a long-term infection with human T-cell leukemia virus (HTLV-1), in which NF-kB is also implicated as an exacerbation factor. Despite recent progress in both chemotherapy and supportive care for hematological malignancies, the prognosis of ATL is still poor; overall survival at 3 years is only 24%. New strategies for the therapy and prophylaxis of ATL (e.g., vaccines and novel molecular target agents) are still required. SIRT1, an NAD+-dependent histone/protein deacetylase, plays a crucial role in various physiological processes, such as aging, metabolism, neurogenesis and apoptosis, due to its ability to deacetylate numerous substrates, such as histone and NF-kB. Here, we assessed how SIRT1 is regulated in primary ATL cells and leukemic cell lines. SIRT1 expression in ATL patients was significantly higher than that in healthy controls, especially in the acute type. Sirtinol, a SIRT1 inhibitor, induced significant growth inhibition or apoptosis in cells from ATL patients and leukemic cell lines, especially HTLV-1-related cell lines (S1T and MT-2). Sirtinol-induced apoptosis was mediated by activation of the caspase family, and inactivation of NF-kB, reducing IkBα phosphorylation. Interestingly, NAD+ augmented sirtinol-induced apoptosis following deacetylation of NF-kB via NAD+-dependent deacetylase. Thus, the SIRT1 inhibitor acted as a tumor suppressor, where NAD+ accelerated the SIRT1 inhibitor-induced apoptosis. These results suggest that SIRT1 is a crucial antiapoptotic molecule in ATL cells, and that SIRT1 inhibitors may be useful therapeutic agents for leukemia, especially in patients with ATL.
Figure 1. SIRT1 inhibitor reduces viability of leukemic cell lines. Cell lines were incubated at 1×105 cells/mL in the presence of various concentrations of sirtinol for 72 h. Proliferation of cell lines in the absence or presence of the indicated concentrations of sirtinol was assessed by WST-8 assay. Data are means ± S.D. from 3 independent experiments. Figure 1. SIRT1 inhibitor reduces viability of leukemic cell lines. Cell lines were incubated at 1×105 cells/mL in the presence of various concentrations of sirtinol for 72 h. Proliferation of cell lines in the absence or presence of the indicated concentrations of sirtinol was assessed by WST-8 assay. Data are means ± S.D. from 3 independent experiments.

Disclosures:
No relevant conflicts of interest to declare.
Short Course Adjuvant Chemotherapy for Localized Diffuse Large B-Cell Lymphoma After Complete Resection.
Authors: Jo Jungmin , Yoo Changhoon , Ahn Yongchel , Park Seong Joon , Lee Dae Ho , Kim Sang-We , Lee Jungsin , Suh Cheolwon 
Published: October 2019

Abstract
Abstract 2824

Introduction:
Diffuse large B-cell lymphoma (DLBCL) in general has been treated 6 or 8 cycles chemotherapy. Not a few patients have had adverse events during chemotherapy and stopped treatment because of treatment related toxicities. Favorable results were reported from NHL-BFM 90 trial on childhood and adolescents B-cell NHL with short intensive therapy courses depending on stage and tumor mass. Treatment was stratified into 3 risk groups and patients with initial complete resection received 2 therapy courses. We aimed to investigate effectiveness of short course chemotherapy for localized DLBCL after complete resection in adults.

Patients and Methods:
The records of Asan Medical Center registry for Non-Hodgkin's lymphoma were reviewed. Between March 2002 and March 2009, 18 patients were enrolled with DLBCL of stage I or II and complete resection before initial chemotherapy. All patients received 3 to 4 courses of rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone chemotherapy (R-CHOP) or CHOP without radiotherapy. Retrospective analysis of disease-free survival (DFS) and overall survival (OS) was performed.

Results:
The study cohort of 18 patients showed median age of 53.5 years (range 18–69 years) at diagnosis. Ten patients were males. The Ann Arbor stage was I in 14 patients, II in 4 patients. All patients of stage II had gastrointestinal lymphoma with regional lymph node involvement. Only 2 patients had B symptom. All patients were stratified into low-risk group according to international prognostic index (IPI). Most of patients (88.9%) had 3 cycles chemotherapy and 2 patients with 4 cycles. Fifteen patients were treated with R-CHOP and the others with CHOP. The median follow-up time was 27.5 months (range 6.1–100 months). The 2-year DFS was 100% and the 2-year OS was 100% in study cohort.

Conclusions:
It has been reported that 4-year OS of DLBCL was 86% in low risk group (0-1 IPI factor) treated with R-CHOP. Favorable result with short course chemotherapy showed in our study despite short follow-up time. On the result, we suggest reduced treatment for localized DLBCL after complete resection. However, prospective study is warranted to investigate effectiveness.

Disclosures:
No relevant conflicts of interest to declare.
Rituximab Plus Gemcitabine and Oxaliplatin as Salvage Therapy In Patients with Relapsed/Refractory Mantle-Cell Lymphoma.
Authors: Gironella Mercedes , López, MD Andrés , Pau Abrisqueta , Anny Jaramillo , Noelia Purroy , Bosch Francesc 
Published: October 2019

Abstract
Abstract 2825
Mantle cell lymphoma (MCL) constitutes one of the lymphomas with poorest prognosis at relapse and there are no effective salvage.
The activity shown by the combination Gemcitabine and Oxaliplatin in several types of limfoma, along with its “in vitro” synergistic effect, made this regimen an attractive regimen for salvaging patients with MCL. Against this background, we performed an off-label pilot study in order to assess efficacy and toxicity of the combination R-GemOx in relapsed or refractory patients with MCL.
For this, 27 patients (70% male, median age 70 years) diagnosed with MCL between November 2004 and January 2010 were included in this study. Inclusion criteria were adequate performance status, confirmed diagnosis of MCL and relapse or refractoriness to the previous treatment. The regimen consisted of Rituximab 375 mg/m2 on day 1, Gemcitabine 1000 mg/m2 and Oxaliplatin 100 mg/m2 on day 2, every 14 days, up to 8 cycles. Dose and interval were adjusted according to hematological and extrahematological toxicities. Median number of previous regimens was 1 (range 1 to 3), being EPOCH-R (n=14), R-CHOP (n=6), and RFC (n=3) the most frequently used induction therapies. Twelve patients relapsed after prior CR, 10 progressed after achieving a PR, whereas 5 were refractory to therapy. At inclusion, 85% of patients were in advanced (III/IV) clinical stage, 40% had bone marrow infiltration, 28% gastrointestinal involvement, and 17% cavum infiltration. Median number of cycles administered was 8 (range, 3 to 8). Doses were reduced in 9 cycles and delayed in 15 cycles. Neutropenia grade 3–4 was observed in 9 cycles and thrombocytopenia grade 3–4 in 6. Hepatotoxicity grade 1–2 in 5 pts and grade 3 in 1, sensitive neurotoxicity grade 1–2 in 12 pts, and renal impairment grade 2 in 1 patient. After completion of the treatment, 21 pts (77%) were considered in CR/uCR, 1 (4%) achieved a PR, 1 a SD, whereas 4 PD. Six patients subsequently received a stem-cell transplantation (4 allogeneic, 2 autologous). Thirteen pts are still alive and out of progression. Ten pts have died (8 due to progression and 2 due to acute GVHD). With a median follow-up of 23 months (range: 3–57), PFS and OS at 2 years are 41% and 58%, respectively. The R-GemOx combination showed a significant activity in relapsed or refractory pts with MCL with a very acceptable toxicity profile. These results prompted us to conduct a multicenter phase II clinical trial that is now ongoing.

Disclosures:
López: Roche Farma: Research Funding, Travel Support. Bosch:Roche Farma: Research Funding.
Lenalidomide In Patients with Relapsed or Refractory Hodgkin Lymphoma.
Authors: Böll Boris , Fuchs Michael , Reiners Katrin S. , Engert Andreas , Borchmann Peter 
Published: October 2019

Abstract
Abstract 2828
The immunomodulatory compound lenalidomide induces apoptosis in tumor cells, exerts antiangiogenic effects and activates immune effector cells. Apoptosis resistance, increased neoangiogenesis and impaired immunity critically contribute to Hodgkin‘s Lymphoma (HL). However, the activity of lenalidomide in HL is unknown so far.
42 patients with multiple relapsed or refractory HL were treated with 25 mg oral lenalidomide for 21 days of a 28-day cycle in a named patient program in 18 different german hospitals. Participants gave written informed consent, had no curative treatment options, an ECOG performance status ≤ 2, and normal organ functions including peripheral blood counts within the normal range. Patients were staged and restaged with computed tomography (CT) of the initially involved sites; the use of PET was optional. Response was defined according to the “Cheson-criteria” from 1999 and evaluated after two to three cycles and every other cycle thereafter. Treatment was continued until disease progression or in case of intolerable toxicity occurred. Concomitant anti-thrombotic prophylaxis with 100 mg daily oral acetylsalicylic acid was mandatory. Patients with a history of thrombosis or thrombembolic events received low molecular heparin.
To date, in the 24 patients who are eligible for analysis, treatment was well tolerated. No hematological or other toxicities above CTC grade-2 were reported; adverse events included diarrhea, constipation, neuropathy, and mild dyspnea. At restaging, eight of the analyzed patients had stable disease and twelve patients achieved clinical responses (11× PR, 1× CR) as measured by CT-scan. However, five of the heavily pretreated patients had progressive disease. Results on the remaining patients are currently being analyzed and will be presented at the ASH meeting.
In conclusion, these first clinical observations indicate that lenalidomide is well tolerated and effective in patients with relapsed or refractory HL. Consequently, international clinical trials evaluating the role of lenalidomide in HL are in preparation and a phase I/II trial evaluating lenalidomide with AVD (Adriamycine, Vinblastine and Dacarbacine) in elderly HL patients is currently recruiting.

Disclosures:
Off Label Use: Lenalidomide approved for Multiple Myeloma and MDS (5q−).